專利名稱：利用3－氨基－5－羥基苯甲酸(ahba)合酶基因保守序列篩選安莎類化合物的方法
技術(shù)領(lǐng)域：
本發(fā)明涉及一種利用參與微生物產(chǎn)物生物合成酶編碼基因保守序列從微生物中鑒定尋找新的活性物質(zhì)的方法。
背景技術(shù)：
微生物來源的產(chǎn)物如抗生素在人類與疾病的斗爭中發(fā)揮了巨大的作用。以活性為指標是目前大多數(shù)常用的篩選微生物藥物的方法，其缺點是從活性篩選出發(fā)，無法顯示化學結(jié)構(gòu)的特點，從活性篩選到化學結(jié)構(gòu)的確定，需要經(jīng)過發(fā)酵、提取分離、純化及結(jié)構(gòu)鑒定的繁雜過程，而且，對未知性質(zhì)的物質(zhì)確定提取及純化方法，需要探索及經(jīng)驗積累。因此，以活性出發(fā)的篩選方法存在盲目性大、工作量大、收效有限的諸多缺陷。
在大量微生物產(chǎn)物被發(fā)現(xiàn)及應用的基礎(chǔ)上，可以看出某些結(jié)構(gòu)類型的化合物，如青霉素、頭孢菌素等β-內(nèi)酰胺類化合物具有廣譜抗菌活性強、毒性低的特點；安莎類化合物的活性類型多，包括抗菌、抗分枝桿菌、抗麻瘋、抗腫瘤、免疫抑制、抗蟲等，最近又發(fā)現(xiàn)其具有廣譜抗病毒作用。因此，建立以鑒別化學結(jié)構(gòu)為主的篩選方法，可以比較有針對性地篩選目的結(jié)構(gòu)化合物。HansZahner[Schaal/Pulverer(eds)Actinomycetes Zbl.Bakt.Suppl.11，1981]曾提出用與化學結(jié)構(gòu)某些基團如還原糖、胺基酮酸等分子基團或生物鹼、含氮、含鹵素等起反應的試劑，建立篩選方法，他使用四氮唑蘭篩選產(chǎn)生了縮酮類(ketalin)化合物，但這種方法不能使用發(fā)酵液直接進行篩選，仍需經(jīng)過初步提取，并在TLC薄層板上進行純化，難以適用于高通量篩選。
發(fā)明目的微生物產(chǎn)物尤其是次級代謝產(chǎn)物抗生素分子生物學研究的進展表明，結(jié)構(gòu)類型相似的抗生素其生物合成編碼基因有共同性，某些基因序列有高度保守性。本發(fā)明的構(gòu)思在于，設(shè)計并利用特異結(jié)構(gòu)生物合成基因保守序列，克隆獲得特異結(jié)構(gòu)生物合成基因，以此為探針，與微生物等生物體基因組雜交，即可得到產(chǎn)生該特異結(jié)構(gòu)化合物的產(chǎn)生菌。利用這一原理，可以建立對化學類型有針對性的新篩選方法，并可用于高通量篩選。
AHBA(3-氨基-5-羥基苯甲酸)為安莎類化合物所擁有的共同結(jié)構(gòu)，是這類化合物生物合成的起始單位，因此，AHBA生物合成基因具有高度保守性。本發(fā)明的目的在于利用現(xiàn)今發(fā)展的分子生物學手段，根據(jù)安莎類化合物AHBA生物合成基因保守序列，通過克隆相關(guān)基因以尋找新的安莎類抗生素。這種方法準確度高，并可應用于其他類群化合物的篩選。

發(fā)明內(nèi)容
一.AHBA合酶基因的克隆吸水鏈霉菌(Streptomyces hygroscopicus)17997是本所從我國土壤中分離得到的一株放線菌(本所菌種保藏中心保存)，并鑒定它為已知抗生素格爾德霉素[J Antibiotics1970，23442-447]產(chǎn)生菌。格爾德霉素屬于苯醌型安莎霉素，本所發(fā)現(xiàn)格爾德霉素除了具有已知的抗腫瘤、抗細菌、抗原蟲活性，還具有廣譜抗病毒作用(陶佩珍等，1997，1998，2001)。
利用生物學軟件Vector NTIsuit6.0程序，對NCBI數(shù)據(jù)庫中所有登錄的含AHBA結(jié)構(gòu)的抗生素利福霉素B，ansatrieninA，napthomycin，絲裂霉素C，ansamitocin等的AHBA合酶編碼序列分析后，根據(jù)其氨基酸序列的保守區(qū)設(shè)計了一對簡并引物上游引物5′- AGAGGATCCTTCGAGCRSGAGTTCGC -3′BamHI下游引物5′- GCAGGATCCGGAMCATSGCCATGTAG -3′以S.hygroscopicus17997基因組DNA為模板，在LATaq酶作用下進行PCR反應，結(jié)果擴增出755bp特異性條帶(圖1)。
755bpPCR產(chǎn)物測序如下ttcgagcagg agttcgcgga cttccacggc gccccacacg ctttggccgt caccaacggc 60acccacgccc tggagttggc gttgcagtgt ctgggcgtcg ggccgggcac cgaggtcatc 120gtgccggcct tcaccttcat ctcctcctcc caggccgctc agcggctggg agcggttgcc 180gtccccgtcg acgtcgatct cgatacctac aacatcgacg tggctgccgc ggcttccgcc 240gtcacccccc tcaccaaggc gatcatgcct gtgcacatgg cggggctcat cgccgacatg 300gacgcgctcg gcgaactctc cgccgacacc ggtgtgcctc ttctccagga cgccgcccac 360gcacacggtg cccgctggca gggcaaacgg gtgggcgagt tgggtacggt cgcctcgttc 420agcttccaga acggcaagct gatgaccgcc ggcgagggcg gtgcgctgct cctgcccgac 480gaggagacct acgaggccgc gttcctgcgg cacagttgtg gccggtcacg taccgaccgc 540cgatacatgc accagaccgc cggcacgaac atgcggctca acgagttctc cgcggccgtg 600ctccgcgccc agctgggccg cctcgacgcc cagatcacgc tccgcgatca gcgctggacg 660ctgctgtccc ggctgctcgg tgagatcgac agcgtcgtac cccagggcag cgacccgcgc 720gccgaccgga actcccacta catggccatg gtccg 755分析結(jié)果顯示，該PCR產(chǎn)物編碼的氨基酸與已知的利福霉素產(chǎn)生菌Amycolatopsismediterranei中的AHBA合酶、ansatrienin、napthomycin產(chǎn)生菌S.collinus和ansamitocin產(chǎn)生菌Actinosynnema pretiosum的AHBA合酶保守區(qū)高度同源。同源性比較如下 二.吸水鏈霉菌17997中AHBA合酶基因簇的確定以上序列分析結(jié)果表明，755bpPCR產(chǎn)物含有AHBA合酶的部分編碼基因。以755bpPCR產(chǎn)物為探針，選擇高嚴謹度的雜交條件與S.hygroscopicus17997柯斯質(zhì)?；蛭膸爝M行菌落雜交及Southern雜交，最終從文庫中篩選獲得的陽性克隆可分為兩組一組柯斯質(zhì)粒是在BamHI3.0kb片段處有雜交信號；另一組柯斯質(zhì)粒是BamHI4.0kb片段處有雜交信號(圖2)。BamHI酶切分析顯示，每一組的柯斯質(zhì)粒DNA間有很好的重疊性，而兩組克隆之間未顯示出有明確的重疊性。由此表明在S.hygroscopicus17997中存在兩套與AHBA生物合成相關(guān)的基因簇。從本發(fā)明所獲得的陽性克隆酶切及Southern分析來看，這兩個生物合成基因簇在染色體上可能至少相距30kb左右。
將兩組中代表性的柯斯質(zhì)粒pCGBA10及pCGBA3進行大規(guī)模測序，測序結(jié)果以FramPlot程序分析讀碼框，通過Blast程序與NCBI數(shù)據(jù)庫中序列進行同源性比較及保守序列分析，結(jié)果發(fā)現(xiàn)除一組基因簇(pCGBA10)負責已知的格爾德霉素的生物合成以外，還發(fā)現(xiàn)并鑒定了另一組基因簇(pCGBA3)負責目前尚未發(fā)現(xiàn)的萘醌型安莎類抗生素的生物合成。三.吸水鏈霉菌17997中萘醌型安莎霉素類化合物生物合成基因的分析pCGBA3柯斯質(zhì)粒中外源片段序列分析表明，其中含有10個開放閱讀框架(ORF)，其轉(zhuǎn)錄方向及排列方式見(圖3)。它們分別編碼與AHBA生物合成相關(guān)酶AHBA合酶、磷酸化酶、氧化還原酶、氨基脫氫奎尼酸合成酶及與安莎霉素合成相關(guān)酶I型聚酮合酶和酰胺合酶。同源性分析表明，它們與萘醌型安莎霉素類生物合成酶基因更為接近。1、shnA I型聚酮合酶基因Genbank收錄號AF506522，由1452bpDNA序列組成，編碼484個氨基酸序列g(shù)tgggcgtac tggcggatgt gcacgcatcc ggcgagccgc aggtcgcact ccgatcgggc 60Val Gly Val Leu Ala Asp Val His Ala Ser Gly Glu Pro Gln Val Ala Leu Arg Ser Glygcggtcctcg tcccgcgtct cgctcgtgtc gccgatacgg accgggcttc caccggccgt 120Ala Val Leu Val Pro Arg Leu Ala Arg Val Ala Asp Thr Asp Arg Ala Ser Thr Gly Argcgtctcgacc ccgatggcac cgcgctgata accggtggta ccggtgcgct cggtgcgctg 180Arg Leu Asp Pro Asp Gly Thr Ala Leu Ile Thr Gly Gly Thr Gly Ala Leu Gly Ala Leugtggcccggc atctggtcgt cgagcacaag atccggagtc tggtcctggt aagccgtcgg 240Val Ala Arg His Leu Val Val Glu His Lys Ile Arg Ser Leu Val Leu Val Ser Arg Argggcccggacg ccccgggggc cgccgatctc gacgcggagc tgactgccct gggtgcccgc 300Gly Pro Asp Ala Pro Gly Ala Ala Asp Leu Asp Ala Glu Leu Thr Ala Leu Gly Ala Arggtgcgaattg tcgcctgcga catcgcggac cgcgaggcgg ccggggaact gatcgcctct 360Val Arg Ile Val Ala Cys Asp Ile Ala Asp Arg Glu Ala Ala Gly Glu Leu Ile Ala Sergtaccgcggg acgcgccgct cactgccgtg gtgcacacgg ccggtgtgct ggacggcggc 420Val Pro Arg Asp Ala Pro Leu Thr Ala Val Val His Thr Ala Gly Val Leu Asp Gly Glygtggtcactg cccttacgcc ggagcggctc gatgccgttc tccggccgaa ggcggacgcc 480Val Val Thr Ala Leu Thr Pro Glu Arg Leu Asp Ala Val Leu Arg Pro Lys Ala Asp Alagctctggtcc tggacgagct gacccgccac ctggacgtgg cggccttcgt cctgttctcc 540Ala Leu Val Leu Asp Glu Leu Thr Arg His Leu Asp Val Ala Ala Phe Val Leu Phe Sertccgccgccg gaacgttcgg caaccccggc cagggaaacc tggccgcctc gaacgcgtat 600Ser Ala Ala Gly Thr Phe Gly Asn Pro Gly Gln Gly Asn Leu Ala Ala Ser Asn Ala Tyrcttgacgcgc tggcggtacg acgccggact gccgggctgc ccgccacatc tgttgcctgg 660Leu Asp Ala Leu Ala Val Arg Arg Arg Thr Ala Gly Leu Pro Ala Thr Ser Val Ala Trpggggtgtggg accagaccgg catcagcggg gaccttggcg tggccgatca gcggaggatg 720Gly Val Trp Asp Gln Thr Gly Ile Ser Gly Asp Leu Gly Val Ala Asp Gln Arg Arg Metgcccggtggg gcctggcggc acactccgcc caggagggtc tggagctgtt cgacgcggcg 780Ala Arg Trp Gly Leu Ala Ala His Ser Ala Gln Glu Gly Leu Glu Leu Phe Asp Ala Alattgcgggcag akgacgcggt gctcgtggcc gcgamsmtga acttcgccgg gctgcgcgcc 840Leu Arg Ala XXX Asp Ala Val Leu Val Ala Ala XXX XXX Asn Phe Ala Gly Leu Arg Alacaggccgcct ccgaacccgt acacgtactg ctgcggggcc tcgtgcgggc cggccgtcgc 900Gln Ala Ala Ser Glu Pro Val His Val Leu Leu Arg Gly Leu Val Arg Ala Gly Arg Arggccgctcagc aggcatcctc ccgcgagggc ggcctcgccg gacagctggc tgccacgccc 960Ala Ala Gln Gln Ala Ser Ser Arg Glu Gly Gly Leu Ala Gly Gln Leu Ala Ala Thr Proccggttcagc gggagcagat cctgctggat ctggtgcggc gcgaggtcgc tgcggtcctc 1020Pro Val Gln Arg Glu Gln Ile Leu Leu Asp Leu Val Arg Arg Glu Val Ala Ala Val Leuggctattcga caccacgcaa ggtcgacccc gaccgggcct tccaggacgt cgggttcacc l080Gly Tyr Ser Thr Pro Arg Lys Val Asp Pro Asp Arg Ala Phe Gln Asp Val Gly Phe Thrtcggtgctgg ccgtcgaact ccgtaaccga ctcgccgggc tcgcggggat ccggctcccg 1140Ser Val Leu Ala Val Glu Leu Arg Asn Arg Leu Ala Gly Leu Ala Gly Ile Arg Leu Progcatcgatcg ccttcgacca tcccacaccg cggcgcatga tgcgccatct gctcgcggaa 1200Ala Ser Ile Ala Phe Asp His Pro Thr Pro Arg Arg Met Met Arg His Leu Leu Ala Gluctgtgccccg aggatggcag cgagccggcg gaccgggagg atgagatccg cagggccctg 1260Leu Cys Pro Glu Asp Gly Ser Glu Pro Ala Asp Arg Glu Asp Glu Ile Arg Arg Ala Leugcgaccacac ctctgtcccg gttccgcgag cttgggctca tggagcagat cctgcacctg 1320Ala Thr Thr Pro Leu Ser Arg Phe Arg Glu Leu Gly Leu Met Glu Gln Ile Leu His Leugtggcgcatc ccagtggtga aagtgccgca gcaccggaca ccgcggaacc gaaacaggac 1380Val Ala His Pro Ser Gly Glu Ser Ala Ala Ala Pro Asp Thr Ala Glu Pro Lys Gln Aspgccgggccgc tgatcgcgga gatggacatc gacaaccttg tgaagcgggc gatggaaaag 1440Ala Gly Pro Leu Ile Ala Glu Met Asp Ile Asp Asn Leu Val Lys Arg Ala Met Glu Lysgcccggaagc cg 1455Ala Arg Lys Pro與已知的Streptomyces avermitilis I型聚酮合酶PKS AVES2高度同源，一致性為46％，相似性為59％。
氨基酸序列分析表明，含2個保守的結(jié)構(gòu)域(圖4)，93-205位氨基酸短鏈脫氫酶；330-397位氨基酸磷酸泛酰巰基乙胺的附著位點。2、shnN酰胺合酶基因Genbank收錄號AF506519，由831bp DNA序列組成，編碼277個氨基酸序列。gtgcggagct ggaccagagc caatgtaaac gttgtttcct tctcggatga agtggagttc60Val Arg Ser Trp Thr Arg Ala Asn Val Asn Val Val Ser Phe Ser Asp Glu Val Glu PheGgcgtgttcg tcaaaaggga ctatcttcag cgcataggtt acgaaggatc cggtattccg120Gly Val Phe Val Lys Arg Asp Tyr Leu Gln Arg Ile Gly Tyr Glu Gly Ser Gly Ile Proaaccttcaga ccctggcgga acttcagtgg ctgcatctct gtagcctccc ctacgacacc180Asn Leu Gln Thr Leu Ala Glu Leu Gln Trp Leu His Leu Cys Ser Leu Pro Tyr Asp Thrggttacattc tgcatcagcc gtacgaggat tttgacatgc cccgcgtatt cgaagcggtg240Gly Tyr Ile Leu His Gln Pro Tyr Glu Asp Phe Asp Met Pro Arg Val Phe Glu Ala Valatgaaacggg gcggagtctg tttcgagttg aatttcctctt ccatcgtctc cttgtcgag300Met Lys Arg Gly Gly Val Cys Phe Glu Leu Asn Phe Leu Phe His Arg Leu Leu Val Gluatgggattcg atgcgcatgt gaattccgcc agcacggctct ccccggcgg ccagtggggt360Met Gly Phe Asp Ala His Val Asn Ser Ala Ser Thr Ala Leu Pro Gly Gly Gln Trp Glytccgagatcg agcacatggc tatccgtgtc cgtatagacga cgtggattgg ctcgtcgac420Ser Glu Ile Glu His Met Ala Ile Arg Val Arg Ile Asp Asp Val Asp Trp Leu Val Aspgtcgggcac ggaagcgtggc catcacggag cccctgcgtat cgatgaacag gcgggaagc480Val Gly His Gly Ser Val Ala Ile Thr Glu Pro Leu Arg Ile Asp Glu Gln Ala Gly Sergtggttcag atgggcacgga gttccgcttg gccacgcgggg cgagtggcgc gtccttcaa540Val Val Gln Met Gly Thr Glu Phe Arg Leu Ala Thr Arg Gly Glu Trp Arg Val Leu Glntacaagcca aagggcaggg attggcgtgac gcatatcggat gaaaatcaaa gatcgtgcc600Tyr Lys Pro Lys Gly Arg Asp Trp Arg Asp Ala Tyr Arg Met Lys Ile Lys Asp Arg Alaatttccgatt ggaatacatg gcgagaagaa ctgccgcccga cgcggaccct gtggtgccg660Ile Ser Asp Trp Asn Thr Trp Arg Glu Glu Leu Pro Pro Asp Ala Asp Pro Val Val Procggaagcggc gacgcggtgt ggagaacggg caggtgaccct cgtcgccaat ctcttcagg720Arg Lys Arg Arg Arg Gly Val Glu Asn Gly Gln Val Thr Leu Val Ala Asn Leu Phe Argtccatcatcg ggggcgagga gacggtgaag cacgtacgtg atgaagcaga gctgatcgag780Ser Ile Ile Gly Gly Glu Glu Thr Val Lys His Val Arg Asp Glu Ala Glu Leu Ile Gluatcatgacta cttactgggg agagtccgca cctatcgtcg ggtacgaacg a 831Ile Met Thr Thr Tyr Trp Gly Glu Ser Ala Pro Ile Val Gly Tyr Glu Arg與已知的利福霉素的酰胺合酶同源一致性34％，相似性47％。 該酶的功能是將聚酮鏈的羧基末端從聚酮合酶PKS轉(zhuǎn)移到芳香環(huán)的氨基形成酰胺鍵。3、shnS AHBA合酶基因Genbank收錄號AY077756，由1281bpDNA序列組成，編碼427個氨基酸序列。atgtggggcg gtaagagcgg tgcctaccgt gaacgtcgca catccggcca tgtgccccgc 60Met Trp Gly Gly Lys Ser Gly Ala Tyr Arg Glu Arg Arg Thr Ser Gly His Val Pro Argcccgctctat gccgtccgtc ggccatcggc cgagaaatat tcgaaatgaa gatttggaga 120Pro Ala Leu Cys Arg Pro Ser Ala Ile Gly Arg Glu Ile Phe Glu Met Lys Ile Trp Argcgcatgaacg cgcgacggac accagagttc cccacctggc cgcagtacga cgacggcgag 180Arg Met Asn Ala Arg Arg Thr Pro Glu Phe Pro Thr Trp Pro Gln Tyr Asp Asp Gly Glucgcaccggcc tgatccgggc cctggagcag ggccagtggt ggcgcatggg aggctcggag 240Arg Thr Gly Leu Ile Arg Ala Leu Glu Gln Gly Gln Trp Trp Arg Met Gly Gly Ser Glugtggactcct tcgagggtga gttcgcggac ttccacggcg ccccacacgc tttggccgtc 300Val Asp Ser Phe Glu Gly Glu Phe Ala Asp Phe His Gly Ala Pro His Ala Leu Ala Valaccaacggca cccacgccct ggagttggcg ttgcagtgtc tgggcgtcgg gccgggcacc 360Thr Asn Gly Thr His Ala Leu Glu Leu Ala Leu Gln Cys Leu Gly Val Gly Pro Gly Thrgaggtcatcg tgccggcctt caccttcatc tcctcctccc aggccgctca gcggctggga 420Glu Val Ile Val Pro Al Phe Thr Phe Ile Ser Ser Ser Gln Ala Ala Gln Arg Leu Glygcggttgcc gtccccgtcga cgtcgatctc gatacctaca acatcgacgt ggctgccgcg 480Ala Val Ala Val Pro Val Asp Val Asp Leu Asp Thr Tyr Asn Ile Asp Val Ala Ala Alagcttccgccg tcacccccct caccaaggcg atcatgcctg tgcacatggc ggggctcatc 540Ala Ser Ala Val Thr Pro Leu Thr Lys Ala Ile Met Pro Val His Met Ala Gly Leu Ilegccgacatgg acgcgctcgg cgaactctcc gccgacaccg gtgtgcctct tctccaggac 600Ala Asp Met Asp Ala Leu Gly Glu Leu Ser Ala Asp Thr Gly Val Pro Leu Leu Gln Aspgccgcccacg cacacggtgc ccgctggcag ggcaaacggg tgggcgagtt gggtacggtc 660Ala Ala His Ala His Gly Ala Arg Trp Gln Gly Lys Arg Val Gly Glu Leu Gly Thr Valgcctcgttca gcttccagaa cggcaagctg atgaccgccg gcgagggcgg tgcgctgctc 720Ala Ser Phe Ser Phe Gln Asn Gly Lys Leu Met Thr Ala Gly Glu Gly Gly Ala Leu Leuctgcccgacg aggagaccta cgaggccgcg ttcctgcggca cagttgtgg ccggtcacgt 780Leu Pro Asp Glu Glu Thr Tyr Glu Ala Ala Phe Leu Arg His Ser Cys Gly Arg Ser Argaccgaccgcc gatacatgca ccagaccgcc ggcacgaacat gcggctcaa cgagttctcc 840Thr Asp Arg Arg Tyr Met His Gln Thr Ala Gly Thr Asn Met Arg Leu Asn Glu Phe Sergcggccgtgc tccgcgccca gctgggccgc ctcgacgccca gatcacgct ccgcgatcag 900Ala Ala Val Leu Arg Ala Gln Leu Gly Arg Leu Asp Ala Gln Ile Thr Leu Arg Asp Glncgctggacgc tgctgtcccg gctgctcggt gagatcgacgg cgtcgtaccc cagggcagc 960Arg Trp Thr Leu Leu Ser Arg Leu Leu Gly Glu Ile Asp Gly Val Val Pro Gln Gly Sergacccgcgcg ccgaccggaa ctcccactac atggcgatgt tccggatccc cggcatatcc 1020Asp Pro Arg Ala Asp Arg Asn Ser His Tyr Met Ala Met Phe Arg Ile Pro Gly Ile Sergaggaggccc gcaacgccctc gtcgacacgc tcgtcgaggc cggcctgccc gccttcgcc 1080Glu Glu Ala Arg Asn Ala Leu Val Asp Thr Leu Val Glu Ala Gly Leu Pro Ala Phe Alagccttccggg cgatctaccg caccgacgcg ttctgggaga cggccgcgcc cgacaccacc 1140Ala Phe Arg Ala Ile Tyr Arg Thr Asp Ala Phe Trp Glu Thr Ala Ala Pro Asp Thr Thrgtcgacaagc tcgccgaaag ctgcccgcac accgaggcga tcagcaccga ctgcatctgg 1200Val Asp Lys Leu Ala Glu Ser Cys Pro His Thr Glu Ala Ile Ser Thr Asp Cys Ile Trpctgcaccatc gagtgctgct cgcctcggag gaggccctcc acaccacagc cgagatcatc 1260Leu His His Arg Val Leu Leu Ala Ser Glu Glu Ala Leu His Thr Thr Ala Glu Ile Ilegccgacgccg tggccgcacg g 1281Ala Asp Ala Val Ala Ala Arg該酶在安莎類抗生素中保守性較高，從同源性上可以看出，ShnS與萘醌型安莎類抗生素的AHBA合酶(利福霉素RifK、napthomycinNapF)同源性高于苯醌型安莎類抗生素ansatrienin的AHBA合酶(AsnF)及絲裂霉素C MitA。
RifK一致性80％，相似性86％NapF一致性74％，相似性80％AsnF一致性70％，相似性80％MitA一致性64％，相似性74％ ansmansamitocin此外，現(xiàn)在已知的AHBA合酶AsnF由386氨基酸組成，RifK、NapF、MitA及ansamitocin Asm24都由388個氨基酸組成，而ShnS卻由427個氨基酸組成，同時對本發(fā)明獲得的該基因的氨基酸序列進行保守結(jié)構(gòu)域的同源分析，結(jié)果發(fā)現(xiàn)含有3個保守的結(jié)構(gòu)域(圖5)55-419位氨基酸信號轉(zhuǎn)導系統(tǒng)中的傳感蛋白；75-171位氨基酸半胱氨酸/甲硫氨酸磷酸吡哆醛(PLP)依賴酶；98-225位氨基酸氨基轉(zhuǎn)移酶家族；ansatrienin和絲裂霉素C的AHBA合酶含有兩個結(jié)構(gòu)域，而利福霉素、naphthomycin、ansamitocin以及格爾德霉素的AHBA合酶僅含有一個與信號轉(zhuǎn)導系統(tǒng)有關(guān)的結(jié)構(gòu)域，說明shnS基因功能與已知的不同，且更為多樣。4、shnP磷酸化酶基因Genbank收錄號AF506520，由696bpDNA序列組成，編碼232個氨基酸。atgaccagag ggccggaatt ccatgcggca ccgaccatcg aacagcgccc cccggtcccc 60Met Thr Arg Gly Pro Glu Phe His Ala Ala Pro Thr Ile Glu Gln Arg Pro Pro Val Proagacacgctg tcatcttcga tctcgacggg gtcgtcgtcg acagcttcga ggtgatgggt 120Arg His Ala Val Ile Phe Asp Leu Asp Gly Val Val Val Asp Ser Phe Glu Val Met Glygaggcgttct ccctggcgta cgccgaggtc gtcggcaccg gcgaggcacc tttcgaggag 180Glu Ala Phe Ser Leu Ala Tyr Ala Glu Val Val Gly Thr Gly Glu Ala Pro Phe Glu Glutaccggcgcc accagggccg ctactttccc gacatcatgc ggatcatggg ccttccgctg 240Tyr Arg Arg His Gln Gly Arg Tyr Phe Pro Asp Ile Met Arg Ile Met Gly Leu Pro Leugagatggaag agcccttcgt ccgcgagagc taccggctcg ccgaccgcgt ccaggtgtac 300Glu Met Glu Glu Pro Phe Val Arg Glu Ser Tyr Arg Leu Ala Asp Arg Val Gln Val Tyrgacggtgtcg tcgacgtcct gcggacgctg aacgaacgcg gcctccggct ggccatcgcc 360Asp Gly Val Val Asp Val Leu Arg Thr Leu Asn Glu Arg Gly Leu Arg Leu Ala Ile Alaaccggcaagg caggcgagcg cgcccggtcc ctgctcgatg tcctcggcct gctcccgtac 420Thr Gly Lys Ala Gly Glu Arg Ala Arg Ser Leu Leu Asp Val Leu Gly Leu Leu Pro Tyrttcgcccacg tcatcggctc cgacgaggtg ccccggccca agcccgcccc tgacatcatc 480Phe Ala His Val Ile Gly Ser Asp Glu Val Pro Arg Pro Lys Pro Ala Pro Asp Ile Ileagacgcgcac tcgaactcct cgaggttccg gcggagcggg ccatcatgat cggcgacgcc 540Arg Arg Ala Leu Glu Leu Leu Glu Val Pro Ala Glu Arg Ala Ile Met Ile Gly Asp Alacccaccgacc tggccagcg cccacggcgc cgacgtcacc gccgtagccgc gctgtggggc 600Pro Thr Asp Leu Ala Ser Ala His Gly Ala Asp Val Thr Ala Val Ala Ala Leu Trp Glytgccaggaag gggccgaact gctcgccgcc gaccccgatg tcgtcctgcg gtggcccgcc 660Cys Gln Glu Gly Ala Glu Leu Leu Ala Ala Asp Pro Asp Val Val Leu Arg Trp Pro AlaGacctgctcg ccctctgccc ggccctgcc cggccac 696Asp Leu Leu Ala Leu Cys Pro Ala Leu Pro Gly His與已知的萘醌型安莎霉素napthomycin和利福霉素的AHBA生物合成相關(guān)的磷酸酶NapH、RifM同源性較高；而與ansatrienin AnsH同源性稍低NapH一致性78％，相似性87％RifM一致性77％，相似性83％AnsH一致性66％，相似性76％。
5、shnB I型聚酮合酶基因Genbank收錄號AF506523，由5316bpDNA序列組成，編碼1772個氨基酸序列。gtggacgcac gccagaacac ggagatagca gtggccagtt cagagagcaa ggtcgtcgaa 60Val Asp Ala Arg Gln Asn Thr Glu Ile Ala Val Ala Ser Ser Glu Ser Lys Val Val Glugcactgcgcg cttcactgat ggagaacgaa cggctggaga gcgaggtcca gagcatccgc 120Ala Leu Arg Ala Ser Leu Met Glu Asn Glu Arg Leu Glu Ser Glu Val Gln Ser Ile Arggacagcctca ccgagccgat cgccatcgtc ggcatggcgt gccggttccc cggcggggtg 180Asp Ser Leu Thr Glu Pro Ile Ala Ile Val Gly Met Ala Cys Arg Phe Pro Gly Gly Valtcgtcgccgg aagagttgtg ggaattgatc gcggacggcc gttccgcggt cgaggcgttc240Ser Ser Pro Glu Glu Leu Trp Glu Leu Ile Ala Asp Gly Arg Ser Ala Val Glu Ala Phecccaccaacc ggggctggga cctggagaac ctgtacgacc cggacctcga ccggcccggc 300Pro Thr Asn Arg Gly Trp Asp Leu Glu Asn Leu Tyr Asp Pro Asp Leu Asp Arg Pro Glyacgacgtacg tacgggaggg cgcgttcctg cacgacgcgg gcgagttcga cgccggcttc 360Thr Thr Tyr Val Arg Glu Gly Ala Phe Leu His Asp Ala Gly Glu Phe Asp Ala Gly Phettcggcatct cccaaagcga gacgatggtc atggacccac agcagcgcct gatgctggag 420Phe Gly Ile Ser Gln Ser Glu Thr Met Val Met Asp Pro Gln Gln Arg Leu Met Leu Gluacatcttggg aggcgttcga acgggcgggc atcgacccgg ccgctatgcg tggcaagaac 480Thr Ser Trp Glu Ala Phe Glu Arg Ala GlyIle Asp Pro Ala Ala MetArg Gly Lys Asngtcggcgtgt tcgccggcat ggccgccggg caggagtacg ggaccgcttt ccacagcatc 540Val Gly Val phe Ala Gly Met Ala Ala Gly Gln Glu Tyr Gly Thr Ala Phe His Ser Ilecccgacgagc tcgagggcta tgtgatgacc ggcggtctgg cgagcgtcct ttcgggacgg 600Pro Asp Glu Leu Glu Gly Tyr Val Met Thr Gly Gly Leu Ala Ser Val Leu Ser Gly Arggtctcctata cgttcggatt cgaggggccg gcagtcacga tcgacaccgc ctgctcctcg 660Val Ser Tyr Thr Phe Gly Phe Glu Gly Pro Ala Val Thr Ile Asp Thr Ala Cys Ser Sertccctggtgg ccctgcacat ggcagcgcag tccctgcgtt cgggcgagtc gtcgctggcg 720Ser Leu Val Ala Leu His Met Ala Ala Gln Ser Leu Arg Ser Gly Glu Ser Ser Leu Alactggtcggag gcaccaacgt gatggccacg cccactgcct tcgtgctgac cgcgcgtgca 780Leu Val Gly Gly Thr Asn Val Met Ala Thr Pro Thr Ala Phe Val Leu Thr Ala Arg Alagggggcctgg cgaaggacgg ccggtgcaag gcgttcgcgg catccgcgga tggcacgaac 840Gly Gly Leu Ala Lys Asp Gly Arg Cys Lys Ala Phe Ala Ala Ser Ala Asp Gly Thr Asntgggccgagg gcgtgggcgt cctgctgctg gagcggcttt ccgacgccgt ccgcaacggc 900Trp Ala Glu Gly Val Gly Val Leu Leu Leu Glu Arg Leu Ser Asp Ala Val Arg Asn Glycgtgaggtcc tcggcgtcgt acgggccacc gcggtgaatc aggatggcgc gtccaacgga 960Arg Glu Val Leu Gly Val Val Arg Ala Thr Ala Val Asn Gln Asp Gly Ala Ser Asn Glyctcgccgcgc ccaacgggcc ctcgcagcag cgggtgatcc gccaggcact ggcggccggc 1020Leu Ala Ala Pro Asn Gly Pro Ser Gln Gln Arg Val Ile Arg Gln Ala Leu Ala Ala Glyggcctgtcgc cggccgacgt cgacatcgtc gaggcgcacg gcaccggaac cgccctcggc 1080Gly Leu Ser Pro Ala Asp Val Asp Ile Val Glu Ala His Gly Thr Gly Thr Ala Leu Glygaccccatcg aggcacaggc gctcctcacc acctatggtc ggaaccgtgc ccccggactg 1140Asp Pro Ile Glu Ala Gln Ala Leu Leu Thr Thr Tyr Gly Arg Asn Arg Ala Pro Gly Leuccgctgtggc ttggttcggt gaagtcgaac cttggacacg cgggcgccgc tgcgggcgtc 1200Pro Leu Trp Leu Gly Ser Val Lys Ser Asn Leu Gly His Ala Gly Ala Ala Ala Gly Valgccggtgtga tcaagatggt gatggcgatg cggcatggtg tgctgccgcg gacactgcat 1260Ala Gly Val Ile Lys Met Val Met Ala Met Arg His Gly Val Leu Pro Arg Thr Leu Hisgtggacgagc cgacgcccga ggtcgactgg tctgccggag cggtcgagct gctgaccgag 1320Val Asp Glu Pro Thr Pro Glu Val Asp Trp Ser Ala Gly Ala Val Glu Leu Leu Thr Glugcgcacgagt ggcccgaggt cggccgtcct cgccgtgcgg gggtctccgg cttcggcgcc 1380Ala His Glu Trp Pro Glu Val Gly Arg Pro Arg Arg Ala Gly Val Ser Gly Phe Gly Alaagcggcacca acgcacacgt catcctggag caggcgaccg agccgacatc cgggaacctg 1440Ser Gly Thr Asn Ala His Val Ile Leu Glu Gln Ala Thr Glu Pro Thr Ser Gly Asn Leucccgacgaga aggcacgcgt gctgggcgac tcggttgtgg cggtggtcgt ctcggcccgt 1500Pro Asp Glu Lys Ala Arg Val Leu Gly Asp Ser Val Val Pro Leu Val Val Ser Ala Argggcaaggcgg gtcttgccgg ccaggctcac cgtctcggct cgttcctgac acagcgtcaa 1560Gly Lys Ala Gly Leu Ala Gly Gln Ala His Arg Leu Gly Ser Phe Leu Thr Gln Arg Glngacacggacg tgctcgacat cggccagtcg ctggtgcgga gccggggtcc actccaggac 1620Asp Thr Asp Val Leu Asp Ile Gly Gln Ser Leu Val Arg Ser Arg Gly Pro Leu Gln Aspcgtgcggtcg tgctcgccgc ggaccgggac gaggcgctgg ccggactcga cgctgtggcc 1680Arg Ala Val Val Leu Ala Ala Asp Arg Asp Glu Ala Leu Ala Gly Leu Asp Ala Val Alacgcgccgagt ccgcgcccgg tgtggtcacg ggatttgccg agagcacagt gggccggacc 1740Arg Ala Glu Ser Ala Pro Gly Val Val Thr Gly Phe Ala Glu Ser Thr Val Gly Arg Thrgtcctcgtgt tccccggcca gggcacacag tgggcgggaa tgggagcgga actgctcgaa 1800Val Leu Val Phe Pro Gly Gln Gly Thr Gln Trp Ala Gly Met Gly Ala Glu Leu Leu Glugcctcacctg tgttcgcagc caggatgacc gagtgcgccg aggtgctcga cccgctgacc 1860Ala Ser Pro Val Phe Ala Ala Arg Met Thr Glu Cys Ala Glu Val Leu Asp Pro Leu Thrggctggtcgc tgctcgatgt ggtacggcag gtggagggcg cccggtctct tgaagacgtc 1920Gly Trp Ser Leu Leu Asp Val Val Arg Gln Val Glu Gly Ala Arg ser Leu Glu Asp Valgacgtcttgc agccggtgtc gtgggcactg atggtgtcgc tggccgcgtt gtgggaggcg 1980Asp Val Leu Gln Pro Val Ser Trp Ala Leu Met Val Ser Leu Ala Ala Leu Trp Glu Alatgcggggtcg tcccggacgc tgtcgtgggt cattccctgg gcgagatcgc cgctgcctgc 2040Cys Gly Val Val Pro Asp Ala Val Val Gly His Ser Leu Gly Glu Ile Ala Ala Ala Cystatgccggtg cgctgtccct tcccgacgcc gcccgcctca tggttcaccg gtccaggatt 2100Tyr Ala Gly Ala Leu Ser Leu Pro Asp Ala Ala Arg Leu Met Val His Arg Ser Arg Ilegccgaagccg agctggtggg acgcggaggc atggcgtccc tcaccgccga tgtcaaggcc 2160Ala Glu Ala Glu Leu Val Gly Arg Gly Gly Met Ala Ser Leu Thr Ala Asp Val Lys Alagtctccattc tgatcgagga gtggccgggt ctggagatcg ccgcggtcaa cggacccgcc 2220Val Ser Ile Leu Ile Glu Glu Trp Pro Gly Leu Glu Ile Ala Ala Val Asr Gly Pro Alatccgtggtgg tgaccggtga actgccctcc ctggaagagc tgctcgcccg atgcgaagcc 2280Ser Val Val Val Thr Gly Glu Leu Pro Ser Leu Glu Glu Leu Leu Ala Arg Cys Glu Alagacggcatcc gcgcccgcag gattcgcggc atcaacggcg ccgcacactc ctcacagatc 2340Asp Gly Ile Arg Ala Arg Arg Ile Arg Gly Ile Asn Gly Ala Ala His Ser Ser Gln Ilegacgtgctgc acgactcttt cgtggaggcc ctcgcctcgg tctccgccgg ggcttcgcgc 2400Asp Val Leu His Asp Ser Phe Val Glu Ala Leu Ala Ser Val Set Ala Gly Ala Ser Arggtaccgctgt actccacggt gaccgggcga ctccatgaca ccacggagtt cgacgtcgag 2460Val Pro Leu Tyr Ser Thr Val Thr Gly Arg Leu His Asp Thr Thr Glu Phe Asp Val Glucactggttcc gcaacatgcg gcagaccgtg cagttcgacc cggccatccg gtccctggtc 2520His Trp Phe Arg Asn Met Arg Gln Thr Val Gln Phe Asp Pro Ala Ile Arg Ser Leu Valggcgacgggc acggcgtgtt catcgaggtc agtgctcatc ctgtgctgac gtcgagcgtc 2580Gly Asp Gly His Gly Val Phe Ile Glu Val Ser Ala His Pro Val Leu Thr Ser Ser Valcaggacgtgc tggcggacct cgaggccgga ccggccgtcg tcaccgggac gctgcgccgc 2640Gln Asp Val Leu Ala Asp Leu Glu Ala Gly Pro Ala Val Val Thr Gly Thr Leu Arg Arggacgacggcg gcccgcgccg gttcctcgcc tcgctggccc acctgtacac ccacggcgta 2700Asp Asp Gly Gly Pro Arg Arg Phe Leu Ala Ser Leu Ala His Leu Tyr Thr His Gly Valcgggtcgact gggaagccgt cctcggtcgc ggcggggaac agcccgtaga cctgccgacg 2760Arg Val Asp Trp Glu Ala Val Leu Gly Arg Gly Gly Glu Gln Pro Val Asp Leu Pro Thrtacgcctttc agcgccagcg gtactggctg gagacggcag agtcccgtgg ggacgcaccg 2820Tyr Ala Phe Gln Arg Gln Arg Tyr Trp Leu Glu Thr Ala Glu Ser Arg Gly Asp Ala Proggcctcggtc tggaggtggc gaaccatccc ctgctcggcg cggttaccga gatccccggc 2880Gly Leu Gly Leu Glu Val Ala Asn His Pro Leu Leu Gly Ala Val Thr Glu Ile Pro Glytcggacggcg tgctgttcac ttcccggctg tcgctgcgca cacacccctg gctcgccgac 2940Ser Asp Gly Val Leu Phe Thr Ser Arg Leu Ser Leu Arg Thr His Pro Trp Leu Ala Aspcacgcgggcg ccggagtcgt cctcctgccg ggagcggcct tcgtggaact cgcagtccgt 3000His Ala Gly Ala Gly Val Val Leu Leu Pro Gly Ala Ala Phe Val Glu Leu Ala Val Arggccgcggacg aggtcggcta cgggctggtc ggcgaactgg tcatcgagcg ccccctggtg 3060Ala Ala Asp Glu Val Gly Tyr Gly Leu Val Gly Glu Leu ValIle Glu Arg Pro Leu Valctgcccgaga gcggcggcgt ccaggtacgc gtgtgggtcg gcgagcccga cgagtccggc 3120Leu Pro Glu Ser Gly Gly Val Gln Val Arg Val Trp Val Gly Glu Pro Asp Glu Ser Glycaccgtaccg tccaggttca ctcccgccgg gaggaagccg gctcgcgagg gagctggacc 3180His Arg Thr Val Gln Val His Ser Arg Arg Glu Glu Ala Gly Ser Arg Gly Ser Trp Thrcgtcatgtct ccgggcggct ggtgccggag gacggccagg ccgagttcga cctcacccag 3240Arg His Val Ser Gly Arg Leu Val Pro Glu Asp Gly Gln Ala Glu Phe Asp Leu Thr Glntggccgccgc ccggcgccac cgcggtcgac ccggacgcgt tcgcccacgc gtacgaccac 3300Trp Pro Pro Pro Gly Ala Thr Ala Val Asp Pro Asp Ala Phe Ala His Ala Tyr Asp Histtggcagagg cgggatacca ctatggtcca gcctttcagg gaatgcgcgc ggcttggact 3360Leu Ala Glu Ala Gly Tyr His Tyr Gly Pro Ala Phe Gln Gly Met Arg Ala Ala Trp Thrcgtggcgagg aggtgttcgc cgaggtctca ctgccggagt cggcgggcaa ggccgatgag 3420Arg Gly Glu Glu Val Phe Ala Glu Val Ser Leu Pro Glu Ser Ala Gly Lys Ala Asp Glutacgggttgc acccggccct gctggacgcg gccatgcaca ccagtctctt ccgccccgat 3480Tyr Gly Leu His Pro Ala Leu Leu Asp Ala Ala Met His Thr Ser Leu Phe Arg Pro Aspctgagcgacg agagcccgaa gctggctctg ccgttcgtct ggcgcgatgt caggctgcac 3540Leu Ser Asp Glu Ser Pro Lys Leu Ala Leu Pro Phe Val Trp Arc Asp Val Arg Leu Hisgccgacggag cctccacgct gcgggtgcac ctcaccccgc tcgcccccga cacgatccgc 3600Ala Asp Gly Ala Ser Thr Leu Arg Val His Leu Thr Pro Leu Ala Pro Asp Thr Ile Argttgcacctgg ccgacacttc cggcacaccc gtggcttcgg tcgactcgtt ggtcctgcgc 3660Leu His Leu Ala Asp Thr Ser Gly Thr Pro Val Ala Ser Val Asp Ser Leu Val Leu Argcccgtggtcc cggaactgct gcgcgtcggc tcaggcgcgg ccaaggacca gatgttccgg 3720Pro Val Val Pro Glu Leu Leu Arg Val Gly Ser Gly Ala Ala Lys Asp Gln Met Phe Arggtggcctggg agcccatctc cgtcaggagc gtggacgacg agctgaaggc cgtacgcgtg 3780Val Ala Trp Glu Pro Ile Ser Val Arg Ser Val Asp Asp Glu Leu Lys Ala Val Arg Valacgactgccg aggacgtccg tgccgcggcc gcaacggccc cgcgtgtgct cctgctcgat 3840Thr Thr Ala Glu Asp Val Arg Ala Ala Ala Ala Thr Ala Pro Arg Val Leu Leu Leu Aspgtggccggcg atggacgtac ggaccccgac gcggcccggg acctcagcgg gcgggtgctg 3900Val Ala Gly Asp Gly Arg Thr Asp Pro Asp Ala Ala Arg Asp Leu Ser Gly Arg Val Leugaggccgtcc aggcgtggct ggcggagccc gccttccagg acactgttct cctcgctctc 3960Glu Ala Val Gln Ala Trp Leu Ala Glu Pro Ala Phe Gln Asp Thr Val Leu Leu Ala Leuacacactccg gggcggccgt ccgggatggg gacccggttc ccgatctcgc cgttgcgacg 4020Thr His Ser Gly Ala Ala Val Arg Asp Gly Asp Pro Val Pro Asp Leu Ala Val Ala Thrgccgccggcc tgctgcgtgc ggcgcagtcc gagaacgtgg gccgcatcat cctggtcgac 4080Ala Ala Gly Leu Leu Arg Ala Ala Gln Ser Glu Asn Val Gly Arg Ile Ile Leu Val Aspacggacggca cggaggcgtc agcccggcgc ctgcccgatg tgctcgcggc cggggaaccg 4140Thr Asp Gly Thr Glu Ala Ser Ala Arg Arg Leu Pro Asp Val Leu Ala Ala Gly Glu Procaggcggcac ttcggtcggg ttcggtggcg gtgccgaggc tcgtcagggc ctcccccgcc 4200Gln Ala Ala Leu Arg Ser Gly Ser Val Ala Val Pro Arg Leu Val Arg Ala Ser Pro Alagaggcccagg gccgtccgct gaaccccggg ggtacggttc tgatcaccgg cggtacgggt 4260Glu Ala Gln Gly Arg Pro Leu Asn Pro Gly Gly Thr Val Leu Ile Thr Gly Gly Thr Glytcgctgggtc gcctggcggc cgggcacctg gtcaccgagc acaagatcag gagtctgctc4320Ser Leu Gly Arg Leu Ala Ala Gly His Leu Val Thr Glu His Lys Ile Arg Ser Leu Leuctggtgagcc ggcaaggacc ggacgcaccg ggcgcggccg agctggaggc cgaactcacg 4380Leu Val Ser Arg Gln Gly Pro Asp Ala Pro Gly Ala Ala Glu Leu Glu Ala Glu Leu Thrgaactcggcg cgaacgtccg gatcgtcgcg tgcgacgtct ccgatcggga ctccgtggcc 4440Glu Leu Gly Ala Asn Val Arg Ile Val Ala Cys Asp Val Ser Asp Arg Asp Ser Val Alagcgctgctgg cctctgttcc ccacgacgcc ccgctcaccg gcgtgatcca cgcagccggg 4500Ala Leu Leu Ala Ser Val Pro His Asp Ala Pro Leu Thr Gly Val Ile His Ala Ala Glygtgctggatg acggtgtggt cacctccctg acgcccgaac ggctcgacac ggtgctccgt 4560Val Leu Asp Asp Gly Val Val Thr Ser Leu Thr Pro Glu Arg Leu Asp Thr Val Leu Argcccaaggccg acgcggcaca gatcctggac gaactcacgc gcgatctcga ccttgccgtc 4620Pro Lys Ala Asp Ala Ala Gln Ile Leu Asp Glu Leu Thr Arg Asp Leu Asp Leu Ala Valttcgtcctgt actcctccat cgcggggatc ttcggttcag cgggccagag cagctatgcc 4680Phe Val Leu Tyr Ser Ser Ile Ala Gly Ile Phe Gly Ser Ala Gly Gln Ser Ser Tyr Alagccgcgaact cgttcctcga cgcgctcgct gaacgccgtc gcgcttgcgg actgccggcg 4740Ala Ala Asn Ser Phe Leu Asp Ala Leu Ala Glu Arg Arg Arg Ala Cys Gly Leu Pro Alaacctcactgg tgtggggatg gtggggccag gtgtccggca tagtggacaa gctcgccgag 4800Thr Ser Leu Val Trp Gly Trp Trp Gly Gln Val Ser Gly Ile Val Asp Lys Leu Ala Glugtcgacctga agcgcttcga ccggctcaac atgatcgagt tcaccgcaca agagggcatg 4860Ala Ser Pro Val Phe Ala Ala Arg Met Thr Glu Cys Ala Glu Val Leu Asp Pro Leu Thrgagctgttcg accttgcgct gtccgatcgc agcgctgcct tggtcctggc gaagatggac 4920Glu Leu Phe Asp Leu Ala Leu Ser Asp Arg Ser Ala Ala Leu Val Leu Ala Lys Met Aspctcaaagcaa tgcgggacca gaccgactcc gcatctgtcg ccccgctgct gcgcggcctc 4980Leu Lys Ala Met Arg Asp Gln Thr Asp Ser Ala Ser Val Ala Pro Leu Leu Arg Gly Leugtccgcgtgg gtcggcgggc cgccagtgac gggactrccg ggyccgycgg gctggcaggg 5040Val Arg Val Gly Arg Arg Ala Ala Ser Asp Gly Thr XXX Gly XXX XXX Gly Leu Ala Glycgyktgyccg aggcgtcckc cgaccagcgc ggaaagatct tggccgactt ggtccagcgc 5100XXX XXX XXX Glu Ala Ser XXX Asp Gln Arg Gly Lys Ile Leu Ala Asp Leu Val Gln Arggaggtctccg cgatcctcgg tcacctgtcg ccggaccaga tcggattgga cctgtccttc 5160Glu Val Ser Ala Ile Leu Gly His Leu Ser Pro Asp Gln Ile Gly Leu Asp Leu Ser Phettcgacctcg ggttcgactc gctgaccgcc gtcgagctcg cgaaccggct gtcggcgctg 5220Phe Asp Leu Gly Phe Asp Ser Leu Thr Ala Val Glu Leu Ala Asn Arg Leu Ser Ala Leuaccggcctcc gtatcccgtc caccttcgcc ttcgactgcc ccacggtgga cctggctgtc 5280Thr Gly Leu Arg Ile Pro Ser Thr Phe Ala Phe Asp Cys Pro Thr Val Asp Leu Ala Valgaggcgctgc tggagagctt cgaactcgac gtggacGlu Ala Leu Leu Glu Ser Phe Glu Leu Asp Val Asp與已知的利福霉素I型PKS一致性53％，相似性66％。
氨基酸序列分析顯示，含有7個相似的結(jié)構(gòu)域(圖6)44-296氨基酸β酮酰基合酶N端結(jié)構(gòu)域；186-243位氨基酸硫酯酶N端結(jié)構(gòu)域；304-472位氨基酸β酮?；厦窩端結(jié)構(gòu)域；580-885位氨基酸?；D(zhuǎn)移酶結(jié)構(gòu)域；1409-1571位氨基酸短鏈脫氫酶；1409-1552位氨基酸3-β羥基固醇脫氫酶/異構(gòu)酶家族；1702-1763位氨基酸磷酸泛酰巰基乙胺的附著位點。6、shnQ氨基脫氫奎尼酸合成酶基因Genbank收錄號AF506524，由1068bpDNA序列組成，編碼356個氨基酸序列。gtgagaggcc gaataccggt aagcatcggc gaccgttcgt acgaggtgct ggtcggccgg 60Val Arg Gly Arg Ile Pro Val Ser Ile Gly Asp Arg Ser Tyr Glu Val Leu Val Gly Argggggtgcgtt cctcgctggc cgaagtgatc cagggcctcg gcgcccggcg ggtcgccgtc 120Gly Val Arg Ser Ser Leu Ala Glu Val Ile Gln Gly Leu Gly Ala Arg Arg Val Ala Valgtgtcagccc ggcccgcgga atgggtgccc gacaccggcg tggagaccct gctgctgcct 180Val Ser Ala Arg Pro Ala Glu Trp Val Pro Asp Thr Gly Val Glu Thr Leu Leu Leu Progcccgcgacg gtgagcgggac aagaccctc gccaccgtcg aggcgctgtg cgcggagttc 240Ala Arg Asp Gly Glu Arg Asp Lys Thr Leu Ala Thr Val Glu Ala Leu Cys Ala Glu Phegtacgcttcg gcctcacccgc aacgacgcgg ttgtctcctg cgggggcgg aaccaccacc 300Val Arg Phe Gly Leu Thr Arg Asn Asp Ala Val Val Ser Cys Gly Gly Gly Thr Thr Thrgatgtcgtcg gcctggccgc cgccctgtac caccggggcg tgccagtggt gcatctgccg 360Asp Val Val Gly Leu Ala Ala Ala Leu Tyr His Arg Gly Val Pro Val Val His Leu Proaccacgttgc tggcgcaggt cgacgcgagc gtcggcggta agacggcggt caacctgccc 420Thr Thr Leu Leu Ala Gln Val Asp Ala Ser Val Gly Gly Lys Thr Ala Val Asn Leu Protccggtaaga acctggtgggc gcctattgg cagcctgccg ccgtattgtg cgacaccgag 480Ser Gly Lys Asn Leu Val Gly Ala Tyr Trp Gln Pro Ala Ala Val Leu Cys Asp Thr Glutacctgtcca ccctgccccg gcgcgagatg ctcaacggct tgggcgagat cgcccgctgc 540Tyr Leu Ser Thr Leu Pro Arg Arg Glu Met Leu Asn Gly Leu Gly Glu Ile Ala Arg Cyscatttcatcg gcgccggcga cctgcgcgag ctcggcctca cggagcgtat cgcggccagt 600His Phe Ile Gly Ala Gly Asp Leu Arg Glu Leu Gly Leu Thr Glu Arg Ile Ala Ala Sergtgacgctca aggccggcgt cgtctcggcc gatgagcgcg acaccggtct gcggcacatc 660Val Thr Leu Lys Ala Gly Val Val Ser Ala Asp Glu Arg Asp Thr Gly Leu Arg His Ilectcaactacg gccacaccct gggccacgcg ctggaatccg ccaccggctt tgcgcttcgg 720Leu Asn Tyr Gly His Thr Leu Gly His Ala Leu Glu Ser Ala Thr Gly Phe Ala Leu Argcacggcgagg ccgtcgctgt cggcacgatt ttcgcgggcc tgctcgccgg cgctctggac 780His Gly Glu Ala Val Ala Val Gly ThrIle Phe Ala Gly Leu Leu Ala Gly Ala Leu Aspcggatcggcc ccgggcgggt ggcggagcac cgggaggtcg tcgagttcta cggcctgccg 840Arg Ile Gly Pro Gly Arg Val Ala Glu His Arg Glu Val Val Glu Phe Tyr Gly Leu Progccgcgctgc ccgaggagat cgagacggct gagctgatcc gcctgatgcg cagggacaag 900Ala Ala Leu Pro Glu Glu Ile Glu Thr Ala Glu Leu Ile Arg Leu Met Arg Arg Asp Lysaaggcgctca ccggcctcgc gttcgtcctc gacggcccca gcggcgtgga gctggtccac 960Lys Ala Leu Thr Gly Leu Ala Phe Val Leu Asp Gly Pro Ser Gly Val Glu Leu Val Hisgacgtatccg aacaggtcgt cgccgatgtc ctggagcaca tgccacggca gccactcggc 1020Asp Val Ser Glu Gln Val Val Ala Asp Val Leu Glu His Met Pro Arg Gln Pro Leu Glycgactcgtcg agccggcaca cttccacgga acaggagatc cgctggca 1068Arg Leu Val Glu Pro Ala His Phe His Gly Thr Gly Asp Pro Leu Ala與已知的利福霉素，napthomycin氨基脫氫奎尼酸合酶(rifG，同源性高；與ansamitocin及MitomycinC的氨基脫氫奎尼酸合酶同源性略低利福霉素rifG一致性74％，相似性83％napthomycin 一致性75％，相似性82％ansamitocin 一致性70％，相似性79％絲裂霉素C 一致性69％，一致性77％ shnQ A-- 356mitP --- 343ansA --- 248napC AVT 358asam ---342rifG ---351asamansamitocin該基因的功能是使氨基-3-脫氧-D阿拉伯-庚酮糖酸-7-磷酸轉(zhuǎn)變?yōu)榘被?-脫氧-D阿拉伯-脫氫奎尼酸，為AHBA合成提供前體物質(zhì)。7、shnO氧化還原酶基因Genbank收錄號AF506521，由1098bp核苷酸序列組成，編碼366個氨基酸。gtgagggtcc gggctgccgt cgtcgggctc ggctgggcgg gccgcgacct gtggctcaga 60Val Arg Val Arg Ala Ala Val Val Gly Leu Gly Trp Ala Gly Arg Asp Leu Trp Leu Argctgctgcgcg agcacaagga cttcgaggtg gtcgccggtg tcgaccccga cccggactcc 120Leu Leu Arg Glu His Lys Asp Phe Glu Val Val Ala Gly Val Asp Pro Asp Pro Asp Sercgggcggccg ctgtggccac cggtctgcgc gcccacccca ccgtggacgc cctcgatccc 180Arg Ala Ala Ala Val Ala Thr Gly Leu Arg Ala His Pro Thr Val Asp Ala Leu Asp Procgcacggtcg acatcgccgt cgtcgcggta cccaaccatc tgcatgccga ggttgcggcc 240Arg Thr Val Asp Ile Ala Val Val Ala Val Pro Asn His Leu His Ala Glu Val Ala Alagccctgctcc gccgagggat ctccgtcttt ctggagaagc cggtctgcct caccactgcc 300Ala Leu Leu Arg Arg Gly Ile Ser Val Phe Leu Glu Lys Pro Val Cys Leu Thr Thr Alagaggccgacg ccctcgccga ggccgaaggc aacggcgcgg tgctgctcgc cggcagtgcc 360Glu Ala Asp Ala Leu Ala Glu Ala Glu Gly Asn Gly Ala Val Leu Leu Ala Gly Ser Alagccgcccacc gcggcgacat ccgcgagctc tccgggctgc ttccccagct cggccggatc 420Ala Ala His Arg Gly Asp Ile Arg Glu Leu Ser Gly Leu Leu Pro Gln Leu Gly Arg Ilecgccatgccg acctgtcctg ggtcagggcg cgcggagtgc cgcagcccgg cggctggttc 480Arg His Ala Asp Leu Ser Trp Val Arg Ala Arg Gly Val Pro Gln Pro Gly Gly Trp Pheacccagcgca gcagggccgg tggcggtgcg ctcgtcgacc tcggctggca ccttctcgac 540Thr Gln Arg Ser Arg Ala Gly Gly Gly Ala Leu Val Asp Leu Gly Trp His Leu Leu Aspgtcctcgcct tcctcctcgg ccccgccccc gtcgcccagg tgatcggctc gatctccgac 600Val Leu Ala Phe Leu Leu Gly Pro Ala Pro Val Ala Gln Val Ile Gly Ser Ile Ser Aspgacttcgtca gcagcagagc ctggtccgcc acgtggcgtg aggaccagct caccgacgcc 660Asp Phe Val Ser Ser Arg Ala Trp Ser Ala Thr Trp Arg Glu Asp Gln Leu Thr Asp Alaccgaccggcg acgtcgagga caccgcccgc ggcttcctggt ccgcgaggac ggtatctct 720Pro Thr Gly Asp Val Glu Asp Thr Ala Arg Gly Phe Leu Val Arg Glu Asp Gly Ile Sergtctcgttgc gggccagttg ggcctcacac gaggcgctgga cggctccgtc atcaccatc 780Val Ser Leu Arg Ala Ser Trp Ala Ser His Glu Ala Leu Asp Gly Ser Val Ile Thr Ilegagggcagcg atggaacggc acggctgcac tgcaccttcgg cttcagcccg aaccgggct 840Glu Gly Ser Asp Gly Thr Ala Arg Leu His Cys Thr Phe Gly Phe Ser Pro Asn Arg Alacccgaatcgg tgctcaccct cacgcaggat ggctccacaca gcggatccc gctgcccgcc 900Pro Glu Ser Val Leu Thr Leu Thr Gln Asp Gly Ser Thr Gln Arg Ile Pro Leu Pro Alagaacccatcg gcattgagta cggtcgccag ctcgacggcc tcgcccggct cctggccgac 960Glu Pro Ile Gly Ile Glu Tyr Gly Arg Gln Leu Asp Gly Leu Ala Arg Leu Leu Ala Aspccgggccgac ggggccaggc cgtcgcccag gcccgcagca ccgtccggtt gatcgagagc 1020Pro Gly Arg Arg Gly Gln Ala Val Ala Gln Ala Arg Ser Thr Val Arg Leu Ile Glu SerTtctatgcat cggcgcgcgc tgccccacct gtggatcacg cgtccgaatt caccgcccac 1080Phe Tyr Ala Ser Ala Arg Ala Ala Pro Pro Val Asp His Ala Ser Glu Phe Thr Ala Hisaaagaggtga ggatcgca 1098Lys Glu Val Arg Ile Ala與napthomycin AHBA生物合成相關(guān)的氧化還原酶NapG一致性47％，相似性198/35056％。
氨基酸序列分析顯示，shnO含有5個保守的結(jié)構(gòu)域(圖7)3-96位氨基酸和137-191位氨基酸均為氧化還原酶家族；2-87位氨基酸乙醛脫氫酶家族，細菌中該酶催化在3-羥基丙酸降解途徑中催化乙醛形成乙酰輔酶A；7-82位氨基酸預苯酸脫氫酶家族，該酶涉及酪氨酸的生物合成；2-91位氨基酸細菌中多聚糖生物合成蛋白。8、ShnC GenBank收錄號AF521897，由2322bp DNA序列組成，編碼774個氨基酸序列。atggccgagc tcttcgtccg gggtgttccc gtcgactgga ccaagttcct catggccggg 60Met Ala Glu Leu Phe Val Arg Gly Val Pro Val Asp Trp Thr Lys Phe Leu Met Ala Glygccgggcacg tcgaccttcc gacgtacgcc ttcgaccggc gccactactg gttgcaggat 120Ala Gly His Val Asp Leu Pro Thr Tyr Ala Phe Asp Arg Arg His Tyr Trp Leu Gln Aspgctgcgacag ccgacgacag cggcgcctcc gacaacgacg ccgacgcgga cttctggagc 180Ala Ala Thr Ala Asp Asp Ser Gly Ala Ser Asp Asn Asp Ala Asp Ala Asp Phe Trp Sergccgtcgagc agaccgacgc ggactcgctc gccgggctcc tcgccccgga ctccgccggt 240Ala Val Glu Gln Thr Asp Ala Asp Ser Leu Ala Gly Leu Leu Ala Pro Asp Ser Ala Glyctgcgcgacg ccttgcgcac cgtcgtgccg gcgcttgcgg actggcgcgg caggagccgg 300Leu Arg Asp Ala Leu Arg Thr Val Val Pro Ala Leu Ala Asp Trp Arg Gly Arg Ser Argcggcgctcca gcgctgaacg cctccgctac gccgtcacct ggcggcccct ggaccgtgag 360Arg Arg Ser Ser Ala Glu Arg Leu Arg Tyr Ala Val Thr Trp Arg Pro Leu Asp Arg Glugtgtcaaggg tccccgcggg ccgctggctc gccgtcctgc cgccgggatg cccggccgaa 420Val Ser Arg Val Pro Ala Gly Arg Trp Leu Ala Val Leu Pro Pro Gly Cys Pro Ala Gluaccgtgaccg gctcccgggt ggccgagctc atcgcggagc tcggtgccca gggactcgac 480Thr Val Thr Gly Ser Arg Val Ala Glu Leu Ile Ala Glu Leu Gly Ala Gln Gly Leu Aspgtggtgccct tcgagaccgt tccctccgcc ttcacccgca ccggactcac cgcgcgcctg 540Val Val Pro Phe Glu Thr Val Pro Ser Ala Phe Thr Arg Thr Gly Leu Thr Ala Arg Leuagcgacatcc gggccgagta ccagcccgcg ggagtcctct ccctgctcgc cctcgacggc 600Ser Asp Ile Arg Ala Glu Tyr Gln Pro Ala Gly Val Leu Ser Leu Leu Ala Leu Asp Glygagcaggacg tcatcgacac cgtcgccagg accctcgcgc tggttcaggc gctgggggac 660Glu Gln Asp Val Ile Asp Thr Val Ala Arg Thr Leu Ala Leu Val Gln Ala Leu Gly Aspgcgggcgtga acgggccgct gtggtgtctg acccggggcg cggtgaacac cggaattcag 720Ala Gly Val Asn Gly Pro Leu Trp Cys Leu Thr Arg Gly Ala Val Asn Thr Gly Ile Glngacacggccg gcgagcccgg cgacgccgcg atctgggggc tgggccgcgc cgcggctctc 780Asp Thr Ala Gly Glu Pro Gly Asp Ala Ala Ile Trp Gly Leu Gly Arg Ala Ala Ala Leugagcaccccg accggtgggg cggcctgatc gacctgccgg cgaccgccga cgcccacacc 840Glu His Pro Asp Arg Trp Gly Gly Leu Ile Asp Leu Pro Ala Thr Ala Asp Ala His Thrgcgcagtacc tcgtgggcgc gctgaacggc accgcgggcg accagctcgc cgtacgccgc 900Ala Gln Tyr Leu Val Gly Ala Leu Asn Gly Thr Ala Gly Asp Gln Leu Ala Val Arg ArgCccggcctct acagccggcg gctcgtacgc aagcccgcgt ctcagacccc cgccgacggc 960Pro Gly Leu Tyr Ser Arg Arg Leu Val Arg Lys Pro Ala Ser Gln Thr Pro Ala Asp Glyggctggcggc cccacggcac agtcctcgtg accggcggcg ccgaggccct cggcatccat 1020Gly Trp Arg Pro His Gly Thr Val Leu Val Thr Gly Gly Ala Glu Ala Leu Gly Ile Hisgcctcgctct ggctcgcccg gtccggcgcg cgccgtctca tcgtcacaac cacggctcag1080Ala Ser Leu Trp Leu Ala Arg Ser Gly Ala Arg Arg Leu Ile Val Thr Thr Thr Ala Glngcccccgccg acgccgtcac cgagttgcag ggcaagctcg cggccgccgg ggtggagacg 1140Ala Pro Ala Asp Ala Val Thr Glu Leu Gln Gly Lys Leu Ala Ala Ala Gly Val Glu Thracagtcgtct catgtgccga cgccgaccgt gagacgctcg cccggctcat cgccgagacc 1200Thr Val Val Ser Cys Ala Asp Ala Asp Arg Glu Thr Leu Ala Arg Leu Ile Ala Glu Thrccgcgggaac agccgctgac cgccgtcgtg cacgccgccg acgctccatgg accagtgcc 1260Pro Arg Glu Gln Pro Leu Thr Ala Val Val His Ala Ala Asp Ala Pro Trp Thr Ser Alagtcgccgaca ccggccacgc cgacctcacc gaggtcttcg cgggcaaggtc gacaccgct 1320Val Ala Asp Thr Gly His Ala Asp Leu Thr Glu Val Phe Ala Gly Lys Val Asp Thr Alagtgtggctcg acgaactgtt caccggcacc gacgccgccc cgctcgacgc cttcgtggtc 1380Val Trp Leu Asp Glu Leu Phe Thr Gly Thr Asp Ala Ala Pro Leu Asp Ala Phe Val Valttctcctcga tcgccggcat ctggggcggc ggtggccagg gtgtctccgg cgcggccggc 1440Phe Ser Ser Ile Ala Gly Ile Trp Gly Gly Gly Gly Gln Gly Val Ser Gly Ala Ala Glygcggtcctgg acgcccttgt cgatcggcgc cgcggccggg gactcgcggc cacctcgatc 1500Ala Val Leu Asp Ala Leu Val Asp Arg Arg Arg Gly Arg Gly Leu Ala Ala Thr Ser Ilegcctggggag ccctcgacgg gatcggcctc ggcatggatg aggcggccgc cgcgcagctg 1560Ala Trp Gly Ala Leu Asp Gly Ile Gly Leu Gly Met Asp Glu Ala Ala Ala Ala Gln Leucgccgccgcg gtgtcctgcc gatggcccat caggtcgccg tgaccgcgtt cgaacaggcc 1620Arg Arg Arg Gly Val Leu Pro Met Ala His Gln Val Ala Val Thr Ala Phe Glu Gln Alagcggaggcac gggagaaggc cgtgacggtt gccgacatgg attgggaagc gttcatcccg 1680Ala Glu Ala Arg Glu Lys Ala Val Thr Val Ala Asp Met Asp Trp Glu Ala Phe Ile Progcgttcacct ccgcacgagt cagcccgctc ttcgccgactt gcccgaagcc gcggccgca 1740Ala Phe Thr Ser Ala Arg Val Ser Pro Leu Phe Ala Asp Leu Pro Glu Ala Ala Ala Alactgcgctcct cccaacccga tgccgagaac ggcgacatcac ctcatccctg gtcgactct 1800Leu Arg Ser Ser Gln Pro Asp Ala Glu Asn Gly Asp Ile Thr Ser Ser Leu Val Asp Serctgcgggacg tcccccaggc cgaacagaac cgtctcctgc tccggctggt ctgtgggcag 1860Leu Arg Asp Val Pro Gln Ala Glu Gln Asn Arg Leu Leu Leu Arg Leu Val Cys Gly Glngccgcgaccg tcctcggaca cagcagtggg gagagcatcg gtccgctcca gtccttccag 1920Ala Ala Thr Val Leu Gly His Ser Ser Gly Glu Ser Ile Gly Pro Leu Gln Ser Phe Glngaggtcggct tcgactcgct cggcgccgtc aacctccgca acagcctgca cgtcgccacc 1980Glu Val Gly Phe Asp Ser Leu Gly Ala Val Asn Leu Arg Asn Ser Leu His Val Ala Thrggtctacgac tgcccgcgac actcgtcttc gactacccgac cccggacgc cgtcgtcggc 2040Gly Leu Arg Leu Pro Ala Thr Leu Val Phe Asp Tyr Pro Thr Pro Asp Ala Val Val Glyttcctgcgct ccgagctgct gacggaaacg agcgacgacct ggaagggcg ggaggacgac 2100Phe Leu Arg Ser Glu Leu Leu Thr Glu Thr Ser Asp Asp Leu Glu Gly Arg Glu Asp Aspctgcgacgcg tgctcgcaca ggtcccgctc tcccgccttcg ggaggccgg ccttctcgac 2160Leu Arg Arg Val Leu Ala Gln Val Pro Leu Ser Arg Leu Arg Glu Ala Gly Leu Leu Aspacgctgctca gcctgggcga ctccgtggac ggctccgtccc cgaggcggn ggcgcccgag 2220Thr Leu Leu Ser Leu Gly Asp Ser Val Asp Gly Ser Val Pro Glu Ala XXX Ala Pro Gluccggccccgg cggcgcccgc cgccgaggac gcagcccgtga tcgacgtga tggacgtcgc 2280Pro Ala Pro Ala Ala Pro Ala Ala Glu Asp Ala Ala Arg Asp Arg Arg Asp Gly Arg Argggacctcgta aagcgcgctc tgggcagcaa ccccaactgac t 2322Gly Pro Arg Lys Ala Arg Ser Gly Gln Gln Pro Gln Leu Thr該基因的氨基酸序列同源分析顯示1-39位氨基酸編碼PKS甲基丙二?；D(zhuǎn)移酶；231-687位氨基酸編碼PKS酮基還原酶(KR)、酰基載體蛋白(ACP)及PKS裝配結(jié)構(gòu)域。與利福霉素的I型PKS模塊1-3一致性59％，相似性66％。
9、ShnD GenBank收錄號AF521897，由2973bp DNA序列組成，編碼991個氨基酸序列。atgacggcac cggacgagca gatcgtcgac gcactgcgtg cctcgctcaa ggagaacatg 60Met Thr Ala Pro Asp Glu Gln Ile Val Asp Ala Leu Arg Ala Ser Leu Lys Glu Asn Metcggctccaac aggagaacca gcgtctctcc gagtcctcgg ccgagcccat cgcgatcgtg 120Arg Leu Gln Gln Glu Asn Gln Arg Leu Ser Glu Ser Ser Ala Glu Pro Ile Ala Ile Valtcratggctt gtcggtacgc gggcggcata cgcaaccccg aggacctctg gcgggtggtg 180XXX Met Ala Cys Arg Tyr Ala Gly Gly Ile Arg Asn Pro Glu Asp Leu Trp Arg Val Valaacgacggca ccgacgtcta cacctccttc cccgagaacc gcggctggga cctggagggc 240Asn Asp Gly Thr Asp Val Tyr Thr Ser Phe Pro Glu Asn Arg Gly Trp Asp Leu Glu Glyatctaccacc ccgacccgga caaccccggc acgacgtacg tccgcgaggg tgcgttcctg 300Ile Tyr His Pro Asp Pro Asp Asn Pro Gly Thr Thr Tyr Val Arg Glu Gly Ala Phe Leucacgacgcca acctgttcga cgccgggttg ttcgggatct cgccgcttga ggcgctagcg 360His Asp Ala Asn Leu Phe Asp Ala Gly Leu Phe Gly Ile Ser Pro Leu Glu Ala Leu Alaatggaacctc aacagcggca gcttctcgag atctgctggg aggccctcga acgagccggc 420Met Glu Pro Gln Gln Arg Gln Leu Leu Glu Ile Cys Trp Glu Ala Leu Glu Arg Ala Glyatcgacccgc actccgtacg cggcgccgac atcggcgtat acgccggtct ggtccaccag 480IleAspProHisSerValArgGlyAlaAspIleGlyValTyrAlaGlyLeuValHisGlngactacgcgc ccgacctcag cggcctcgaa ggctacctca gcctggagcg tgctctgggc 540Asp Tyr Ala Pro Asp Leu Ser Gly Leu Glu Gly Tyr Leu Ser Leu Glu Arg Ala Leu Glyagcgcgggcg gcatcgcctc gggacgggtc gcctacacac tcggcctcga aggcccggcc 600Ser Ala Gly Gly Ile Ala Ser Gly Arg Val Ala Tyr Thr Leu Gly Leu Glu Gly Pro Alagtcaccgtcg acaccatgtg ctcctccacc ctggtcgccg tgcacgtggc cacacaggcg 660Val Thr Val Asp Thr Met Cys Ser Ser Thr Leu Val Ala Val His Val Ala Thr Gln Alacttcggcgcg gtgagtgcgc catggccctg gccggcggcg cgaccgtcat gtcgaccccc 720Leu Arg Arg Gly Glu Cys Ala Met Ala Leu Ala Gly Gly Ala Thr Val Met Ser Thr Proggagggttca tcggcttcgc ccggcagcgc gccctcgcct tcgacggccg ctgcaagtcg 780Gly Gly Phe Ile Gly Phe Ala Arg Gln Arg Ala Leu Ala Phe Asp Gly Arg Cys Lys Sertacggggccg ccgcggacgg ctccagctgg gccgagggcg ccggtgtcgt cctcctcgag 840Tyr Gly Ala Ala Ala Asp Gly Ser Ser Trp Ala Glu Gly Ala Gly Val Val Leu Leu Glucggctgtcgg acgcgcgccg caacggacac cgggtcctcg cggtgatccg cggctccgcc 900Arg Leu Ser Asp Ala Arg Arg Asn Gly His Arg Val Leu Ala Val Ile Arg Gly Ser Alactcaaccagg acggcgcctc caacggtctg acggcgccca atggcccggc gcagcggcgc 960Leu Asn Gln Asp Gly Ala Ser Asn Gly Leu Thr Ala Pro Asn Gly Pro Ala Gln Arg Arggtcatccgca aggcgctgga gaacgccggc ctcaccacag ccgacatcga catggtcgag 1020Val Ile Arg Lys Ala Leu Glu Asn Ala Gly Leu Thr Thr Ala Asp Ile Asp Met Val Gluggccacggca ccggcaccgt tctcggcgac ccgatcgagg cccaggccct gatcgccacg 1080Gly His Gly Thr Gly Thr Val Leu Gly Asp Pro Ile Glu Ala Gln Ala Leu Ile Ala Thrtacggccagg accggcccga gggccggccg ctatggctcg gctcggtcaa gtcggtgatc 1140Tyr Gly Gln Asp Arg Pro Glu Gly Arg Pro Leu Trp Leu Gly Ser Val Lys Ser Val Ileggacacaccc agagcggctc cggcgtggcc ggactgatca acgcggtgca ggcgctcagg 1200Gly His Thr Gln Ser Gly Ser Gly Val Ala Gly Leu Ile Asn Ala Val Gln Ala Leu Argcacggcgtca tgcccgccac ccggcacgtc gacgccccca acccgcaggt ggactggtcg 1260His Gly Val Met Pro Ala Thr Arg His Val Asp Ala Pro Asn Pro Gln Val Asp Trp Sergcgggtgcgg tggagctgct gaccgaggcc cgcgcgtggc cggagctggg ccggccacgc 1320Ala Gly Ala Val Glu Leu Leu Thr Glu Ala Arg Ala Trp Pro Glu Leu Gly Arg Pro Argcgggccggtg tgtcctcgtt cggcgccagc ggtacgaacg cccacatgat cctggaacag 1380Arg Ala Gly Val Ser Ser Phe Gly Ala Ser Gly Thr Asn Ala His Met Ile Leu Glu Glngcgccggaag aacccgctgc ggagtccccc tccgccccag cgctcgatgg agtggtaccg 1440Ala Pro Glu Glu Pro Ala Ala Glu Ser Pro Ser Ala Pro Ala Leu Asp Gly Val Val Proctggtgctgt cggcggctac ggccgcctcg ctgaccggcc aggcggagcg actggggtcg 1500Leu Val Leu Ser Ala Ala Thr Ala Ala Ser Leu Thr Gly Gln Ala Glu Arg Leu Gly Serttcctcgagg cgtccggcac ggtcgcgctc gccgatgtgg cggccgcact ggtcaccggc 1560Phe Leu Glu Ala Ser Gly Thr Val Ala Leu Ala Asp Val Ala Ala Ala Leu Val Thr Glycgggcgtcgc tggcccagcg cgcggtcgtc gtgaccgact cgcccgagga ggccctggca 1620Arg Ala Ser Leu Ala Gln Arg Ala Val Val Val Thr Asp Ser Pro Glu Glu Ala Leu Alaggtctcggtg cgctggctcg tggtgaggat gttcgtgggg tggttgctgg tggtggggtg 1680Gly Leu Gly Ala Leu Ala Arg Gly Glu Asp Val Arg Gly Val Val Ala Gly Gly Gly Valaggtcgggtg gggacggcaa ggttgtgttg gtgtttccgg gtcagggttc gccgtgggtt 1740Arg Ser Gly Gly Asp Gly Lys Val Val Leu Val Phe Pro Gly Gln Gly Ser Pro Trp Valggtatggggc gtgagttgtt ggagtgttcg gaggtgtttg cggcgcgggt gggggagtgt 1800Gly Met Gly Arg Glu Leu Leu Glu Cys Ser Glu Val Phe Ala Ala Arg Val Gly Glu Cysgcggtggcgt tggagcggtg ggtggattgg tcgttggtgg atgtgttgcg gggggattgt 1860Ala Val Ala Leu Glu Arg Trp Val Asp Trp Ser Leu Val Asp Val Leu Arg Gly Asp Cysccggttgagt tttttgagcg tgaggatgtg cggcagccgg cgagttttgc ggtgatggtg 1920Pro Val Glu Phe Phe Glu Arg Glu Asp Val Arg Gln Pro Ala Ser Phe Ala Val Met Valggtttggccg cggtgtggga gtcggtgggt gtggtggcgg atgcggtggt gggtcattcg 1980Gly Leu Ala Ala Val Trp Glu Ser Val Gly Val Val Ala Asp Ala Val Val Gly His Serggtggtgagg ttgctgctgc gtgtgtgtcg gktgcgttgt cgttggagga cgctgttcgg 2040Gly Gly Glu Val Ala Ala Ala Cys Val Ser XXX Ala Leu Ser Leu Glu Asp Ala Val Arggttgtggcgg tccggagcaa gaccatttcc ggtgttctct cgggtcgggg tggtatggcg 2100Val Val Ala Val Arg Ser Lys Thr Ile Ser Gly Val Leu Ser Gly Arg Gly Gly Met Alatcggtggggt tgtcggagga ggaggcggtt gctcggctac agcagtggga tggtcgggtc 2160Ser Val Gly Leu Ser Glu Glu Glu Ala Val Ala Arg Leu Gln Gln Trp Asp Gly Arg Valgagatcgggg cggtcaacag tccgtcctcg gtggccatca ccgctgacac cgaagccctc 2220Glu Ile Gly Ala Val Asn Ser Pro Ser Ser Val Ala Ile Thr Ala Asp Thr Glu Ala Leugacgaagcca tcgagacttt ggaggaccag ggcgtccgcg tacgccgcat cgcgatcgac 2280Asp Glu Ala Ile Glu Thr Leu Glu Asp Gln Gly Val Arg Val Arg Arg Ile Ala Ile Asptacgcctcgc actcccggca cgtcggggct gtccaggaga tcctgaacga ggcattcgcc 2340Tyr Ala Ser His Ser Arg His Val Gly Ala Val Gln Glu Ile Leu Asn Glu Ala Phe Alagacatccgca gccaagctcc taccgtgcca ttcctctcca ccgccaccgg cgagtggatc 2400Asp Ile Arg Ser Gln Ala Pro Thr Val Pro Phe Leu Ser Thr Ala Thr Gly Glu Trp Ilecgcgaggcgg gtgccctgga cggcagctac tggtaccgca acctgcgcag ccaggtccgc 2460Arg Glu Ala Gly Ala Leu Asp Gly Ser Tyr Trp Tyr Arg Asn Leu Arg Ser Gln Val Argttcggccccg cgatcgccga cctgctggcc gacggccaca ccgtgttcgt ggagtccagc 2520Phe Gly Pro Ala Ile Ala Asp Leu Leu Ala Asp Gly His Thr Val Phe Val Glu Ser Sergcccaccccg tcctggtcca gccgatcagc gaggtcgtgg ccggcgccga ggcagaggcc 2580Ala His Pro Val Leu Val Gln Pro Ile Ser Glu Val Val Ala Gly Ala Glu Ala Glu Alagtcgtgaccg gctccctgcg ccgtcacgag ggaggtccgc gccgcctgtt cacttcgatg 2640Val Val Thr Gly Ser Leu Arg Arg His Glu Gly Gly Pro Arg Arg Leu Phe Thr Ser Metgccgacctct tcgtccgagg cacccacgtc gactggagcg gcgtcctcgc ggccggagcc 2700Ala Asp Leu Phe Val Arg Gly Thr His Val Asp Trp Ser Gly Val Leu Ala Ala Gly Alagatgcccgcc gcgtcgacct tccgacgtac gcctttgatc acaagaacta ctggatggag 2760Asp Ala Arg Arg Val Asp Leu Pro Thr Tyr Ala Phe Asp His Lys Asn Tyr Trp Met Gluctggccggta ccgccaacga tgtcgcctcg ctcggcttgt cgggggctga tcatccgttg 2820Leu Ala Gly Thr Ala Asn Asp Val Ala Ser Leu Gly Leu Ser Gly Ala Asp His Pro Leuctgggtgcgg tggttccggt gccggagacg agcggagtgt tgtgtacgtc gcggttgtcg 2880Leu Gly Ala Val Val Pro Val Pro Glu Thr Ser Gly Val Leu Cys Thr Ser Arg Leu Sercttcgtacac atccgtggct tgcggatcac gctgtgggcc ggtgtcgtgc ttgtccctgg 2940Leu Arg Thr His Pro Trp Leu Ala Asp His Ala Val Gly Arg Cys Arg Ala Cys Pro Trpcactgctttg gtggagttgg tggtgcgtgc ggg 2973His Cys Phe Gly Gly Val Gly Gly Ala Cys Gly34-285位的氨基酸編碼PKS的KS-N端結(jié)構(gòu)域；293-460位的氨基酸編碼PKS的KS-C端結(jié)構(gòu)域，34-460位氨基酸編碼PKS的β酮酰基合酶(KS)；571-873位氨基酸編碼酰基轉(zhuǎn)移酶(AT)；199-238位氨基酸編碼硫酯酶(TE)的N端結(jié)構(gòu)域。與利福霉素的I型PKS一致性66％，相似性76％。 該基因的氨基酸序列分析顯示含有4個保守的結(jié)構(gòu)域(圖8)9、ShnE GenBank收錄號AF521897，不完整的ORF，現(xiàn)獲得1404bp序列，編碼468個氨基酸序列。atgcccgccg cgtcgacctt ccgacgtacg cctttgatca caagaactac tggatggagc 60Met Pro Ala Ala Ser Thr Phe Arg Arg Thr Pro Leu Ile Thr Arg Thr Thr Gly Trp Sertggccggtac cgccaacgat gtcgcctcgc tcggcttgtc gggggctgat catccgttgc 120Trp Pro Val Pro Pro Thr Met Ser Pro Arg Ser Ala Cys Arg Gly Leu Ile Ile Arg Cystgggtgcggt ggttccggtg ccggagacga gcggagtgtt gtgtacgtcg cggttgtcgc 180Trp Val Arg Trp Phe Arg cys Arg Arg Arg Ala Glu cys cys Val Arg Arg Gly cys Argttcgtacaca tccgtggctt gcggatcacg ctgtgggccg gtgtcgtgct tgtccctggc 240Phe Val His Ile Arg Gly Leu Arg Ile Thr Leu Trp Ala Gly Val Val Leu Val Pro Glyactgctttgg tggagttggt ggtgcgtgcg ggtgacaagg tgggctgcgg cacgttggag 300Thr Ala Leu Val Glu Leu Val Val Arg Ala Gly Asp Lys Val Gly Cys Gly Thr Leu Glugaattggtca tcgagacgcc gcttgtcgta cccgcgcaag ggagtatgcg cgttcagttc 360Glu Leu Val Ile Glu Thr Pro Leu Val Val Pro Ala Gln Gly Ser Met Arg Val Gln Phegcggtgggcg gccctgagga gaacggcgcg cgttcggtgg ccgtgtactc ggctcgtgat 420Ala Val Gly Gly Pro Glu Glu Asn Gly Ala Arg Ser Val Ala Val Tyr Ser Ala Arg Aspgacgacggtc gcggcaccgg tatcgatggt tggacccgtc acgccgccgg cactctgacg 480Asp Asp Gly Arg Gly Thr Gly Ile Asp Gly Trp Thr Arg His Ala Ala Gly Thr Leu Thrgcggctgctg tccctgctga tggtttcgat ttcacggtgt ggccgccggt cggtgcggag 540Ala Ala Ala Val Pro Ala Asp Gly Phe Asp Phe Thr Val Trp Pro Pro Val Gly Ala Glucgggtgtcgt tcgatgcggt cgggttctat gaggagatgg cgggccgcgg ctatgtgtac 600Arg Val Ser Phe Asp Ala Val Gly Phe Tyr Glu Glu Met Ala Giy Arg Gly Tyr Val Tyrggtccggcgt tccagggttt gcgtggggtg tggcggcggg gcgaagaggt gttcgccgag 660Gly Pro Ala Phe Gln Gly Leu Arg Gly Val Trp Arg Arg Gly Glu Glu Val Phe Ala Glugtcgctctgc cggacgagca gcatggtgag gcgagccgct tcgggttgca cccggcgttg 720Val Ala Leu Pro Asp Glu Gln His Gly Glu Ala Ser Arg Phe Gly Leu His Pro Ala Leuctcgacgccg ccttgcagag cgggctcgtc cggccggccg atgccggggt ggatatgcgt 780Leu Asp Ala Ala Leu Gln Ser Gly Leu Val Arg Pro Ala Asp Ala Gly Val Asp Met Arggtgccgttcg cctggaacgg gctgcgcctg catgccgcgg gtgcctcgga gttgcgggtg 840Val Pro Phe Ala Trp Asn Gly Leu Arg Leu His Ala Ala Gly Ala Ser Glu Leu Arg Valcggacggtgc cgtccgggcc ggacgcggtg tcgttgcagg cggccgacgg ggccggcggt 900Arg Thr Val Pro Ser Gly Pro Asp Ala Val Ser Leu Gln Ala Ala Asp Gly Ala Gly Glyccggtgctga gcctggagtc gctggttgcc cgggcggtgg acgtggagca actggatcgg 960Pro Val Leu Ser Leu Glu Ser Leu Val Ala Arg Ala Val Asp Val Glu Gln Leu Asp Argatggcgactg atgacggtcg cgacgcgctg ttcgaggtgg actggagcga actgcccgcg 1020Met Ala Thr Asp Asp Gly Arg Asp Ala Leu Phe Glu Val Asp Trp Ser Glu Leu Pro Alacctgcttcga gcgtggagtc tctgccgccg tcggcgctgg tggcttcggc cgaggacgtg 1080Pro Ala Ser Ser Val Glu Ser Leu Pro Pro Ser Ala Leu Val Ala Ser Ala Glu Asp Valacggatctgg cagatgccgc ggtggttcct gcggtagcgg ttcttgaggc tgttggcggt 1140Thr Asp Leu Ala Asp Ala Ala Val Val Pro Ala Val Ala Val Leu Glu Ala Val Gly Glygacggcgagc acgacgcgct tgccctgacc gtcagggtgc tggaggtcgt ccaggcgtgg 1200Asp Gly Glu His Asp Ala Leu Ala Leu Thr Val Arg Val Leu Glu Val Val Gln Ala Trpttcgctgctg cgggtctggc ggagtcccgg ctggtggtgg tcacacgggg tgcggttccg 1260Phe Ala Ala Ala Gly Leu Ala Glu Ser Arg Leu Val Val Val Thr Arg Gly Ala Val Progtcggcggtg agggaaatgt cgccgatccc gctggtgctg cggtgtgggg tctggttcgg 1320Val Gly Gly Glu Gly Asn Val Ala Asp Pro Ala Gly Ala Ala Val Trp Gly Leu Val Arggcggcccagg ccgagaaccc ggaccggatc gtcctgctcg atctcgctgc cgacgtcgat 1380Ala Ala Gln Ala Glu Asn Pro Asp Arg Ile Val Leu Leu Asp Leu Ala Ala Asp Val Aspatgggatcgg ttctgcctgc cgta1404Met Gly Ser Val Leu Pro Ala Val Leu與利福霉素的I型PKS模塊4-6一致性51％，相似性62％，該基因的氨基酸序列分析顯示74-465位編碼氨基酸編碼脫水酶(DH)。
pCGBA3組基因簇序列分析表明，其AHBA編碼基因與已知的萘醌類安莎霉素利福霉素、napthomycin的同源性高于苯醌類安莎霉素，而且從基因轉(zhuǎn)錄方向及結(jié)構(gòu)上更接近萘醌型安莎類抗生素，表明本發(fā)明在S.hygroscopicus17997中發(fā)現(xiàn)了編碼格爾德霉素以外的一個萘醌類安莎霉素生物合成基因簇存在。四.S.hygroscopicus 17997中萘醌型安莎霉素生物學活性的測定本發(fā)明利用基因阻斷技術(shù)破壞S.hygroscopicus 17997中負責格爾德霉素生物合成基因(圖9)，以檢測本發(fā)明所發(fā)現(xiàn)的該菌產(chǎn)生格爾德霉素以外的另一個萘醌類安莎霉素的生物學活性。
將S.hygroscopicus 17997中與格爾德霉素生物合成相關(guān)基因序列，插入質(zhì)粒載體如噬菌體載體KC515[Microbiol.Rev.1980，Jun44(2)206-229]，構(gòu)建重組質(zhì)粒phi203(圖10)，轉(zhuǎn)染S.hygroscopicus 17997孢子，獲得溶源菌，分離提取溶源菌總DNA，用限制性酶如BamHI酶切后，以KC515載體上硫鏈絲菌素抗性標記基因為探針，進行Southern雜交分析，證明溶源菌中所含與格爾德霉素生物合成相關(guān)基因序列已和載體一起整合至染色體，致使格爾德霉素生物合成基因受到破壞。溶源菌經(jīng)發(fā)酵培養(yǎng)28℃，7天，用二氯甲烷抽提發(fā)酵上清液，提取液濃縮至干，以80％甲醇溶解，進行HPLC檢測，證明該溶源菌確實已不再產(chǎn)生格爾德霉素。以單純皰疹病毒I型感染的VERO細胞為模型，測定溶源菌發(fā)酵液對病毒的活性，結(jié)果見下表。
溶源菌抗病毒活性

由表中結(jié)果可見不產(chǎn)生格爾德霉素的溶源菌，所產(chǎn)生的另一萘醌型安莎類化合物具有明顯的抗皰疹病毒的活性。
以下實施例對本發(fā)明而言，僅為說明性的，而非限制性的。
實施例一從生物體篩選安莎類化合物利用生物學軟件Vector NTIsuit6.0程序，對NCBI數(shù)據(jù)庫中所有登錄的含AHBA結(jié)構(gòu)的抗生素利福霉素B，ansatrieninA，napthomycin，絲裂霉素C，ansamitocin等的AHBA合酶編碼序列分析后，根據(jù)其氨基酸序列的保守區(qū)設(shè)計一對簡并引物，如本發(fā)明中所述的上游引物及下游引物，以產(chǎn)生安莎類抗生素格爾德霉素產(chǎn)生菌S.hygroscopicus17997基因組DNA為模板，在LATaq酶作用下進行PCR反應，擴增出755bp特異性條帶。對PCR產(chǎn)物測序后與NCBI數(shù)據(jù)庫基因比較，證實其與安莎類化合物AHBA基因同源，以此為探針與其他生物體菌落或機體進行雜交，對顯示陽性的生物體產(chǎn)物，按安莎類化合物性質(zhì)分離、鑒別即可得到安莎類化合物。
實施例二安莎類化合物生物合成基因的獲取利用生物學軟件Vector NTIsuit6.0程序，對NCBI數(shù)據(jù)庫中所有登錄的含AHBA結(jié)構(gòu)的抗生素利福霉素B，ansatrieninA，napthomycin，絲裂霉素C，ansamitocin等的AHBA合酶編碼序列分析后，根據(jù)其氨基酸序列的保守區(qū)設(shè)計一對簡并引物，如本發(fā)明中所述的上游引物及下游引物，以產(chǎn)生安莎類抗生素如格爾德霉素產(chǎn)生菌S.hygroscopicus17997基因組DNA為模板，在LATaq酶作用下進行PCR反應，擴增出特異性條帶對PCR產(chǎn)物測序后與NCBI數(shù)據(jù)庫基因比較，證實其與安莎類化合物AHBA基因同源，以此為探針與其他生物體菌落或機體進行雜交，提取分離陽性菌落或機體的DNA，用柯斯質(zhì)粒載體建立基因組文庫，用AHBA基因探針與基因文庫進行雜交，對陽性柯斯質(zhì)粒外源片段進行測序，測序結(jié)果與NCBI數(shù)據(jù)庫進行同源性比較，即可獲得安莎類化合物生物合成基因。發(fā)明效果本發(fā)明通過負責AHBA這一安莎類抗生素特異結(jié)構(gòu)的AHBA合酶基因保守序列，從格爾德霉素產(chǎn)生菌發(fā)現(xiàn)了一個新的安莎類化合物生物合成基因簇，并證明該新化合物具有抗病毒活性，說明利用分子生物學手段，克隆某一類抗生素特異結(jié)構(gòu)的生物合成基因保守序列，進而篩選獲得這一類化合物，是一個可行且特異的新的篩選方法。這一思路有可能用于其他類型化合物的篩選，如利用異青霉素合成酶(IPNS)基因保守序列，可以篩選獲得青霉素、頭孢菌素等β-內(nèi)酰胺類化合物；利用6-脫氧己糖酶(6-DOH)基因保守序列，可以篩選獲得鏈霉素、卡那霉素等氨基糖苷類化合物等。


圖1 以AHBA保守序列設(shè)計引物所獲PCR產(chǎn)物電泳其中1 分子量標記2 PCR產(chǎn)物755bp3 分子量標記λDNA/HindIII圖2 755bp探針獲得的陽性克隆BamHI酶切凝膠電泳(A)及Southern雜交(B)1.λ/HindIII 2.pCGBA1/BamHI 3.pCGBA2/BamHI4.pCGBA3/BamHI 5.pCGBA4/BamHI 6.pCGBA5/BamHI7.pCGBA6/BamHI 8.pCGBA7/BamHI 9.pCGBA8/BamHI10.pCGBA9/BamHI11.pCGBA10/BamHI 12.pCGBA11/BamHI13.pCGBA12/BamHI圖3 pCGBA3柯斯質(zhì)粒中基因開放閱讀框架示意圖其中shnAI型聚酮合酶shnBI型聚酮合酶shnCI型聚酮合酶shnDI型聚酮合酶shnEI型聚酮合酶shnN酰胺合成酶shnQ氨基脫氫奎尼酸合成酶shnSAHBA合酶shnO氧化還原酶shnP磷酸化酶圖4 ShnA 聚酮合酶基因結(jié)構(gòu)域其中adh-short 短鏈脫氫酶pp-binding 磷酸泛酰巰基乙胺的附著位點圖5 shnS AHBA 合酶基因結(jié)構(gòu)域其中DegT-DnrJ-EryC 信號轉(zhuǎn)導系統(tǒng)中的傳感蛋白Cys-Met-Meta-PP 半胱氨酸/甲硫氨酸磷酸吡哆醛(PLP)依賴酶Aminotran-1-2 氨基轉(zhuǎn)移酶家族圖6 ShnB 聚酮合酶基因結(jié)構(gòu)域其中 ketoacyl-syntβ酮?；厦窷端結(jié)構(gòu)域硫酯酶N端結(jié)構(gòu)域β酮?；厦窩端結(jié)構(gòu)域acyl-transf ?；D(zhuǎn)移酶結(jié)構(gòu)域adh short短鏈脫氫酶3 Beta HSD β羥基固醇脫氫酶/異構(gòu)酶家族 磷酸泛酰巰基乙胺的附著位點圖7 ShnO 氧化還原酶基因結(jié)構(gòu)域其中 acetylald-DH 乙醛脫氫酶家族PDH 預苯酸脫氫酶家族Polysacc-synt-2 多聚糖生物合成蛋白圖8 ShnD 基因結(jié)構(gòu)域其中ketoacyl-syntPKS的KS-N端結(jié)構(gòu)域；ketoacyl-synt-c PKS的KS-C端結(jié)構(gòu)域acyl-transf ?；D(zhuǎn)移酶硫酯酶的N端結(jié)構(gòu)域。
圖9 基因阻斷技術(shù)破壞目的基因示意圖其中targetABC目的基因＿tsr＿ 載體上硫鏈絲菌素抗性基因代表載體部分圖10 格爾德霉素生物合成基因與噬菌體載體KC515重組質(zhì)粒的構(gòu)建序列表<110>中國醫(yī)學科學院醫(yī)藥生物技術(shù)研究所<120>利用3-氨基-5-羥基苯甲酸(AHBA)合酶基因保守序列篩選安莎類化合物的方法<160>21<170>PatentIn version 3.1<210>1<211>755<212>DNA<213>吸水鏈霉菌(Streptomyces hygroscopicus)<220><221>misc_feature<222>(1)..(755)<223>partial CDS<400>1ttcgagcagg agttcgcgga cttccacggc gccccacacg ctttggccgt caccaacggc 60acccacgccc tggagttggc gttgcagtgt ctgggcgtcg ggccgggcac cgaggtcatc120gtgccggcct tcaccttcat ctcctcctcc caggccgctc agcggctggg agcggttgcc180gtccccgtcg acgtcgatct cgatacctac aacatcgacg tggctgccgc ggcttccgcc240gtcacccccc tcaccaaggc gatcatgcct gtgcacatgg cggggctcat cgccgacatg300gacgcgctcg gcgaactctc cgccgacacc ggtgtgcctc ttctccagga cgccgcccac360gcacacggtg cccgctggca gggcaaacgg gtgggcgagt tgggtacggt cgcctcgttc420agcttccaga acggcaagct gatgaccgcc ggcgagggcg gtgcgctgct cctgcccgac480gaggagacct acgaggccgc gttcctgcgg cacagttgtg gccggtcacg taccgaccgc540cgatacatgc accagaccgc cggcacgaac atgcggctca acgagttctc cgcggccgtg600ctccgcgccc agctgggccg cctcgacgcc cagatcacgc tccgcgatca gcgctggacg660ctgctgtccc ggctgctcgg tgagatcgac agcgtcgtac cccagggcag cgacccgcgc720gccgaccgga actcccacta catggccatg gtccg 755<210>2<211>1455<212>DNA<213>吸水鏈霉菌(Streptomyces hygroscopicus)<220><221>CDS<222>(1)..(1455)<223>shnA<220><221>misc_feature<222>(792，815，816，817)<223>k＝g或t；m＝a或c；s＝g或c<400>2gtg ggc gta ctg gcg gat gtg cac gca tcc ggc gag ccg cag gtc gca 48Val Gly Val Leu Ala Asp Val His Ala Ser Gly Glu Pro Gln Val Ala1 5 10 15ctc cga tcg ggc gcg gtc ctc gtc ccg cgt ctc gct cgt gtc gcc gat 96Leu Arg Ser Gly Ala Val Leu Val Pro Arg Leu Ala Arg Val Ala Asp20 25 30acg gac cgg gct tcc acc ggc cgt cgt ctc gac ccc gat ggc acc gcg144Thr Asp Arg Ala Ser Thr Gly Arg Arg Leu Asp Pro Asp Gly Thr Ala35 40 45ctg ata acc ggt ggt acc ggt gcg ctc ggt gcg ctg gtg gcc cgg cat192Leu Ile Thr Gly Gly Thr Gly Ala Leu Gly Ala Leu Val Ala Arg His50 55 60ctg gtc gtc gag cac aag atc cgg agt ctg gtc ctg gta agc cgt cgg240Leu Val Val Glu His Lys Ile Arg Ser Leu Val Leu Val Ser Arg Arg65 70 75 80ggc ccg gac gcc ccg ggg gcc gcc gat ctc gac gcg gag ctg act gcc288Gly Pro Asp Ala Pro Gly Ala Ala Asp Leu Asp Ala Glu Leu Thr Ala85 90 95ctg ggt gcc cgc gtg cga att gtc gcc tgc gac atc gcg gac cgc gag336Leu Gly Ala Arg Val Arg Ile Val Ala Cys Asp Ile Ala Asp Arg Glu100 105 110gcg gcc ggg gaa ctg atc gcc tct gta ccg cgg gac gcg ccg ctc act384Ala Ala Gly Glu Leu Ile Ala Ser Val Pro Arg Asp Ala Pro Leu Thr115 120 125gcc gtg gtg cac acg gcc ggt gtg ctg gac ggc ggc gtg gtc act gcc432Ala Val Val His Thr Ala Gly Val Leu Asp Gly Gly Val Val Thr Ala130 135 140ctt acg ccg gag cgg ctc gat gcc gtt ctc cgg ccg aag gcg gac gcc480Leu Thr Pro Glu Arg Leu Asp Ala Val Leu Arg Pro Lys Ala Asp Ala145 150 155 160gct ctg gtc ctg gac gag ctg acc cgc cac ctg gac gtg gcg gcc ttc528Ala Leu Val Leu Asp Glu Leu Thr Arg His Leu Asp Val Ala Ala Phe165 170 175gtc ctg ttc tcc tcc gcc gcc gga acg ttc ggc aac ccc ggc cag gga576Val Leu Phe Ser Ser Ala Ala Gly Thr Phe Gly Asn Pro Gly Gln Gly180 185 190aac ctg gcc gcc tcg aac gcg tat ctt gac gcg ctg gcg gta cga cgc624Asn Leu Ala Ala Ser Asn Ala Tyr Leu Asp Ala Leu Ala Val Arg Arg195 200 205cgg act gcc ggg ctg ccc gcc aca tct gtt gcc tgg ggg gtg tgg gac672Arg Thr Ala Gly Leu Pro Ala Thr Ser Val Ala Trp Gly Val Trp Asp210 215 220cag acc ggc atc agc ggg gac ctt ggc gtg gcc gat cag cgg agg atg720Gln Thr Gly Ile Ser Gly Asp Leu Gly Val Ala Asp Gln Arg Arg Met225 230 235 240gcc cgg tgg ggc ctg gcg gca cac tcc gcc cag gag ggt ctg gag ctg768Ala Arg Trp Gly Leu Ala Ala His Ser Ala Gln Glu Gly Leu Glu Leu245 250 255ttc gac gcg gcg ttg cgg gca gak gac gcg gtg ctc gtg gcc gcg ams816Phe Asp Ala Ala Leu Arg Ala Xaa Asp Ala Val Leu Val Ala Ala Xaa260 265 270mtg aac ttc gcc ggg ctg cgc gcc cag gcc gcc tcc gaa ccc gta cac864Xaa Asn Phe Ala Gly Leu Arg Ala Gln Ala Ala Ser Glu Pro Val His275 280 285gta ctg ctg cgg ggc ctc gtg cgg gcc ggc cgt cgc gcc gct cag cag912Val Leu Leu Arg Gly Leu Val Arg Ala Gly Arg Arg Ala Ala Gln Gln290 295 300gca tcc tcc cgc gag ggc ggc ctc gcc gga cag ctg gct gcc acg ccc 960Ala Ser Ser Arg Glu Gly Gly Leu Ala Gly Gln Leu Ala Ala Thr Pro305 310 315 320ccg gtt cag cgg gag cag atc ctg ctg gat ctg gtg cgg cgc gag gtc1008Pro Val Gln Arg Glu Gln Ile Leu Leu Asp Leu Val Arg Arg Glu Val325 330 335gct gcg gtc ctc ggc tat tcg aca cca cgc aag gtc gac ccc gac cgg1056Ala Ala Val Leu Gly Tyr Ser Thr Pro Arg Lys Val Asp Pro Asp Arg340 345 350gcc ttc cag gac gtc ggg ttc acc tcg gtg ctg gcc gtc gaa ctc cgt1104Ala Phe Gln Asp Val Gly Phe Thr Ser Val Leu Ala Val Glu Leu Arg355 360 365aac cga ctc gcc ggg ctc gcg ggg atc cgg ctc ccg gca tcg atc gcc1152Asn Arg Leu Ala Gly Leu Ala Gly Ile Arg Leu Pro Ala Ser Ile Ala370 375 380ttc gac cat ccc aca ccg cgg cgc atg atg cgc cat ctg ctc gcg gaa1200Phe Asp His Pro Thr Pro Arg Arg Met Met Arg His Leu Leu Ala Glu385 390 395 400ctg tgc ccc gag gat ggc agc gag ccg gcg gac cgg gag gat gag atc1248Leu Cys Pro Glu Asp Gly Ser Glu Pro Ala Asp Arg Glu Asp Glu Ile405 410 415cgc agg gcc ctg gcg acc aca cct ctg tcc cgg ttc cgc gag ctt ggg1296Arg Arg Ala Leu Ala Thr Thr Pro Leu Ser Arg Phe Arg Glu Leu Gly420 425 430ctc atg gag cag atc ctg cac ctg gtg gcg cat ccc agt ggt gaa agt1344Leu Met Glu Gln Ile Leu His Leu Val Ala His Pro Ser Gly Glu Ser435 440 445gcc gca gca ccg gac acc gcg gaa ccg aaa cag gac gcc ggg ccg ctg1392Ala Ala Ala Pro Asp Thr Ala Glu Pro Lys Gln Asp Ala Gly Pro Leu450 455 460atc gcg gag atg gac atc gac aac ctt gtg aag cgg gcg atg gaa aag1440Ile Ala Glu Met Asp Ile Asp Asn Leu Val Lys Arg Ala Met Glu Lys465 470 475 480gcc cgg aag ccg tag1455Ala Arg Lys Pro<210>3<211>484<212>PRT<213>吸水鏈霉菌(Streptomyces hygroscopicus)<220><221>misc_feature<222>(264)<223>Xaa＝Glu或Asp.<220><221>misc_feature<222>(272)<223>Xaa＝Lys或Asn或Thr.<220><221>misc_feature<222>(273)<223>Xaa＝Met或Leu.<400>3Val Gly Val Leu Ala Asp Val His Ala Ser Gly Glu Pro Gln Val Ala1 5 10 15Leu Arg Ser Gly Ala Val Leu Val Pro Arg Leu Ala Arg Val Ala Asp20 25 30Thr Asp Arg Ala Ser Thr Gly Arg Arg Leu Asp Pro Asp Gly Thr Ala35 40 45Leu Ile Thr Gly Gly Thr Gly Ala Leu Gly Ala Leu Val Ala Arg His50 55 60Leu Val Val Glu His Lys Ile Arg Ser Leu Val Leu Val Ser Arg Arg65 70 75 80Gly Pro Asp Ala Pro Gly Ala Ala Asp Leu Asp Ala Glu Leu Thr Ala85 90 95Leu Gly Ala Arg Val Arg Ile Val Ala Cys Asp Ile Ala Asp Arg Glu100 105 110Ala Ala Gly Glu Leu Ile Ala Ser Val Pro Arg Asp Ala Pro Leu Thr115 120 125Ala Val Val His Thr Ala Gly Val Leu Asp Gly Gly Val Val Thr Ala130 135 140Leu Thr Pro Glu Arg Leu Asp Ala Val Leu Arg Pro Lys Ala Asp Ala145 150 155 160Ala Leu Val Leu Asp Glu Leu Thr Arg His Leu Asp Val Ala Ala Phe165 170 175Val Leu Phe Ser Ser Ala Ala Gly Thr Phe Gly Asn Pro Gly Gln Gly180 185 190Asn Leu Ala Ala Ser Asn Ala Tyr Leu Asp Ala Leu Ala Val Arg Arg195 200 205Arg Thr Ala Gly Leu Pro Ala Thr Ser Val Ala Trp Gly Val Trp Asp210 215 220Gln Thr Gly Ile Ser Gly Asp Leu Gly Val Ala Asp Gln Arg Arg Met225 230 235 240Ala Arg Trp Gly Leu Ala Ala His Ser Ala Gln Glu Gly Leu Glu Leu245 250 255Phe Asp Ala Ala Leu Arg Ala Xaa Asp Ala Val Leu Val Ala Ala Xaa260 265 270Xaa Asn Phe Ala Gly Leu Arg Ala Gln Ala Ala Ser Glu Pro Val His275 280 285Val Leu Leu Arg Gly Leu Val Arg Ala Gly Arg Arg Ala Ala Gln Gln290 295 300Ala Ser Ser Arg Glu Gly Gly Leu Ala Gly Gln Leu Ala Ala Thr Pro305 310 315 320Pro Val Gln Arg Glu Gln Ile Leu Leu Asp Leu Val Arg Arg Glu Val325 330 335Ala Ala Val Leu Gly Tyr Ser Thr Pro Arg Lys Val Asp Pro Asp Arg340 345 350Ala Phe Gln Asp Val Gly Phe Thr Ser Val Leu Ala Val Glu Leu Arg355 360 365Asn Arg Leu Ala Gly Leu Ala Gly Ile Arg Leu Pro Ala Ser Ile Ala370 375 380Phe Asp His Pro Thr Pro Arg Arg Met Met Arg His Leu Leu Ala Glu385 390 395 400Leu Cys Pro Glu Asp Gly Ser Glu Pro Ala Asp Arg Glu Asp Glu Ile405 410 415Arg Arg Ala Leu Ala Thr Thr Pro Leu Ser Arg Phe Arg Glu Leu Gly420 425 430Leu Met Glu Gln Ile Leu His Leu Val Ala His Pro Ser Gly Glu Ser435 440 445Ala Ala Ala Pro Asp Thr Ala Glu Pro Lys Gln Asp Ala Gly Pro Leu450 455 460Ile Ala Glu Met Asp Ile Asp Asn Leu Val Lys Arg Ala Met Glu Lys465 470 475 480Ala Arg Lys Pro<210>4<211>5319<212>DNA<213>吸水鏈霉菌(Streptomyces hygroscopicus)<220><221>CDS<222>(1)..(5319)<223>shnB<220><221>misc_feature<222>(5017，5023，5027，5043，5044，5047)<223>r＝a或g；y＝c或t；k＝g或t<400>4gtg gac gca cgc cag aac acg gag ata gca gtg gcc agt tca gag agc48Val Asp Ala Arg Gln Asn Thr Glu Ile Ala Val Ala Ser Ser Glu Ser1 5 10 15aag gtc gtc gaa gca ctg cgc gct tca ctg atg gag aac gaa cgg ctg96Lys Val Val Glu Ala Leu Arg Ala Ser Leu Met Glu Asn Glu Arg Leu20 25 30gag agc gag gtc cag agc atc cgc gac agc ctc acc gag ccg atc gcc 144Glu Ser Glu Val Gln Ser Ile Arg Asp Ser Leu Thr Glu Pro Ile Ala35 40 45atc gtc ggc atg gcg tgc cgg ttc ccc ggc ggg gtg tcg tcg ccg gaa 192Ile Val Gly Met Ala Cys Arg Phe Pro Gly Gly Val Ser Ser Pro Glu50 55 60gag ttg tgg gaa ttg atc gcg gac ggc cgt tcc gcg gtc gag gcg ttc 240Glu Leu Trp Glu Leu Ile Ala Asp Gly Arg Ser Ala Val Glu Ala Phe65 70 75 80ccc acc aac cgg ggc tgg gac ctg gag aac ctg tac gac ccg gac ctc288Pro Thr Asn Arg Gly Trp Asp Leu Glu Asn Leu Tyr Asp Pro Asp Leu85 90 95gac cgg ccc ggc acg acg tac gta cgg gag ggc gcg ttc ctg cac gac336Asp Arg Pro Gly Thr Thr Tyr Val Arg Glu Gly Ala Phe Leu His Asp100 105 110gcg ggc gag ttc gac gcc ggc ttc ttc ggc atc tcc caa agc gag acg384Ala Gly Glu Phe Asp Ala Gly Phe Phe Gly Ile Ser Gln Ser Glu Thr115 120 125atg gtc atg gac cca cag cag cgc ctg atg ctg gag aca tct tgg gag432Met Val Met Asp Pro Gln Gln Arg Leu Met Leu Glu Thr Ser Trp Glu130 135 140gcg ttc gaa cgg gcg ggc atc gac ccg gcc gct atg cgt ggc aag aac480Ala Phe Glu Arg Ala Gly Ile Asp Pro Ala Ala Met Arg Gly Lys Asn145 150 155 160gtc ggc gtg ttc gcc ggc atg gcc gcc ggg cag gag tac ggg acc gct528Val Gly Val Phe Ala Gly Met Ala Ala Gly Gln Glu Tyr Gly Thr Ala165 170 175ttc cac agc atc ccc gac gag ctc gag ggc tat gtg atg acc ggc ggt576Phe His Ser Ile Pro Asp Glu Leu Glu Gly Tyr Val Met Thr Gly Gly180 185 190ctg gcg agc gtc ctt tcg gga cgg gtc tcc tat acg ttc gga ttc gag624Leu Ala Ser Val Leu Ser Gly Arg Val Ser Tyr Thr Phe Gly Phe Glu195 200 205ggg ccg gca gtc acg atc gac acc gcc tgc tcc tcg tcc ctg gtg gcc672Gly Pro Ala Val Thr Ile Asp Thr Ala Cys Ser Ser Ser Leu Val Ala210 215 220ctg cac atg gca gcg cag tcc ctg cgt tcg ggc gag tcg tcg ctg gcg720Leu His Met Ala Ala Gln Ser Leu Arg Ser Gly Glu Ser Ser Leu Ala225 230 235 240ctg gtc gga ggc acc aac gtg atg gcc acg ccc act gcc ttc gtg ctg768Leu Val Gly Gly Thr Asn Val Met Ala Thr Pro Thr Ala Phe Val Leu245 250 255acc gcg cgt gca ggg ggc ctg gcg aag gac ggc cgg tgc aag gcg ttc816Thr Ala Arg Ala Gly Gly Leu Ala Lys Asp Gly Arg Cys Lys Ala Phe260 265 270gcg gca tcc gcg gat ggc acg aac tgg gcc gag ggc gtg ggc gtc ctg864Ala Ala Ser Ala Asp Gly Thr Asn Trp Ala Glu Gly Val Gly Val Leu275 280 285ctg ctg gag cgg ctt tcc gac gcc gtc cgc aac ggc cgt gag gtc ctc912Leu Leu Glu Arg Leu Ser Asp Ala Val Arg Asn Gly Arg Glu Val Leu290 295 300ggc gtc gta cgg gcc acc gcg gtg aat cag gat ggc gcg tcc aac gga960Gly Val Val Arg Ala Thr Ala Val Asn Gln Asp Gly Ala Ser Asn Gly305 310 315 320ctc gcc gcg ccc aac ggg ccc tcg cag cag cgg gtg atc cgc cag gca 1008Leu Ala Ala Pro Asn Gly Pro Ser Gln Gln Arg Val Ile Arg Gln Ala325 330 335ctg gcg gcc ggc ggc ctg tcg ccg gcc gac gtc gac atc gtc gag gcg1056Leu Ala Ala Gly Gly Leu Ser Pro Ala Asp Val Asp Ile Val Glu Ala340 345 350cac ggc acc gga acc gcc ctc ggc gac ccc atc gag gca cag gcg ctc1104His Gly Thr Gly Thr Ala Leu Gly Asp Pro Ile Glu Ala Gln Ala Leu355 360 365ctc acc acc tat ggt cgg aac cgt gcc ccc gga ctg ccg ctg tgg ctt1152Leu Thr Thr Tyr Gly Arg Asn Arg Ala Pro Gly Leu Pro Leu Trp Leu370 375 380ggt tcg gtg aag tcg aac ctt gga cac gcg ggc gcc gct gcg ggc gtc1200Gly Ser Val Lys Ser Asn Leu Gly His Ala Gly Ala Ala Ala Gly Val385 390 395 400gcc ggt gtg atc aag atg gtg atg gcg atg cgg cat ggt gtg ctg ccg1248Ala Gly Val Ile Lys Met Val Met Ala Met Arg His Gly Val Leu Pro405 410 415cgg aca ctg cat gtg gac gag ccg acg ccc gag gtc gac tgg tct gcc1296Arg Thr Leu His Val Asp Glu Pro Thr Pro Glu Val Asp Trp Ser Ala420 425 430gga gcg gtc gag ctg ctg acc gag gcg cac gag tgg ccc gag gtc ggc1344Gly Ala Val Glu Leu Leu Thr Glu Ala His Glu Trp Pro Glu Val Gly435 440 445cgt cct cgc cgt gcg ggg gtc tcc ggc ttc ggc gcc agc ggc acc aac1392Arg Pro Arg Arg Ala Gly Val Ser Gly Phe Gly Ala Ser Gly Thr Asn450 455 460gca cac gtc atc ctg gag cag gcg acc gag ccg aca tcc ggg aac ctg1440Ala His Val Ile Leu Glu Gln Ala Thr Glu Pro Thr Ser Gly Asn Leu465 470 475 480ccc gac gag aag gca cgc gtg ctg ggc gac tcg gtt gtg ccg ctg gtc1488Pro Asp Glu Lys Ala Arg Val Leu Gly Asp Ser Val Val Pro Leu Val485 490 495gtc tcg gcc cgt ggc aag gcg ggt ctt gcc ggc cag gct cac cgt ctc1536Val Ser Ala Arg Gly Lys Ala Gly Leu Ala Gly Gln Ala His Arg Leu500 505 510ggc tcg ttc ctg aca cag cgt caa gac acg gac gtg ctc gac atc ggc1584Gly Ser Phe Leu Thr Gln Arg Gln Asp Thr Asp Val Leu Asp Ile Gly515 520 525cag tcg ctg gtg cgg agc cgg ggt cca ctc cag gac cgt gcg gtc gtg1632Gln Ser Leu Val Arg Ser Arg Gly Pro Leu Gln Asp Arg Ala Val Val530 535 540ctc gcc gcg gac cgg gac gag gcg ctg gcc gga ctc gac gct gtg gcc1680Leu Ala Ala Asp Arg Asp Glu Ala Leu Ala Gly Leu Asp Ala Val Ala545 550 555 560cgc gcc gag tcc gcg ccc ggt gtg gtc acg gga ttt gcc gag agc aca1728Arg Ala Glu Ser Ala Pro Gly Val Val Thr Gly Phe Ala Glu Ser Thr565 570 575gtg ggc cgg acc gtc ctc gtg ttc ccc ggc cag ggc aca cag tgg gcg1776Val Gly Arg Thr Val Leu Val Phe Pro Gly Gln Gly Thr Gln Trp Ala580 585 590gga atg gga gcg gaa ctg ctc gaa gcc tca cct gtg ttc gca gcc agg1824Gly Met Gly Ala Glu Leu Leu Glu Ala Ser Pro Val Phe Ala Ala Arg595 600 605atg acc gag tgc gcc gag gtg ctc gac ccg ctg acc ggc tgg tcg ctg1872Met Thr Glu Cys Ala Glu Val Leu Asp Pro Leu Thr Gly Trp Ser Leu610 615 620ctc gat gtg gta cgg cag gtg gag ggc gcc cgg tct ctt gaa gac gtc1920Leu Asp Val Val Arg Gln Val Glu Gly Ala Arg Ser Leu Glu Asp Val625 630 635 640gac gtc ttg cag ccg gtg tcg tgg gca ctg atg gtg tcg ctg gcc gcg1968Asp Val Leu Gln Pro Val Ser Trp Ala Leu Met Val Ser Leu Ala Ala645 650 655ttg tgg gag gcg tgc ggg gtc gtc ccg gac gct gtc gtg ggt cat tcc20l6Leu Trp Glu Ala Cys Gly Val Val Pro Asp Ala Val Val Gly His Ser660 665 670ctg ggc gag atc gcc gct gcc tgc tat gcc ggt gcg ctg tcc ctt ccc2064Leu Gly Glu Ile Ala Ala Ala Cys Tyr Ala Gly Ala Leu Ser Leu Pro675 680 685gac gcc gcc cgc ctc atg gtt cac cgg tcc agg att gcc gaa gcc gag2112Asp Ala Ala Arg Leu Met Val His Arg Ser Arg Ile Ala Glu Ala Glu690 695 700ctg gtg gga cgc gga ggc atg gcg tcc ctc acc gcc gat gtc aag gcc2160Leu Val Gly Arg Gly Gly Met Ala Ser Leu Thr Ala Asp Val Lys Ala705 710 715 720gtc tcc att ctg atc gag gag tgg ccg ggt ctg gag atc gcc gcg gtc2208Val Ser Ile Leu Ile Glu Glu Trp Pro Gly Leu Glu Ile Ala Ala Val725 730 735aac gga ccc gcc tcc gtg gtg gtg acc ggt gaa ctg ccc tcc ctg gaa2256Asn Gly Pro Ala Ser Val Val Val Thr Gly Glu Leu Pro Ser Leu Glu740 745 750gag ctg ctc gcc cga tgc gaa gcc gac ggc atc cgc gcc cgc agg att2304Glu Leu Leu Ala Arg Cys Glu Ala Asp Gly Ile Arg Ala Arg Arg Ile755 760 765cgc ggc atc aac ggc gcc gca cac tcc tca cag atc gac gtg ctg cac2352Arg Gly Ile Asn Gly Ala Ala His Ser Ser Gln Ile Asp Val Leu His770 775 780gac tct ttc gtg gag gcc ctc gcc tcg gtc tcc gcc ggg gct tcg cgc2400Asp Ser Phe Val Glu Ala Leu Ala Ser Val Ser Ala Gly Ala Ser Arg785 790 795 800gta ccg ctg tac tcc acg gtg acc ggg cga ctc cat gac acc acg gag2448Val Pro Leu Tyr Ser Thr Val Thr Gly Arg Leu His Asp Thr Thr Glu805 810 815ttc gac gtc gag cac tgg ttc cgc aac atg cgg cag acc gtg cag ttc2496Phe Asp Val Glu His Trp Phe Arg Asn Met Arg Gln Thr Val Gln Phe820 825 830gac ccg gcc atc cgg tcc ctg gtc ggc gac ggg cac ggc gtg ttc atc2544Asp Pro Ala Ile Arg Ser Leu Val Gly Asp Gly His Gly Val Phe Ile835 840 845gag gtc agt gct cat cct gtg ctg acg tcg agc gtc cag gac gtg ctg2592Glu Val Ser Ala His Pro Val Leu Thr Ser Ser Val Gln Asp Val Leu850 855 860gcg gac ctc gag gcc gga ccg gcc gtc gtc acc ggg acg ctg cgc cgc2640Ala Asp Leu Glu Ala Gly Pro Ala Val Val Thr Gly Thr Leu Arg Arg865 870 875 880gac gac ggc ggc ccg cgc cgg ttc ctc gcc tcg ctg gcc cac ctg tac2688Asp Asp Gly Gly Pro Arg Arg Phe Leu Ala Ser Leu Ala His Leu Tyr885 890 895acc cac ggc gta cgg gtc gac tgg gaa gcc gtc ctc ggt cgc ggc ggg2736Thr His Gly Val Arg Val Asp Trp Glu Ala Val Leu Gly Arg Gly Gly900 905 910gaa cag ccc gta gac ctg ccg acg tac gcc ttt cag cgc cag cgg tac2784Glu Gln Pro Val Asp Leu Pro Thr Tyr Ala Phe Gln Arg Gln Arg Tyr915 920 925tgg ctg gag acg gca gag tcc cgt ggg gac gca ccg ggc ctc ggt ctg2832Trp Leu Glu Thr Ala Glu Ser Arg Gly Asp Ala Pro Gly Leu Gly Leu930 935 940gag gtg gcg aac cat ccc ctg ctc ggc gcg gtt acc gag atc ccc ggc2880Glu Val Ala Asn His Pro Leu Leu Gly Ala Val Thr Glu Ile Pro Gly945 950 955 960tcg gac ggc gtg ctg ttc act tcc cgg ctg tcg ctg cgc aca cac ccc2928Ser Asp Gly Val Leu Phe Thr Ser Arg Leu Ser Leu Arg Thr His Pro965 970 975tgg ctc gcc gac cac gcg ggc gcc gga gtc gtc ctc ctg ccg gga gcg2976Trp Leu Ala Asp His Ala Gly Ala Gly Val Val Leu Leu Pro Gly Ala980 985 990gcc ttc gtg gaa ctc gca gtc cgt gcc gcg gac gag gtc ggc tac ggg 3024Ala Phe Val Glu Leu Ala Val Arg Ala Ala Asp Glu Val Gly Tyr Gly995 1000 1005ctg gtc ggc gaa ctg gtc atc gag cgc ccc ctg gtg ctg ccc gag 3069Leu Val Gly Glu Leu Val Ile Glu Arg Pro Leu Val Leu Pro Glu1010 1015 1020agc ggc ggc gtc cag gta cgc gtg tgg gtc ggc gag ccc gac gag 3114Ser Gly Gly Val Gln Val Arg Val Trp Val Gly Glu Pro Asp Glu1025 1030 1035tcc ggc cac cgt acc gtc cag gtt cac tcc cgc cgg gag gaa gcc 3159Ser Gly His Arg Thr Val Gln Val His Ser Arg Arg Glu Glu Ala1040 1045 1050ggc tcg cga ggg agc tgg acc cgt cat gtc tcc ggg cgg ctg gtg 3204Gly Ser Arg Gly Ser Trp Thr Arg His Val Ser Gly Arg Leu Val1055 1060 1065ccg gag gac ggc cag gcc gag ttc gac ctc acc cag tgg ccg ccg 3249Pro Glu Asp Gly Gln Ala Glu Phe Asp Leu Thr Gln Trp Pro Pro1070 1075 1080ccc ggc gcc acc gcg gtc gac ccg gac gcg ttc gcc cac gcg tac 3294Pro Gly Ala Thr Ala Val Asp Pro Asp Ala Phe Ala His Ala Tyr1085 1090 1095gac cac ttg gca gag gcg gga tac cac tat ggt cca gcc ttt cag3339Asp His Leu Ala Glu Ala Gly Tyr His Tyr Gly Pro Ala Phe Gln1100 1105 1110gga atg cgc gcg gct tgg act cgt ggc gag gag gtg ttc gcc gag3384Gly Met Arg Ala Ala Trp Thr Arg Gly Glu Glu Val Phe Ala Glu1115 1120 1125gtc tca ctg ccg gag tcg gcg ggc aag gcc gat gag tac ggg ttg3429Val Ser Leu Pro Glu Ser Ala Gly Lys Ala Asp Glu Tyr Gly Leu1130 1135 1140cac ccg gcc ctg ctg gac gcg gcc atg cac acc agt ctc ttc cgc3474His Pro Ala Leu Leu Asp Ala Ala Met His Thr Ser Leu Phe Arg1145 1150 1155ccc gat ctg agc gac gag agc ccg aag ctg gct ctg ccg ttc gtc3519Pro Asp Leu Ser Asp Glu Ser Pro Lys Leu Ala Leu Pro Phe Val1160 1165 1170tgg cgc gat gtc agg ctg cac gcc gac gga gcc tcc acg ctg cgg3564Trp Arg Asp Val Arg Leu His Ala Asp Gly Ala Ser Thr Leu Arg1175 1180 1185gtg cac ctc acc ccg ctc gcc ccc gac acg atc cgc ttg cac ctg3609Val His Leu Thr Pro Leu Ala Pro Asp Thr Ile Arg Leu His Leu1190 1195 1200gcc gac act tcc ggc aca ccc gtg gct tcg gtc gac tcg ttg gtc3654Ala Asp Thr Ser Gly Thr Pro Val Ala Ser Val Asp Ser Leu Val1205 1210 1215ctg cgc ccc gtg gtc ccg gaa ctg ctg cgc gtc ggc tca ggc gcg3699Leu Arg Pro Val Val Pro Glu Leu Leu Arg Val Gly Ser Gly Ala1220 1225 1230gcc aag gac cag atg ttc cgg gtg gcc tgg gag ccc atc tcc gtc3744Ala Lys Asp Gln Met Phe Arg Val Ala Trp Glu Pro Ile Ser Val1235 1240 1245agg agc gtg gac gac gag ctg aag gcc gta cgc gtg acg act gcc3789Arg Ser Val Asp Asp Glu Leu Lys Ala Val Arg Val Thr Thr Ala1250 1255 1260gag gac gtc cgt gcc gcg gcc gca acg gcc ccg cgt gtg ctc ctg3834Glu Asp Val Arg Ala Ala Ala Ala Thr Ala Pro Arg Val Leu Leu1265 1270 1275ctc gat gtg gcc ggc gat gga cgt acg gac ccc gac gcg gcc cgg3879Leu Asp Val Ala Gly Asp Gly Arg Thr Asp Pro Asp Ala Ala Arg1280 1285 1290gac ctc agc ggg cgg gtg ctg gag gcc gtc cag gcg tgg ctg gcg3924Asp Leu Ser Gly Arg Val Leu Glu Ala Val Gln Ala Trp Leu Ala1295 1300 1305gag ccc gcc ttc cag gac act gtt ctc ctc gct ctc aca cac tcc3969Glu Pro Ala Phe Gln Asp Thr Val Leu Leu Ala Leu Thr His Ser1310 1315 1320ggg gcg gcc gtc cgg gat ggg gac ccg gtt ccc gat ctc gcc gtt4014Gly Ala Ala Val Arg Asp Gly Asp Pro Val Pro Asp Leu Ala Val1325 1330 1335gcg acg gcc gcc ggc ctg ctg cgt gcg gcg cag tcc gag aac gtg4059Ala Thr Ala Ala Gly Leu Leu Arg Ala Ala Gln Ser Glu Asn Val1340 1345 1350ggc cgc atc atc ctg gtc gac acg gac ggc acg gag gcg tca gcc4104Gly Arg Ile Ile Leu Val Asp Thr Asp Gly Thr Glu Ala Ser Ala1355 1360 1365cgg cgc ctg ccc gat gtg ctc gcg gcc ggg gaa ccg cag gcg gca4149Arg Arg Leu Pro Asp Val Leu Ala Ala Gly Glu Pro Gln Ala Ala1370 1375 1380ctt cgg tcg ggt tcg gtg gcg gtg ccg agg ctc gtc agg gcc tcc4194Leu Arg Ser Gly Ser Val Ala Val Pro Arg Leu Val Arg Ala Ser1385 1390 1395ccc gcc gag gcc cag ggc cgt ccg ctg aac ccc ggg ggt acg gtt4239Pro Ala Glu Ala Gln Gly Arg Pro Leu Asn Pro Gly Gly Thr Val1400 1405 1410ctg atc acc ggc ggt acg ggt tcg ctg ggt cgc ctg gcg gcc ggg4284Leu Ile Thr Gly Gly Thr Gly Ser Leu Gly Arg Leu Ala Ala Gly1415 1420 1425cac ctg gtc acc gag cac aag atc agg agt ctg ctc ctg gtg agc4329His Leu Val Thr Glu His Lys Ile Arg Ser Leu Leu Leu Val Ser1430 1435 1440cgg caa gga ccg gac gca ccg ggc gcg gcc gag ctg gag gcc gaa4374Arg Gln Gly Pro Asp Ala Pro Gly Ala Ala Glu Leu Glu Ala Glu1445 1450 1455ctc acg gaa ctc ggc gcg aac gtc cgg atc gtc gcg tgc gac gtc4419Leu Thr Glu Leu Gly Ala Asn Val Arg Ile Val Ala Cys Asp Val1460 1465 1470tcc gat cgg gac tcc gtg gcc gcg ctg ctg gcc tct gtt ccc cac4464Ser Asp Arg Asp Ser Val Ala Ala Leu Leu Ala Ser Val Pro His1475 1480 1485gac gcc ccg ctc acc ggc gtg atc cac gca gcc ggg gtg ctg gat4509Asp Ala Pro Leu Thr Gly Val Ile His Ala Ala Gly Val Leu Asp1490 1495 1500gac ggt gtg gtc acc tcc ctg acg ccc gaa cgg ctc gac acg gtg4554Asp Gly Val Val Thr Ser Leu Thr Pro Glu Arg Leu Asp Thr Val1505 1510 1515ctc cgt ccc aag gcc gac gcg gca cag atc ctg gac gaa ctc acg4599Leu Arg Pro Lys Ala Asp Ala Ala Gln Ile Leu Asp Glu Leu Thr1520 1525 1530cgc gat ctc gac ctt gcc gtc ttc gtc ctg tac tcc tcc atc gcg4644Arg Asp Leu Asp Leu Ala Val Phe Val Leu Tyr Ser Ser Ile Ala1535 1540 1545ggg atc ttc ggt tca gcg ggc cag agc agc tat gcc gcc gcg aac4689Gly Ile Phe Gly Ser Ala Gly Gln Ser Ser Tyr Ala Ala Ala Asn1550 1555 1560tcg ttc ctc gac gcg ctc gct gaa cgc cgt cgc gct tgc gga ctg4734Ser Phe Leu Asp Ala Leu Ala Glu Arg Arg Arg Ala Cys Gly Leu1565 1570 1575ccg gcg acc tca ctg gtg tgg gga tgg tgg ggc cag gtg tcc ggc4779Pro Ala Thr Ser Leu Val Trp Gly Trp Trp Gly Gln Val Ser Gly1580 1585 1590ata gtg gac aag ctc gcc gag gtc gac ctg aag cgc ttc gac cgg4824Ile Val Asp Lys Leu Ala Glu Val Asp Leu Lys Arg Phe Asp Arg1595 1600 1605ctc aac atg atc gag ttc acc gca caa gag ggc atg gag ctg ttc4869Leu Asn Met Ile Glu Phe Thr Ala Gln Glu Gly Met Glu Leu Phe1610 1615 1620gac ctt gcg ctg tcc gat cgc agc gct gcc ttg gtc ctg gcg aag4914Asp Leu Ala Leu Ser Asp Arg Ser Ala Ala Leu Val Leu Ala Lys1625 1630 1635atg gac ctc aaa gca atg cgg gac cag acc gac tcc gca tct gtc4959Met Asp Leu Lys Ala Met Arg Asp Gln Thr Asp Ser Ala Ser Val1640 1645 1650gcc ccg ctg ctg cgc ggc ctc gtc cgc gtg ggt cgg cgg gcc gcc5004Ala Pro Leu Leu Arg Gly Leu Val Arg Val Gly Arg Arg Ala Ala1655 1660 1665agt gac ggg act rcc ggg ycc gyc ggg ctg gca ggg cgy ktg ycc5049Ser Asp Gly Thr Xaa Gly Xaa Xaa Gly Leu Ala Gly Arg Xaa Xaa1670 1675 1680gag gcg tcc kcc gac cag cgc gga aag atc ttg gcc gac ttg gtc5094Glu Ala Ser Xaa Asp Gln Arg Gly Lys Ile Leu Ala Asp Leu Val1685 1690 1695cag cgc gag gtc tcc gcg atc ctc ggt cac ctg tcg ccg gac cag5139Gln Arg Glu Val Ser Ala Ile Leu Gly His Leu Ser Pro Asp Gln1700 1705 1710atc gga ttg gac ctg tcc ttc ttc gac cte ggg ttc gac tcg ctg5184Ile Gly Leu Asp Leu Ser Phe Phe Asp Leu Gly Phe Asp Ser Leu1715 1720 1725acc gcc gtc gag ctc gcg aac cgg ctg tcg gcg ctg acc ggc ctc5229Thr Ala Val Glu Leu Ala Asn Arg Leu Ser Ala Leu Thr Gly Lcu1730 1735 1740cgt atc ccg tcc acc ttc gcc ttc gac tgc ccc acg gtg gac ctg5274Arg Ile Pro Ser Thr Phe Ala Phe Asp Cys Pro Thr Val Asp Leu1745 1750 1755gct gtc gag gcg ctg ctg gag agc ttc gaa ctc gac gtg gac tag5319Ala Val Glu Ala Leu Leu Glu Ser Phe Glu Leu Asp Val Asp17601765 1770<210>5<211>1772<212>PRT<213>吸水鏈霉菌(Streptomyces hygroscopicus)<220><221>misc_feature<222>(1673)<223>Xaa＝Ala或Thr.<220><221>misc_feature<222>(1675)<223>Xaa或Pro或Ser.<220><221>misc_feature<222>(1676)<223>Xaa＝Ala或Val.<220><221>misc_feature<222>(1682)<223>Xaa＝Val或Leu.<220><221>misc_feature<222>(1683)<223>Xaa或Pro或Ser.<220><221>misc_feature<222>(1687)<223>Xaa＝Ala或Ser.<400>5Val Asp Ala Arg Gln Asn Thr Glu Ile Ala Val Ala Ser Ser Glu Ser1 5 10 15Lys Val Val Glu Ala Leu Arg Ala Ser Leu Met Glu Asn Glu Arg Leu20 25 30Glu Ser Glu Val Gln Ser Ile Arg Asp Ser Leu Thr Glu Pro Ile Ala35 40 45Ile Val Gly Met Ala Cys Arg Phe Pro Gly Gly Val Ser Ser Pro Glu50 55 60Glu Leu Trp Glu Leu Ile Ala Asp Gly Arg Ser Ala Val Glu Ala Phe65 70 75 80Pro Thr Asn Arg Gly Trp Asp Leu Glu Asn Leu Tyr Asp Pro Asp Leu85 90 95Asp Arg Pro Gly Thr Thr Tyr Val Arg Glu Gly Ala Phe Leu His Asp100 105 110Ala Gly Glu Phe Asp Ala Gly Phe Phe Gly Ile Ser Gln Ser Glu Thr115 120 125Met Val Met Asp Pro Gln Gln Arg Leu Met Leu Glu Thr Ser Trp Glu130 135 140Ala Phe Glu Arg Ala Gly Ile Asp Pro Ala Ala Met Arg Gly Lys Asn145 150 155 160Val Gly Val Phe Ala Gly Met Ala Ala Gly Gln Glu Tyr Gly Thr Ala165 170 175Phe His Ser Ile Pro Asp Glu Leu Glu Gly Tyr Val Met Thr Gly Gly180 185 190Leu Ala Ser Val Leu Ser Gly Arg Val Ser Tyr Thr Phe Gly Phe Glu195 200 205Gly Pro Ala Val Thr Ile Asp Thr Ala Cys Ser Ser Ser Leu Val Ala210 215 220Leu His Met Ala Ala Gln Ser Leu Arg Ser Gly Glu Ser Ser Leu Ala225 230 235 240Leu Val Gly Gly Thr Asn Val Met Ala Thr Pro Thr Ala Phe Val Leu245 250 255Thr Ala Arg Ala Gly Gly Leu Ala Lys Asp Gly Arg Cys Lys Ala Phe260 265 270Ala Ala Ser Ala Asp Gly Thr Asn Trp Ala Glu Gly Val Gly Val Leu275 280 285Leu Leu Glu Arg Leu Ser Asp Ala Val Arg Asn Gly Arg Glu Val Leu290 295 300Gly Val Val Arg Ala Thr Ala Val Asn Gln Asp Gly Ala Ser Asn Gly305 310 315 320Leu Ala Ala Pro Asn Gly Pro Ser Gln Gln Arg Val Ile Arg Gln Ala325 330 335Leu Ala Ala Gly Gly Leu Ser Pro Ala Asp Val Asp Ile Val Glu Ala340 345 350His Gly Thr Gly Thr Ala Leu Gly Asp Pro Ile Glu Ala Gln Ala Leu355 360 365Leu Thr Thr Tyr Gly Arg Asn Arg Ala Pro Gly Leu Pro Leu Trp Leu370 375 380Gly Ser Val Lys Ser Asn Leu Gly His Ala Gly Ala Ala Ala Gly Val385 390 395 400Ala Gly Val Ile Lys Met Val Met Ala Met Arg His Gly Val Leu Pro405 410 415Arg Thr Leu His Val Asp Glu Pro Thr Pro Glu Val Asp Trp Ser Ala420 425 430Gly Ala Val Glu Leu Leu Thr Glu Ala His Glu Trp Pro Glu Val Gly435 440 445Arg Pro Arg Arg Ala Gly Val Ser Gly Phe Gly Ala Ser Gly Thr Asn450 455 460Ala His Val Ile Leu Glu Gln Ala Thr Glu Pro Thr Ser Gly Asn Leu465 470 475 480Pro Asp Glu Lys Ala Arg Val Leu Gly Asp Ser Val Val Pro Leu Val485 490 495Val Ser Ala Arg Gly Lys Ala Gly Leu Ala Gly Gln Ala His Arg Leu500 505 510Gly Ser Phe Leu Thr Gln Arg Gln Asp Thr Asp Val Leu Asp Ile Gly515 520 525Gln Ser Leu Val Arg Ser Arg Gly Pro Leu Gln Asp Arg Ala Val Val530 535 540Leu Ala Ala Asp Arg Asp Glu Ala Leu Ala Gly Leu Asp Ala Val Ala545 550 555 560Arg Ala Glu Ser Ala Pro Gly Val Val Thr Gly Phe Ala Glu Ser Thr565 570 575Val Gly Arg Thr Val Leu Val Phe Pro Gly Gln Gly Thr Gln Trp Ala580 585 590Gly Met Gly Ala Glu Leu Leu Glu Ala Ser Pro Val Phe Ala Ala Arg595 600 605Met Thr Glu Cys Ala Glu Val Leu Asp Pro Leu Thr Gly Trp Ser Leu6l0 615 620Leu Asp Val Val Arg Gln Val Glu Gly Ala Arg Ser Leu Glu Asp Val625 630 635 640Asp Val Leu Gln Pro Val Ser Trp Ala Leu Met Val Ser Leu Ala Ala645 650 655Leu Trp Glu Ala Cys Gly Val Val Pro Asp Ala Val Val Gly His Ser660 665 670Leu Gly Glu Ile Ala Ala Ala Cys Tyr Ala Gly Ala Leu Ser Leu Pro675 680 685Asp Ala Ala Arg Leu Met Val His Arg Ser Arg Ile Ala Glu Ala Glu690 695 700Leu Val Gly Arg Gly Gly Met Ala Ser Leu Thr Ala Asp Val Lys Ala705 710 715 720Val Ser Ile Leu Ile Glu Glu Trp Pro Gly Leu Glu Ile Ala Ala Val725 730 735Asn Gly Pro Ala Ser Val Val Val Thr Gly Glu Leu Pro Ser Leu Glu740 745 750Glu Leu Leu Ala Arg Cys Glu Ala Asp Gly Ile Arg Ala Arg Arg Ile755 760 765Arg Gly Ile Asn Gly Ala Ala His Ser Ser Gln Ile Asp Val Leu His770 775 780Asp Ser Phe Val Glu Ala Leu Ala Ser Val Ser Ala Gly Ala Ser Arg785 790 795 800Val Pro Leu Tyr Ser Thr Val Thr Gly Arg Leu His Asp Thr Thr Glu805 810 815Phe Asp Val Glu His Trp Phe Arg Asn Met Arg Gln Thr Val Gln Phe820 825 830Asp Pro Ala Ile Arg Ser Leu Val Gly Asp Gly His Gly Val Phe Ile835 840 845Glu Val Ser Ala His Pro Val Leu Thr Ser Ser Val Gln Asp Val Leu850 855 860Ala Asp Leu Glu Ala Gly Pro Ala Val Val Thr Gly Thr Leu Arg Arg865 870 875 880Asp Asp Gly Gly Pro Arg Arg Phe Leu Ala Ser Leu Ala His Leu Tyr885 890 895Thr His Gly Val Arg Val Asp Trp Glu Ala Val Leu Gly Arg Gly Gly900 905 910Glu Gln Pro Val Asp Leu Pro Thr Tyr Ala Phe Gln Arg Gln Arg Tyr915 920 925Trp Leu Glu Thr Ala Glu Ser Arg Gly Asp Ala Pro Gly Leu Gly Leu930 935 940Glu Val Ala Asn His Pro Leu Leu Gly Ala Val Thr Glu Ile Pro Gly945 950 955 960Ser Asp Gly Val Leu Phe Thr Ser Arg Leu Ser Leu Arg Thr His Pro965 970 975Trp leu Ala Asp His Ala Gly Ala Gly Val Val Leu Leu Pro Gly Ala980 985 990Ala Phe Val Glu Leu Ala Val Arg Ala Ala Asp Glu Val Gly Tyr Gly995 1000 1005Leu Val Gly Glu Leu Val Ile Glu Arg Pro Leu Val Leu Pro Glu1010 1015 1020Ser Gly Gly Val Gln Val Arg Val Trp Val Gly Glu Pro Asp Glu1025 1030 1035Ser Gly His Arg Thr Val Gln Val His Ser Arg Arg Glu Glu Ala1040 1045 1050Gly Ser Arg Gly Ser Trp Thr Arg His Val Ser Gly Arg Leu Val1055 1060 1065Pro Glu Asp Gly Gln Ala Glu Phe Asp Leu Thr Gln Trp Pro Pro1070 1075 1080Pro Gly Ala Thr Ala Val Asp Pro Asp Ala Phe Ala His Ala Tyr1085 1090 1095Asp His Leu Ala Glu Ala Gly Tyr His Tyr Gly Pro Ala Phe Gln1100 1105 1110Gly Met Arg Ala Ala Trp Thr Arg Gly Glu Glu Val Phe Ala Glu1115 1120 1125Val Ser Leu Pro Glu Ser Ala Gly Lys Ala Asp Glu Tyr Gly Leul130 1135 1140His Pro Ala Leu Leu Asp Ala Ala Met His Thr Ser Leu Phe Arg1145 1150 1155Pro Asp Leu Ser Asp Glu Ser Pro Lys Leu Ala Leu Pro Phe Val1160 1165 1170Trp Arg Asp Val Arg Leu His Ala Asp Gly Ala Ser Thr Leu Arg1175 1180 1185Val His Leu Thr Pro Leu Ala Pro Asp Thr Ile Arg Leu His Leu1190 1195 1200Ala Asp Thr Ser Gly Thr Pro Val Ala Ser Val Asp Ser Leu Val1205 1210 1215Leu Arg Pro Val Val Pro Glu Leu Leu Arg Val Gly Ser Gly Ala1220 1225 1230Ala Lys Asp Gln Met Phe Arg Val Ala Trp Glu Pro Ile Ser Val1235 1240 1245Arg Ser Val Asp Asp Glu Leu Lys Ala Val Arg Val Thr Thr Ala1250 1255 1260Glu Asp Val Arg Ala Ala Ala Ala Thr Ala Pro Arg Val Leu Leu1265 1270 1275Leu Asp Val Ala Gly Asp Gly Arg Thr Asp Pro Asp Ala Ala Arg1280 1285 1290Asp Leu Ser Gly Arg Val Leu Glu Ala Val Gln Ala Trp Leu Ala1295 1300 1305Glu Pro Ala Phe Gln Asp Thr Val Leu Leu Ala Leu Thr His Ser1310 1315 1320Gly Ala Ala Val Arg Asp Gly Asp Pro Val Pro Asp Leu Ala Val1325 1330 1335Ala Thr Ala Ala Gly Leu Leu Arg Ala Ala Gln Ser Glu Asn Val1340 1345 1350Gly Arg Ile Ile Leu Val Asp Thr Asp Gly Thr Glu Ala Ser Ala1355 1360 1365Arg Arg Leu Pro Asp Val Leu Ala Ala Gly Glu Pro Gln Ala Ala1370 1375 1380Leu Arg Ser Gly Ser Val Ala Val Pro Arg Leu Val Arg Ala Ser1385 1390 1395Pro Ala Glu Ala Gln Gly Arg Pro Leu Asn Pro Gly Gly Thr Val1400 1405 1410Leu Ile Thr Gly Gly Thr Gly Ser Leu Gly Arg Leu Ala Ala Gly1415 1420 1425His Leu Val Thr Glu His Lys Ile Arg Ser Leu Leu Leu Val Ser1430 1435 1440Arg Gln Gly Pro Asp Ala Pro Gly Ala Ala Glu Leu Glu Ala Glu1445 1450 1455Leu Thr Glu Leu Gly Ala Asn Val Arg Ile Val Ala Cys Asp Val1460 1465 1470Ser Asp Arg Asp Ser Val Ala Ala Leu Leu Ala Ser Val Pro His1475 1480 1485Asp Ala Pro Leu Thr Gly Val Ile His Ala Ala Gly Val Leu Asp1490 1495 1500Asp Gly Val Val Thr Ser Leu Thr Pro Glu Arg Leu Asp Thr Val1505 1510 1515Leu Arg Pro Lys Ala Asp Ala Ala Gln Ile Leu Asp Glu Leu Thr1520 1525 1530Arg Asp Leu Asp Leu Ala Val Phe Val Leu Tyr Ser Ser Ile Ala1535 1540 1545Gly Ile Phe Gly Ser Ala Gly Gln Ser Ser Tyr Ala Ala Ala Asn1550 1555 1560Ser Phe Leu Asp Ala Leu Ala Glu Arg Arg Arg Ala Cys Gly Leu1565 1570 1575Pro Ala Thr Ser Leu Val Trp Gly Trp Trp Gly Gln Val Ser Gly1580 1585 1590Ile Val Asp Lys Leu Ala Glu Val Asp Leu Lys Arg Phe Asp Arg1595 1600 1605Leu Asn Met Ile Glu Phe Thr Ala Gln Glu Gly Met Glu Leu Phe1610 1615 1620Asp Leu Ala Leu Ser Asp Arg Ser Ala Ala Leu Val Leu Ala Lys1625 1630 1635Met Asp Leu Lys Ala Met Arg Asp Gln Thr Asp Ser Ala Ser Val1640 1645 1650Ala Pro Leu Leu Arg Gly Leu Val Arg Val Gly Arg Arg Ala Ala1655 1660 1665Ser Asp Gly Thr Xaa Gly Xaa Xaa Gly Leu Ala Gly Arg Xaa Xaa1670 1675 1680Glu Ala Ser Xaa Asp Gln Arg Gly Lys Ile Leu Ala Asp Leu Val1685 1690 1695Gln Arg Glu Val Ser Ala Ile Leu Gly His Leu Ser Pro Asp Gln1700 1705 1710Ile Gly Leu Asp Leu Ser Phe Phe Asp Leu Gly Phe Asp Ser Leu1715 1720 1725Thr Ala Val Glu Leu Ala Asn Arg Leu Ser Ala Leu Thr Gly Leu1730 1735 1740Arg Ile Pro Ser Thr Phe Ala Phe Asp Cys Pro Thr Val Asp Leu1745 1750 1755Ala Val Glu Ala Leu Leu Glu Ser Phe Glu Leu Asp Val Asp1760 1765 1770<210>6<211>2325<212>DNA<213>吸水鏈霉菌(Streptomyces hygroscopicus)<220><221>CDS<222>(1)..(2325)<223>shnC<220><221>misc_feature<222>(2210)<223>n＝a或g或t或c<400>6atg gcc gag ctc ttc gtc cgg ggt gtt ccc gtc gac tgg acc aag ttc 48Met Ala Glu Leu Phe Val Arg Gly Val Pro Val Asp Trp Thr Lys Phe1 5 10 15ctc atg gcc ggg gcc ggg cac gtc gac ctt ccg acg tac gcc ttc gac 96Leu Met Ala Gly Ala Gly His Val Asp Leu Pro Thr Tyr Ala Phe Asp20 25 30cgg cgc cac tac tgg ttg cag gat gct gcg aca gcc gac gac agc ggc144Arg Arg His Tyr Trp Leu Gln Asp Ala Ala Thr Ala Asp Asp Ser Gly35 40 45gcc tcc gac aac gac gcc gac gcg gac ttc tgg agc gcc gtc gag cag192Ala Ser Asp Asn Asp Ala Asp Ala Asp Phe Trp Ser Ala Val Glu Gln50 55 60acc gac gcg gac tcg ctc gcc ggg ctc ctc gcc ccg gac tcc gcc ggt240Thr Asp Ala Asp Ser Leu Ala Gly Leu Leu Ala Pro Asp Ser Ala Gly65 70 75 80ctg cgc gac gcc ttg cgc acc gtc gtg ccg gcg ctt gcg gac tgg cgc288Leu Arg Asp Ala Leu Arg Thr Val Val Pro Ala Leu Ala Asp Trp Arg85 90 95ggc agg agc cgg cgg cgc tcc agc gct gaa cgc ctc cgc tac gcc gtc336Gly Arg Ser Arg Arg Arg Ser Ser Ala Glu Arg Leu Arg Tyr Ala Val100 105 110acc tgg cgg ccc ctg gac cgt gag gtg tca agg gtc ccc gcg ggc cgc384Thr Trp Arg Pro Leu Asp Arg Glu Val Ser Arg Val Pro Ala Gly Arg115 120 125tgg ctc gcc gtc ctg ccg ccg gga tgc ccg gcc gaa acc gtg acc ggc432Trp Leu Ala Val Leu Pro Pro Gly Cys Pro Ala Glu Thr Val Thr Gly130 135 140tcc cgg gtg gcc gag ctc atc gcg gag ctc ggt gcc cag gga ctc gac480Ser Arg Val Ala Glu Leu Ile Ala Glu Leu Gly Ala Gln Gly Leu Asp145 150 155 160gtg gtg ccc ttc gag acc gtt ccc tcc gcc ttc acc cgc acc gga ctc528Val Val Pro Phe Glu Thr Val Pro Ser Ala Phe Thr Arg Thr Gly Leu165 170 175acc gcg cgc ctg agc gac atc cgg gcc gag tac cag ccc gcg gga gtc576Thr Ala Arg Leu Ser Asp Ile Arg Ala Glu Tyr Gln Pro Ala Gly Val180 185 190ctc tcc ctg ctc gcc ctc gac ggc gag cag gac gtc atc gac acc gtc624Leu Ser Leu Leu Ala Leu Asp Gly Glu Gln Asp Val Ile Asp Thr Val195 200 205gcc agg acc ctc gcg ctg gtt cag gcg ctg ggg gac gcg ggc gtg aac672Ala Arg Thr Leu Ala Leu Val Gln Ala Leu Gly Asp Ala Gly Val Asn210 215 220ggg ccg ctg tgg tgt ctg acc cgg ggc gcg gtg aac acc gga att cag720Gly Pro Leu Trp Cys Leu Thr Arg Gly Ala Val Asn Thr Gly Ile Gln225 230 235 240gac acg gcc ggc gag ccc ggc gac gcc gcg atc tgg ggg ctg ggc cgc768Asp Thr Ala Gly Glu Pro Gly Asp Ala Ala Ile Trp Gly Leu Gly Arg245 250 255gcc gcg gct ctc gag cac ccc gac cgg tgg ggc ggc ctg atc gac ctg816Ala Ala Ala Leu Glu His Pro Asp Arg Trp Gly Gly Leu Ile Asp Leu260 265 270ccg gcg acc gcc gac gcc cac acc gcg cag tac ctc gtg ggc gcg ctg864Pro Ala Thr Ala Asp Ala His Thr Ala Gln Tyr Leu Val Gly Ala Leu275 280 285aac ggc acc gcg ggc gac cag ctc gcc gta cgc cgc ccc ggc ctc tac912Asn Gly Thr Ala Gly Asp Gln Leu Ala Val Arg Arg Pro Gly Leu Tyr290 295 300agc cgg cgg ctc gta cgc aag ccc gcg tct cag acc ccc gcc gac ggc960Ser Arg Arg Leu Val Arg Lys Pro Ala Ser Gln Thr Pro Ala Asp Gly305 310 315 320ggc tgg cgg ccc cac ggc aca gtc ctc gtg acc ggc ggc gcc gag gcc 1008Gly Trp Arg Pro His Gly Thr Val Leu Val Thr Gly Gly Ala Glu Ala325 330 335ctc ggc atc cat gcc tcg ctc tgg ctc gcc cgg tcc ggc gcg cgc cgt 1056Leu Gly Ile His Ala Ser Leu Trp Leu Ala Arg Ser Gly Ala Arg Arg340345 350ctc atc gtc aca acc acg gct cag gcc ccc gcc gac gcc gtc acc gag 1104Leu Ile Val Thr Thr Thr Ala Gln Ala Pro Ala Asp Ala Val Thr Glu355 360 365ttg cag ggc aag ctc gcg gcc gcc ggg gtg gag acg aca gtc gtc tca 1152Leu Gln Gly Lys Leu Ala Ala Ala Gly Val Glu Thr Thr Val Val Ser370 375 380tgt gcc gac gcc gac cgt gag acg ctc gcc cgg ctc atc gcc gag acc 1200Cys Ala Asp Ala Asp Arg Glu Thr Leu Ala Arg Leu Ile Ala Glu Thr385 390 395 400ccg cgg gaa cag ccg ctg acc gcc gtc gtg cac gcc gcc gac gct cca1248Pro Arg Glu Gln Pro Leu Thr Ala Val Val His Ala Ala Asp Ala Pro405 410 415tgg acc agt gcc gtc gcc gac acc ggc cac gcc gac ctc acc gag gtc1296Trp Thr Ser Ala Val Ala Asp Thr Gly His Ala Asp Leu Thr Glu Val420 425 430ttc gcg ggc aag gtc gac acc gct gtg tgg ctc gac gaa ctg ttc acc1344Phe Ala Gly Lys Val Asp Thr Ala Val Trp Leu Asp Glu Leu Phe Thr435 440 445ggc acc gac gcc gcc ccg ctc gac gcc ttc gtg gtc ttc tcc tcg atc1392Gly Thr Asp Ala Ala Pro Leu Asp Ala Phe Val Val Phe Ser Ser Ile450 455 460gcc ggc atc tgg ggc ggc ggt ggc cag ggt gtc tcc ggc gcg gcc ggc1440Ala Gly Ile Trp Gly Gly Gly Gly Gln Gly Val Ser Gly Ala Ala Gly465 470 475 480gcg gtc ctg gac gcc ctt gtc gat cgg cgc cgc ggc cgg gga ctc gcg1488Ala Val Leu Asp Ala Leu Val Asp Arg Arg Arg Gly Arg Gly Leu Ala485 490 495gcc acc tcg atc gcc tgg gga gcc ctc gac ggg atc ggc ctc ggc atg1536Ala Thr Ser Ile Ala Trp Gly Ala Leu Asp Gly Ile Gly Leu Gly Met500 505 510gat gag gcg gcc gcc gcg cag ctg cgc cgc cgc ggt gtc ctg ccg atg1584Asp Glu Ala Ala Ala Ala Gln Leu Arg Arg Arg Gly Val Leu Pro Met515 520 525gcc cat cag gtc gcc gtg acc gcg ttc gaa cag gcc gcg gag gca cgg1632Ala His Gln Val Ala Val Thr Ala Phe Glu Gln Ala Ala Glu Ala Arg530 535 540gag aag gcc gtg acg gtt gcc gac atg gat tgg gaa gcg ttc atc ccg1680Glu Lys Ala Val Thr Val Ala Asp Met Asp Trp Glu Ala Phe Ile Pro545 550 555 560gcg ttc acc tcc gca cga gtc agc ccg ctc ttc gcc gac ttg ccc gaa1728Ala Phe Thr Ser Ala Arg Val Ser Pro Leu Phe Ala Asp Leu Pro Glu565 570 575gcc gcg gcc gca ctg cgc tcc tcc caa ccc gat gcc gag aac ggc gac1776Ala Ala Ala Ala Leu Arg Ser Ser Gln Pro Asp Ala Glu Asn Gly Asp580 585 590atc acc tca tcc ctg gtc gac tct ctg cgg gac gtc ccc cag gcc gaa1824Ile Thr Ser Ser Leu Val Asp Ser Leu Arg Asp Val Pro Gln Ala Glu595 600 605cag aac cgt ctc ctg ctc cgg ctg gtc tgt ggg cag gcc gcg acc gtc1872Gln Asn Arg Leu Leu Leu Arg Leu Val Cys Gly Gln Ala Ala Thr Val610 615 620ctc gga cac agc agt ggg gag agc atc ggt ccg ctc cag tcc ttc cag1920Leu Gly His Ser Ser Gly Glu Ser Ile Gly Pro Leu Gln Ser Phe Gln625 630 635 640gag gtc ggc ttc gac tcg ctc ggc gcc gtc aac ctc cgc aac agc ctg1968Glu Val Gly Phe Asp Ser Leu Gly Ala Val Asn Leu Arg Asn Ser Leu645 650 655cac gtc gcc acc ggt cta cga ctg ccc gcg aca ctc gtc ttc gac tac2016His Val Ala Thr Gly Leu Arg Leu Pro Ala Thr Leu Val Phe Asp Tyr660 665 670ccg acc ccg gac gcc gtc gtc ggc ttc ctg cgc tcc gag ctg ctg acg2064Pro Thr Pro Asp Ala Val Val Gly Phe Leu Arg Ser Glu Leu Leu Thr675680 685gaa acg agc gac gac ctg gaa ggg cgg gag gac gac ctg cga cgc gtg2112Glu Thr Ser Asp Asp Leu Glu Gly Arg Glu Asp Asp Leu Arg Arg Val690 695 700ctc gca cag gtc ccg ctc tcc cgc ctt cgg gag gcc ggc ctt ctc gac2160Leu Ala Gln Val Pro Leu Ser Arg Leu Arg Glu Ala Gly Leu Leu Asp705 710 715 720acg ctg ctc agc ctg ggc gac tcc gtg gac ggc tcc gtc ccc gag gcg2208Thr Leu Leu Ser Leu Gly Asp Ser Val Asp Gly Ser Val Pro Glu Ala725 730 735gng gcg ccc gag ccg gcc ccg gcg gcg ccc gcc gcc gag gac gca gcc2256Xaa Ala Pro Glu Pro Ala Pro Ala Ala Pro Ala Ala Glu Asp Ala Ala740 745 750cgt gat cga cgt gat gga cgt cgc gga cct cgt aaa gcg cgc tct ggg2304Arg Asp Arg Arg Asp Gly Arg Arg Gly Pro Arg Lys Ala Arg Ser Gly755 760 765cag caa ccc caa ctg act taa2325Gln Gln Pro Gln Leu Thr770<210>7<211>774<212>PRT<213>吸水鏈霉菌(Streptomyces hygroscopicus)<220><221>misc_feature<222>(737)<223>Xaa＝Glu或Gly或Ala或Val.<400>7Met Ala Glu Leu Phe Val Arg Gly Val Pro Val Asp Trp Thr Lys Phe1 5 10 15Leu Met Ala Gly Ala Gly His Val Asp Leu Pro Thr Tyr Ala Phe Asp20 25 30Arg Arg His Tyr Trp Leu Gln Asp Ala Ala Thr Ala Asp Asp Ser Gly35 40 45Ala Ser Asp Asn Asp Ala Asp Ala Asp Phe Trp Ser Ala Val Glu Gln50 55 60Thr Asp Ala Asp Ser Leu Ala Gly Leu Leu Ala Pro Asp Ser Ala Gly65 70 75 80Leu Arg Asp Ala Leu Arg Thr Val Val Pro Ala Leu Ala Asp Trp Arg85 90 95Gly Arg Ser Arg Arg Arg Ser Ser Ala Glu Arg Leu Arg Tyr Ala Val
100 105 110Thr Trp Arg Pro Leu Asp Arg Glu Val Ser Arg Val Pro Ala Gly Arg115 120 125Trp Leu Ala Val Leu Pro Pro Gly Cys Pro Ala Glu Thr Val Thr Gly130 135 140Ser Arg Val Ala Glu Leu Ile Ala Glu Leu Gly Ala Gln Gly Leu Asp145 150 155 160Val Val Pro Phe Glu Thr Val Pro Ser Ala Phe Thr Arg Thr Gly Leu165 170 175Thr Ala Arg Leu Ser Asp Ile Arg Ala Glu Tyr Gln Pro Ala Gly Val180 185 190Leu Ser Leu Leu Ala Leu Asp Gly Glu Gln Asp Val Ile Asp Thr Val195 200 205Ala Arg Thr Leu Ala Leu Val Gln Ala Leu Gly Asp Ala Gly Val Asn2l0 215 220Gly Pro Leu Trp Cys Leu Thr Arg Gly Ala Val Asn Thr Gly Ile Gln225 230 235 240Asp Thr Ala Gly Glu Pro Gly Asp Ala Ala Ile Trp Gly Leu Gly Arg245 250 255Ala Ala Ala Leu Glu His Pro Asp Arg Trp Gly Gly Leu Ile Asp Leu260 265 270Pro Ala Thr Ala Asp Ala His Thr Ala Gln Tyr Leu Val Gly Ala Leu275 280 285Asn Gly Thr Ala Gly Asp Gln Leu Ala Val Arg Arg Pro Gly Leu Tyr290 295 300Ser Arg Arg Leu Val Arg Lys Pro Ala Ser Gln Thr Pro Ala Asp Gly305 310 315 320Gly Trp Arg Pro His Gly Thr Val Leu Val Thr Gly Gly Ala Glu Ala325 330 335Leu Gly Ile His Ala Ser Leu Trp Leu Ala Arg Ser Gly Ala Arg Arg340 345 350Leu Ile Val Thr Thr Thr Ala Gln Ala Pro Ala Asp Ala Val Thr Glu355 360 365Leu Gln Gly Lys Leu Ala Ala Ala Gly Val Glu Thr Thr Val Val Ser370 375 380Cys Ala Asp Ala Asp Arg Glu Thr Leu Ala Arg Leu Ile Ala Glu Thr385 390 395 400Pro Arg Glu Gln Pro Leu Thr Ala Val Val His Ala Ala Asp Ala Pro405 410 415Trp Thr Ser Ala Val Ala Asp Thr Gly His Ala Asp Leu Thr Glu Val420 425 430Phe Ala Gly Lys Val Asp Thr Ala Val Trp Leu Asp Glu Leu Phe Thr435 440 445Gly Thr Asp Ala Ala Pro Leu Asp Ala Phe Val Val Phe Ser Ser Ile450 455 460Ala Gly Ile Trp Gly Gly Gly Gly Gln Gly Val Ser Gly Ala Ala Gly465 470 475 480Ala Val Leu Asp Ala Leu Val Asp Arg Arg Arg Gly Arg Gly Leu Ala
485 490 495Ala Thr Ser Ile Ala Trp Gly Ala Leu Asp Gly Ile Gly Leu Gly Met500 505 510Asp Glu Ala Ala Ala Ala Gln Leu Arg Arg Arg Gly Val Leu Pro Met515 520 525Ala His Gln Val Ala Val Thr Ala Phe Glu Gln Ala Ala Glu Ala Arg530 535 540Glu Lys Ala Val Thr Val Ala Asp Met Asp Trp Glu Ala Phe Ile Pro545 550 555 560Ala Phe Thr Ser Ala Arg Val Ser Pro Leu Phe Ala Asp Leu Pro Glu565 570 575Ala Ala Ala Ala Leu Arg Ser Ser Gln Pro Asp Ala Glu Asn Gly Asp580 585 590Ile Thr Ser Ser Leu Val Asp Ser Leu Arg Asp Val Pro Gln Ala Glu595 600605Gln Asn Arg Leu Leu Leu Arg Leu Val Cys Gly Gln Ala Ala Thr Val610 615 620Leu Gly His Ser Ser Gly Glu Ser Ile Gly Pro Leu Gln Ser Phe Gln625 630 635 640Glu Val Gly Phe Asp Ser Leu Gly Ala Val Asn Leu Arg Asn Ser Leu645 650 655His Val Ala Thr Gly Leu Arg Leu Pro Ala Thr Leu Val Phe Asp Tyr660 665 670Pro Thr Pro Asp Ala Val Val Gly Phe Leu Arg Ser Glu Leu Leu Thr675 680 685Glu Thr Ser Asp Asp Leu Glu Gly Arg Glu Asp Asp Leu Arg Arg Val690 695 700Leu Ala Gln Val Pro Leu Ser Arg Leu Arg Glu Ala Gly Leu Leu Asp705 710 715 720Thr Leu Leu Ser Leu Gly Asp Ser Val Asp Gly Ser Val Pro Glu Ala725 730 735Xaa Ala Pro Glu Pro Ala Pro Ala Ala Pro Ala Ala Glu Asp Ala Ala740 745 750Arg Asp Arg Arg Asp Gly Arg Arg Gly Pro Arg Lys Ala Arg Ser Gly755 760 765Gln Gln Pro Gln Leu Thr770<210>8<211>2976<212>DNA<213>吸水鏈霉菌(Streptomyces hygroscopicus)<220><221>CDS<222>(1)..(2976)<223>shnD<220><221>misc_feature<222>(123，2012)<223>k＝g或t；r＝a或g<400>8atg acg gca ccg gac gag cag atc gtc gac gca ctg cgt gcc tcg ctc 48Met Thr Ala Pro Asp Glu Gln Ile Val Asp Ala Leu Arg Ala Ser Leu1 5 10 15aag gag aac atg cgg ctc caa cag gag aac cag cgt ctc tcc gag tcc 96Lys Glu Asn Met Arg Leu Gln Gln Glu Asn Gln Arg Leu Ser Glu Ser20 25 30tcg gcc gag ccc atc gcg atc gtg tcr atg gct tgt cgg tac gcg ggc144Ser Ala Glu Pro Ile Ala Ile Val Xaa Met Ala Cys Arg Tyr Ala Gly35 40 45ggc ata cgc aac ccc gag gac ctc tgg cgg gtg gtg aac gac ggc acc192Gly Ile Arg Asn Pro Glu Asp Leu Trp Arg Val Val Asn Asp Gly Thr50 55 60gac gtc tac acc tcc ttc ccc gag aac cgc ggc tgg gac ctg gag ggc240Asp Val Tyr Thr Ser Phe Pro Glu Asn Arg Gly Trp Asp Leu Glu Gly65 70 75 80atc tac cac ccc gac ccg gac aac ccc ggc acg acg tac gtc cgc gag288Ile Tyr His Pro Asp Pro Asp Asn Pro Gly Thr Thr Tyr Val Arg Glu85 90 95ggt gcg ttc ctg cac gac gcc aac ctg ttc gac gcc ggg ttg ttc ggg336Gly Ala Phe Leu His Asp Ala Asn Leu Phe Asp Ala Gly Leu Phe Gly100 105 110atc tcg ccg ctt gag gcg cta gcg atg gaa cct caa cag cgg cag ctt384Ile Ser Pro Leu Glu Ala Leu Ala Met Glu Pro Gln Gln Arg Gln Leu115 120 125ctc gag atc tgc tgg gag gcc ctc gaa cga gcc ggc atc gac ccg cac432Leu Glu Ile Cys Trp Glu Ala Leu Glu Arg Ala Gly Ile Asp Pro His130 135 140tcc gta cgc ggc gcc gac atc ggc gta tac gcc ggt ctg gtc cac cag480Ser Val Arg Gly Ala Asp Ile Gly Val Tyr Ala Gly Leu Val His Gln145 150 155 160gac tac gcg ccc gac ctc agc ggc ctc gaa ggc tac ctc agc ctg gag528Asp Tyr Ala Pro Asp Leu Ser Gly Leu Glu Gly Tyr Leu Ser Leu Glu165 170 175cgt gct ctg ggc agc gcg ggc ggc atc gcc tcg gga cgg gtc gcc tac576Arg Ala Leu Gly Ser Ala Gly Gly Ile Ala Ser Gly Arg Val Ala Tyr180 185 190aca ctc ggc ctc gaa ggc ccg gcc gtc acc gtc gac acc atg tgc tcc624Thr Leu Gly Leu Glu Gly Pro Ala Val Thr Val Asp Thr Met Cys Ser195 200 205tcc acc ctg gtc gcc gtg cac gtg gcc aca cag gcg ctt cgg cgc ggt672Ser Thr Leu Val Ala Val His Val Ala Thr Gln Ala Leu Arg Arg Gly210 215 220gag tgc gcc atg gcc ctg gcc ggc ggc gcg acc gtc atg tcg acc ccc720Glu Cys Ala Met Ala Leu Ala Gly Gly Ala Thr Val Met Ser Thr Pro225 230 235 240gga ggg ttc atc ggc ttc gcc cgg cag cgc gcc ctc gcc ttc gac ggc768Gly Gly Phe Ile Gly Phe Ala Arg Gln Arg Ala Leu Ala Phe Asp Gly245 250 255cgc tgc aag tcg tac ggg gcc gcc gcg gac ggc tcc agc tgg gcc gag816Arg Cys Lys Ser Tyr Gly Ala Ala Ala Asp Gly Ser Ser Trp Ala Glu260 265 270ggc gcc ggt gtc gtc ctc ctc gag cgg ctg tcg gac gcg cgc cgc aac864Gly Ala Gly Val Val Leu Leu Glu Arg Leu Ser Asp Ala Arg Arg Asn275 280 285gga cac cgg gtc ctc gcg gtg atc cgc ggc tcc gcc ctc aac cag gac912Gly His Arg Val Leu Ala Val Ile Arg Gly Ser Ala Leu Asn Gln Asp290 295 300ggc gcc tcc aac ggt ctg acg gcg ccc aat ggc ccg gcg cag cgg cgc960Gly Ala Ser Asn Gly Leu Thr Ala Pro Asn Gly Pro Ala Gln Arg Arg305 310 315 320gtc atc cgc aag gcg ctg gag aac gcc ggc ctc acc aca gcc gac atc 1008Val Ile Arg Lys Ala Leu Glu Asn Ala Gly Leu Thr Thr Ala Asp Ile325 330 335gac atg gtc gag ggc cac ggc acc ggc acc gtt ctc ggc gac ccg atc 1056Asp Met Val Glu Gly His Gly Thr Gly Thr Val Leu Gly Asp Pro Ile340 345350gag gcc cag gcc ctg atc gcc acg tac ggc cag gac cgg ccc gag ggc 1104Glu Ala Gln Ala Leu Ile Ala Thr Tyr Gly Gln Asp Arg Pro Glu Gly355 360 365cgg ccg cta tgg ctc ggc tcg gtc aag tcg gtg atc gga cac acc cag 1152Arg Pro Leu Trp Leu Gly Ser Val Lys Ser Val Ile Gly His Thr Gln370 375 380agc ggc tcc ggc gtg gcc gga ctg atc aac gcg gtg cag gcg ctc agg 1200Ser Gly Ser Gly Val Ala Gly Leu Ile Asn Ala Val Gln Ala Leu Arg385 390 395 400cac ggc gtc atg ccc gcc acc cgg cac gtc gac gcc ccc aac ccg cag 1248His Gly Val Met Pro Ala Thr Arg His Val Asp Ala Pro Asn Pro Gln405 410 415gtg gac tgg tcg gcg ggt gcg gtg gag ctg ctg acc gag gcc cgc gcg 1296Val Asp Trp Ser Ala Gly Ala Val Glu Leu Leu Thr Glu Ala Arg Ala420 425 430tgg ccg gag ctg ggc cgg cca cgc cgg gcc ggt gtg tcc tcg ttc ggc 1344Trp Pro Glu Leu Gly Arg Pro Arg Arg Ala Gly Val Ser Ser Phe Gly435 440 445gcc agc ggt acg aac gcc cac atg atc ctg gaa cag gcg ccg gaa gaa 1392Ala Ser Gly Thr Asn Ala His Met Ile Leu Glu Gln Ala Pro Glu Glu450 455 460ccc gct gcg gag tcc ccc tcc gcc cca gcg ctc gat gga gtg gta ccg 1440Pro Ala Ala Glu Ser Pro Ser Ala Pro Ala Leu Asp Gly Val Val Pro465 470 475 480ctg gtg ctg tcg gcg gct acg gcc gcc tcg ctg acc ggc cag gcg gag 1488Leu Val Leu Ser Ala Ala Thr Ala Ala Ser Leu Thr Gly Gln Ala Glu485 490 495cga ctg ggg tcg ttc ctc gag gcg tcc ggc acg gtc gcg ctc gcc gat 1536Arg Leu Gly Ser Phe Leu Glu Ala Ser Gly Thr Val Ala Leu Ala Asp500 505 510gtg gcg gcc gca ctg gtc acc ggc cgg gcg tcg ctg gcc cag cgc gcg1584Val Ala Ala Ala Leu Val Thr Gly Arg Ala Ser Leu Ala Gln Arg Ala515 520 525gtc gtc gtg acc gac tcg ccc gag gag gcc ctg gca ggt ctc ggt gcg1632Val Val Val Thr Asp Ser Pro Glu Glu Ala Leu Ala Gly Leu Gly Ala530 535 540ctg gct cgt ggt gag gat gtt cgt ggg gtg gtt gct ggt ggt ggg gtg1680Leu Ala Arg Gly Glu Asp Val Arg Gly Val Val Ala Gly Gly Gly Val545 550 555 560agg tcg ggt ggg gac ggc aag gtt gtg ttg gtg ttt ccg ggt cag ggt1728Arg Ser Gly Gly Asp Gly Lys Val Val Leu Val Phe Pro Gly Gln Gly565 570 575tcg ccg tgg gtt ggt atg ggg cgt gag ttg ttg gag tgt tcg gag gtg1776Ser Pro Trp Val Gly Met Gly Arg Glu Leu Leu Glu Cys Ser Glu Val580 585 590ttt gcg gcg cgg gtg ggg gag tgt gcg gtg gcg ttg gag cgg tgg gtg1824Phe Ala Ala Arg Val Gly Glu Cys Ala Val Ala Leu Glu Arg Trp Val595 600 605gat tgg tcg ttg gtg gat gtg ttg cgg ggg gat tgt ccg gtt gag ttt1872Asp Trp Ser Leu Val Asp Val Leu Arg Gly Asp Cys Pro Val Glu Phe610 615 620ttt gag cgt gag gat gtg cgg cag ccg gcg agt ttt gcg gtg atg gtg1920Phe Glu Arg Glu Asp Val Arg Gln Pro Ala Ser Phe Ala Val Met Val625 630 635 640ggt ttg gcc gcg gtg tgg gag tcg gtg ggt gtg gtg gcg gat gcg gtg1968Gly Leu Ala Ala Val Trp Glu Ser Val Gly Val Val Ala Asp Ala Val645 650 655gtg ggt cat tcg ggt ggt gag gtt gct gct gcg tgt gtg tcg gkt gcg2016Val Gly His Ser Gly Gly Glu Val Ala Ala Ala Cys Val Ser Xaa Ala660 665 670ttg tcg ttg gag gac gct gtt cgg gtt gtg gcg gtc cgg agc aag acc2064Leu Ser Leu Glu Asp Ala Val Arg Val Val Ala Val Arg Ser Lys Thr675 680 685att tcc ggt gtt ctc tcg ggt cgg ggt ggt atg gcg tcg gtg ggg ttg2112Ile Ser Gly Val Leu Ser Gly Arg Gly Gly Met Ala Ser Val Gly Leu690 695 700tcg gag gag gag gcg gtt gct cgg cta cag cag tgg gat ggt cgg gtc2160Ser Glu Glu Glu Ala Val Ala Arg Leu Gln Gln Trp Asp Gly Arg Val705 710 715 720gag atc ggg gcg gtc aac agt ccg tcc tcg gtg gcc atc acc gct gac2208Glu Ile Gly Ala Val Asn Ser Pro Ser Ser Val Ala Ile Thr Ala Asp725 730 735acc gaa gcc ctc gac gaa gcc atc gag act ttg gag gac cag ggc gtc2256Thr Glu Ala Leu Asp Glu Ala Ile Glu Thr Leu Glu Asp Gln Gly Val740 745 750cgc gta cgc cgc atc gcg atc gac tac gcc tcg cac tcc cgg cac gtc2304Arg Val Arg Arg Ile Ala Ile Asp Tyr Ala Ser His Ser Arg His Val755 760 765ggg gct gtc cag gag atc ctg aac gag gca ttc gcc gac atc cgc agc2352Gly Ala Val Gln Glu Ile Leu Asn Glu Ala Phe Ala Asp Ile Arg Ser770 775 780caa gct cct acc gtg cca ttc ctc tcc acc gcc acc ggc gag tgg atc2400Gln Ala Pro Thr Val Pro Phe Leu Ser Thr Ala Thr Gly Glu Trp Ile785 790 795 800cgc gag gcg ggt gcc ctg gac ggc agc tac tgg tac cgc aac ctg cgc2448Arg Glu Ala Gly Ala Leu Asp Gly Ser Tyr Trp Tyr Arg Asn Leu Arg805 810 815agc cag gtc cgc ttc ggc ccc gcg atc gcc gac ctg ctg gcc gac ggc2496Ser Gln Val Arg Phe Gly Pro Ala Ile Ala Asp Leu Leu Ala Asp Gly820 825 830cac acc gtg ttc gtg gag tcc agc gcc cac ccc gtc ctg gtc cag ccg2544His Thr Val Phe Val Glu Ser Ser Ala His Pro Val Leu Val Gln Pro835 840 845atc agc gag gtc gtg gcc ggc gcc gag gca gag gcc gtc gtg acc ggc2592Ile Ser Glu Val Val Ala Gly Ala Glu Ala Glu Ala Val Val Thr Gly850 855 860tcc ctg cgc cgt cac gag gga ggt ccg cgc cgc ctg ttc act tcg atg2640Ser Leu Arg Arg His Glu Gly Gly Pro Arg Arg Leu Phe Thr Ser Met865 870 875 880gcc gac ctc ttc gtc cga ggc acc cac gtc gac tgg agc ggc gtc ctc2688Ala Asp Leu Phe Val Arg Gly Thr His Val Asp Trp Ser Gly Val Leu885 890 895gcg gcc gga gcc gat gcc cgc cgc gtc gac ctt ccg acg tac gcc ttt2736Ala Ala Gly Ala Asp Ala Arg Arg Val Asp Leu Pro Thr Tyr Ala Phe900 905 910gat cac aag aac tac tgg atg gag ctg gcc ggt acc gcc aac gat gtc2784Asp His Lys Asn Tyr Trp Met Glu Leu Ala Gly Thr Ala Asn Asp Val915 920 925gcc tcg ctc ggc ttg tcg ggg gct gat cat ccg ttg ctg ggt gcg gtg2832Ala Ser Leu Gly Leu Ser Gly Ala Asp His Pro Leu Leu Gly Ala Val930 935 940gtt ccg gtg ccg gag acg agc gga gtg ttg tgt acg tcg cgg ttg tcg2880Val Pro Val Pro Glu Thr Ser Gly Val Leu Cys Thr Ser Arg Leu Ser945 950 955 960ctt cgt aca cat ccg tgg ctt gcg gat cac gct gtg ggc cgg tgt cgt2928Leu Arg Thr His Pro Trp Leu Ala Asp His Ala Val Gly Arg Cys Arg965 970 975gct tgt ccc tgg cac tgc ttt ggt gga gtt ggt ggt gcg tgc ggg tga2976Ala Cys Pro Trp His Cys Phe Gly Gly Val Gly Gly Ala Cys Gly980 985 990<210>9<211>991<212>PRT<213>吸水鏈霉菌(Streptomyces hygroscopicus)<220><221>misc_feature<222>(41)<223>Xaa＝Ser.<220><221>misc_feature<222>(671)<223>Xaa＝Gly或Val.<400>9Met Thr Ala Pro Asp Glu Gln Ile Val Asp Ala Leu Arg Ala Ser Leu1 5 10 15Lys Glu Asn Met Arg Leu Gln Gln Glu Asn Gln Arg Leu Ser Glu Ser20 25 30Ser Ala Glu Pro Ile Ala Ile Val Xaa Met Ala Cys Arg Tyr Ala Gly35 40 45Gly Ile Arg Asn Pro Glu Asp Leu Trp Arg Val Val Asn Asp Gly Thr50 55 60Asp Val Tyr Thr Ser Phe Pro Glu Asn Arg Gly Trp Asp Leu Glu Gly65 70 75 80Ile Tyr His Pro Asp Pro Asp Asn Pro Gly Thr Thr Tyr Val Arg Glu85 90 95Gly Ala Phe Leu His Asp Ala Asn Leu Phe Asp Ala Gly Leu Phe Gly100 105 110Ile Ser Pro Leu Glu Ala Leu Ala Met Glu Pro Gln Gln Arg Gln Leu115 120 125Leu Glu Ile Cys Trp Glu Ala Leu Glu Arg Ala Gly Ile Asp Pro His130 135 140Ser Val Arg Gly Ala Asp Ile Gly Val Tyr Ala Gly Leu Val His Gln145 150 155 160Asp Tyr Ala Pro Asp Leu Ser Gly Leu Glu Gly Tyr Leu Ser Leu Glu165 170 175Arg Ala Leu Gly Ser Ala Gly Gly Ile Ala Ser Gly Arg Val Ala Tyr180 185 190Thr Leu Gly Leu Glu Gly Pro Ala Val Thr Val Asp Thr Met Cys Ser195 200 205Ser Thr Leu Val Ala Val His Val Ala Thr Gln Ala Leu Arg Arg Gly210 215 220Glu Cys Ala Met Ala Leu Ala Gly Gly Ala Thr Val Met Ser Thr Pro225 230 235 240Gly Gly Phe Ile Gly Phe Ala Arg Gln Arg Ala Leu Ala Phe Asp Gly245 250 255Arg Cys Lys Ser Tyr Gly Ala Ala Ala Asp Gly Ser Ser Trp Ala Glu260 265 270Gly Ala Gly Val Val Leu Leu Glu Arg Leu Ser Asp Ala Arg Arg Asn275 280 285Gly His Arg Val Leu Ala Val Ile Arg Gly Ser Ala Leu Asn Gln Asp290 295 300Gly Ala Ser Asn Gly Leu Thr Ala Pro Asn Gly Pro Ala Gln Arg Arg305 310 315 320Val Ile Arg Lys Ala Leu Glu Asn Ala Gly Leu Thr Thr Ala Asp Ile325 330 335Asp Met Val Glu Gly His Gly Thr Gly Thr Val Leu Gly Asp Pro Ile340 345350Glu Ala Gln Ala Leu Ile Ala Thr Tyr Gly Gln Asp Arg Pro Glu Gly355360 365Arg Pro Leu Trp Leu Gly Ser Val Lys Ser Val Ile Gly His Thr Gln370 375 380Ser Gly Ser Gly Val Ala Gly Leu Ile Asn Ala Val Gln Ala Leu Arg385 390 395 400His Gly Val Met Pro Ala Thr Arg His Val Asp Ala Pro Asn Pro Gln405 410 415Val Asp Trp Ser Ala Gly Ala Val Glu Leu Leu Thr Glu Ala Arg Ala420 425 430Trp Pro Glu Leu Gly Arg Pro Arg Arg Ala Gly Val Ser Ser Phe Gly435 440 445Ala Ser Gly Thr Asn Ala His Met Ile Leu Glu Gln Ala Pro Glu Glu450 455 460Pro Ala Ala Glu Ser Pro Ser Ala Pro Ala Leu Asp Gly Val Val Pro465 470 475 480Leu Val Leu Ser Ala Ala Thr Ala Ala Ser Leu Thr Gly Gln Ala Glu485 490 495Arg Leu Gly Ser Phe Leu Glu Ala Ser Gly Thr Val Ala Leu Ala Asp500 505 510Val Ala Ala Ala Leu Val Thr Gly Arg Ala Ser Leu Ala Gln Arg Ala515 520 525Val Val Val Thr Asp Ser Pro Glu Glu Ala Leu Ala Gly Leu Gly Ala530 535 540Leu Ala Arg Gly Glu Asp Val Arg Gly Val Val Ala Gly Gly Gly Val545 550 555 560Arg Ser Gly Gly Asp Gly Lys Val Val Leu Val Phe Pro Gly Gln Gly565 570 575Ser Pro Trp Val Gly Met Gly Arg Glu Leu Leu Glu Cys Ser Glu Val580 585 590Phe Ala Ala Arg Val Gly Glu Cys Ala Val Ala Leu Glu Arg Trp Val595 600 605Asp Trp Ser Leu Val Asp Val Leu Arg Gly Asp Cys Pro Val Glu Phe610 615 620Phe Glu Arg Glu Asp Val Arg Gln Pro Ala Ser Phe Ala Val Met Val625 630 635 640Gly Leu Ala Ala Val Trp Glu Ser Val Gly Val Val Ala Asp Ala Val645 650 655Val Gly His Ser Gly Gly Glu Val Ala Ala Ala Cys Val Ser Xaa Ala660 665 670Leu Ser Leu Glu Asp Ala Val Arg Val Val Ala Val Arg Ser Lys Thr675 680 685Ile Ser Gly Val Leu Ser Gly Arg Gly Gly Met Ala Ser Val Gly Leu
690 695 700Ser Glu Glu Glu Ala Val Ala Arg Leu Gln Gln Trp Asp Gly Arg Val705 710 715 720Glu Ile Gly Ala Val Ash Ser Pro Ser Ser Val Ala Ile Thr Ala Asp725 730 735Thr Glu Ala Leu Asp Glu Ala Ile Glu Thr Leu Glu Asp Gln Gly Val740 745 750Arg Val Arg Arg Ile Ala Ile Asp Tyr Ala Ser His Ser Arg His Val755 760765Gly Ala Val Gln Glu Ile Leu Asn Glu Ala Phe Ala Asp Ile Arg Ser770 775 780Gln Ala Pro Thr Val Pro Phe Leu Ser Thr Ala Thr Gly Glu Trp Ile785 790 795 800Arg Glu Ala Gly Ala Leu Asp Gly Ser Tyr Trp Tyr Arg Asn Leu Arg805 810 815Ser Gln Val Arg Phe Gly Pro Ala Ile Ala Asp Leu Leu Ala Asp Gly820 825 830His Thr Val Phe Val Glu Ser Ser Ala His Pro Val Leu Val Gln Pro835 840 845Ile Ser Glu Val Val Ala Gly Ala Glu Ala Glu Ala Val Val Thr Gly850 855 860Ser Leu Arg Arg His Glu Gly Gly Pro Arg Arg Leu Phe Thr Ser Met865 870 875 880Ala Asp Leu Phe Val Arg Gly Thr His Val Asp Trp Ser Gly Val Leu885 890 895Ala Ala Gly Ala Asp Ala Arg Arg Val Asp Leu Pro Thr Tyr Ala Phe900 905 910Asp His Lys Asn Tyr Trp Met Glu Leu Ala Gly Thr Ala Asn Asp Val915 920 925Ala Ser Leu Gly Leu Ser Gly Ala Asp His Pro Leu Leu Gly Ala Val930 935 940Val Pro Val Pro Glu Thr Ser Gly Val Leu Cys Thr Ser Arg Leu Ser945 950 955 960Leu Arg Thr His Pro Trp Leu Ala Asp His Ala Val Gly Arg Cys Arg965 970 975Ala Cys Pro Trp His Cys Phe Gly Gly Val Gly Gly Ala Cys Gly980 985 990<210>10<211>1407<212>DNA<213>吸水鏈霉菌(Streptomyces hygroscopicus)<220><221>CDS<222>(1)..(1407)<223>shnE<400>10atg ccc gcc gcg tcg acc ttc cga cgt acg cct ttg atc aca aga act 48Met Pro Ala Ala Ser Thr Phe Arg Arg Thr Pro Leu Ile Thr Arg Thr1 5 10 15act gga tgg agc tgg ccg gta ccg cca acg atg tcg cct cgc tcg gct 96Thr Gly Trp Ser Trp Pro Val Pro Pro Thr Met Ser Pro Arg Ser Ala20 25 30tgt cgg ggg ctg atc atc cgt tgc tgg gtg cgg tgg ttc cgg tgc cgg144Cys Arg Gly Leu Ile Ile Arg Cys Trp Val Arg Trp Phe Arg Cys Arg35 40 45aga cga gcg gag tgt tgt gta cgt cgc ggt tgt cgc ttc gta cac atc192Arg Arg Ala Glu Cys Cys Val Arg Arg Gly Cys Arg Phe Val His Ile50 55 60cgt ggc ttg cgg atc acg ctg tgg gcc ggt gtc gtg ctt gtc cct ggc240Arg Gly Leu Arg Ile Thr Leu Trp Ala Gly Val Val Leu Val Pro Gly65 70 75 80act gct ttg gtg gag ttg gtg gtg cgt gcg ggt gac aag gtg ggc tgc288Thr Ala Leu Val Glu Leu Val Val Arg Ala Gly Asp Lys Val Gly Cys85 90 95ggc acg ttg gag gaa ttg gtc atc gag acg ccg ctt gtc gta ccc gcg336Gly Thr Leu Glu Glu Leu Val Ile Glu Thr Pro Leu Val Val Pro Ala100 105 110caa ggg agt atg cgc gtt cag ttc gcg gtg ggc ggc cct gag gag aac384Gln Gly Ser Met Arg Val Gln Phe Ala Val Gly Gly Pro Glu Glu Asn115 120 125ggc gcg cgt tcg gtg gcc gtg tac tcg gct cgt gat gac gac ggt cgc432Gly Ala Arg Ser Val Ala Val Tyr Ser Ala Arg Asp Asp Asp Gly Arg130 135 140ggc acc ggt atc gat ggt tgg acc cgt cac gcc gcc ggc act ctg acg480Gly Thr Gly Ile Asp Gly Trp Thr Arg His Ala Ala Gly Thr Leu Thr145 150 155 160gcg gct gct gtc cct gct gat ggt ttc gat ttc acg gtg tgg ccg ccg528Ala Ala Ala Val Pro Ala Asp Gly Phe Asp Phe Thr Val Trp Pro Pro165 170 175gtc ggt gcg gag cgg gtg tcg ttc gat gcg gtc ggg ttc tat gag gag576Val Gly Ala Glu Arg Val Ser Phe Asp Ala Val Gly Phe Tyr Glu Glu180 185 190atg gcg ggc cgc ggc tat gtg tac ggt ccg gcg ttc cag ggt ttg cgt624Met Ala Gly Arg Gly Tyr Val Tyr Gly Pro Ala Phe Gln Gly Leu Arg195 200 205ggg gtg tgg cgg cgg ggc gaa gag gtg ttc gcc gag gtc gct ctg ccg672Gly Val Trp Arg Arg Gly Glu Glu Val Phe Ala Glu Val Ala Leu Pro210 215 220gac gag cag cat ggt gag gcg agc cgc ttc ggg ttg cac ccg gcg ttg720Asp Glu Gln His Gly Glu Ala Ser Arg Phe Gly Leu His Pro Ala Leu225 230 235 240ctc gac gcc gcc ttg cag agc ggg ctc gtc cgg ccg gcc gat gcc ggg768Leu Asp Ala Ala Leu Gln Ser Gly Leu Val Arg Pro Ala Asp Ala Gly245 250 255gtg gat atg cgt gtg ccg ttc gcc tgg aac ggg ctg cgc ctg cat gcc816Val Asp Met Arg Val Pro Phe Ala Trp Asn Gly Leu Arg Leu His Ala
260 265 270gcg ggt gcc tcg gag ttg cgg gtg cgg acg gtg ccg tcc ggg ccg gac 864Ala Gly Ala Ser Glu Leu Arg Val Arg Thr Val Pro Ser Gly Pro Asp275 280 285gcg gtg tcg ttg cag gcg gcc gac ggg gcc ggc ggt ccg gtg ctg agc 912Ala Val Ser Leu Gln Ala Ala Asp Gly Ala Gly Gly Pro Val Leu Ser290 295 300ctg gag tcg ctg gtt gcc cgg gcg gtg gac gtg gag caa ctg gat cgg 960Leu Glu Ser Leu Val Ala Arg Ala Val Asp Val Glu Gln Leu Asp Arg305 310 315 320atg gcg act gat gac ggt cgc gac gcg ctg ttc gag gtg gac tgg agc1008Met Ala Thr Asp Asp Gly Arg Asp Ala Leu Phe Glu Val Asp Trp Ser325 330 335gaa ctg ccc gcg cct gct tcg agc gtg gag tct ctg ccg ccg tcg gcg1056Glu Leu Pro Ala Pro Ala Ser Ser Val Glu Ser Leu Pro Pro Ser Ala340 345 350ctg gtg gct tcg gcc gag gac gtg acg gat ctg gca gat gcc gcg gtg1104Leu Val Ala Ser Ala Glu Asp Val Thr Asp Leu Ala Asp Ala Ala Val355 360 365gtt cct gcg gta gcg gtt ctt gag gct gtt ggc ggt gac ggc gag cac1152Val Pro Ala Val Ala Val Leu Glu Ala Val Gly Gly Asp Gly Glu His370 375 380gac gcg ctt gcc ctg acc gtc agg gtg ctg gag gtc gtc cag gcg tgg1200Asp Ala Leu Ala Leu Thr Val Arg Val Leu Glu Val Val Gln Ala Trp385 390 395 400ttc gct gct gcg ggt ctg gcg gag tcc cgg ctg gtg gtg gtc aca cgg1248Phe Ala Ala Ala Gly Leu Ala Glu Ser Arg Leu Val Val Val Thr Arg405 410 415ggt gcg gtt ccg gtc ggc ggt gag gga aat gtc gcc gat ccc gct ggt1296Gly Ala Val Pro Val Gly Gly Glu Gly Asn Val Ala Asp Pro Ala Gly420 425 430gct gcg gtg tgg ggt ctg gtt cgg gcg gcc cag gcc gag aac ccg gac1344Ala Ala Val Trp Gly Leu Val Arg Ala Ala Gln Ala Glu Asn Pro Asp435440 445cgg atc gtc ctg ctc gat ctc gct gcc gac gtc gat atg gga tcg gtt1392Arg Ile Val Leu Leu Asp Leu Ala Ala Asp Val Asp Met Gly Ser Val450 455 460ctg cct gcc gta ttg1407Leu Pro Ala Val Leu465<210>11<211>469<212>PRT<213>吸水鏈霉菌(Streptomyces hygroscopicus)<400>11Met Pro Ala Ala Ser Thr Phe Arg Arg Thr Pro Leu Ile Thr Arg Thr1 5 10 15Thr Gly Trp Ser Trp Pro Val Pro Pro Thr Met Ser Pro Arg Ser Ala
20 25 30Cys Arg Gly Leu Ile Ile Arg Cys Trp Val Arg Trp Phe Arg Cys Arg35 40 45Arg Arg Ala Glu Cys Cys Val Arg Arg Gly Cys Arg Phe Val His Ile50 55 60Arg Gly Leu Arg Ile Thr Leu Trp Ala Gly Val Val Leu Val Pro Gly65 70 75 80Thr Ala Leu Val Glu Leu Val Val Arg Ala Gly Asp Lys Val Gly Cys85 90 95Gly Thr Leu Glu Glu Leu Val Ile Glu Thr Pro Leu Val Val Pro Ala100 105 110Gln Gly Ser Met Arg Val Gln Phe Ala Val Gly Gly Pro Glu Glu Asn115 120 125Gly Ala Arg Ser Val Ala Val Tyr Ser Ala Arg Asp Asp Asp Gly Arg130 135 140Gly Thr Gly Ile Asp Gly Trp Thr Arg His Ala Ala Gly Thr Leu Thr145 150 155 160Ala Ala Ala Val Pro Ala Asp Gly Phe Asp Phe Thr Val Trp Pro Pro165 170 175Val Gly Ala Glu Arg Val Ser Phe Asp Ala Val Gly Phe Tyr Glu Glu180 185 190Met Ala Gly Arg Gly Tyr Val Tyr Gly Pro Ala Phe Gln Gly Leu Arg195 200 205Gly Val Trp Arg Arg Gly Glu Glu Val Phe Ala Glu Val Ala Leu Pro210 215 220Asp Glu Gln His Gly Glu Ala Ser Arg Phe Gly Leu His Pro Ala Leu225 230 235 240Leu Asp Ala Ala Leu Gln Ser Gly Leu Val Arg Pro Ala Asp Ala Gly245 250 255Val Asp Met Arg Val Pro Phe Ala Trp Asn Gly Leu Arg Leu His Ala260 265 270Ala Gly Ala Ser Glu Leu Arg Val Arg Thr Val Pro Ser Gly Pro Asp275 280 285Ala Val Ser Leu Gln Ala Ala Asp Gly Ala Gly Gly Pro Val Leu Ser290 295 300Leu Glu Ser Leu Val Ala Arg Ala Val Asp Val Glu Gln Leu Asp Arg305 310 315 320Met Ala Thr Asp Asp Gly Arg Asp Ala Leu Phe Glu Val Asp Trp Ser325 330 335Glu Leu Pro Ala Pro Ala Ser Ser Val Glu Ser Leu Pro Pro Ser Ala340 345 350Leu Val Ala Ser Ala Glu Asp Val Thr Asp Leu Ala Asp Ala Ala Val355 360 365Val Pro Ala Val Ala Val Leu Glu Ala Val Gly Gly Asp Gly Glu His370 375 380Asp Ala Leu Ala Leu Thr Val Arg Val Leu Glu Val Val Gln Ala Trp385 390 395 400Phe Ala Ala Ala Gly Leu Ala Glu Ser Arg Leu Val Val Val Thr Arg405 410 415Gly Ala Val Pro Val Gly Gly Glu Gly Asn Val Ala Asp Pro Ala Gly420 425 430Ala Ala Val Trp Gly Leu Val Arg Ala Ala Gln Ala Glu Asn Pro Asp435 440 445Arg Ile Val Leu Leu Asp Leu Ala Ala Asp Val Asp Met Gly Ser Val450 455 460Leu Pro Ala Val Leu465<210>12<211>834<212>DNA<213>吸水鏈霉菌(Streptomyces hygroscopicus)<220><221>CDS<222>(1)..(834)<223>shnN<400>12gtg cgg agc tgg acc aga gcc aat gta aac gtt gtt tcc ttc tcg gat48Val Arg Ser Trp Thr Arg Ala Asn Val Asn Val Val Ser Phe Ser Asp1 5 10 15gaa gtg gag ttc ggc gtg ttc gtc aaa agg gac tat ctt cag cgc ata96Glu Val Glu Phe Gly Val Phe Val Lys Arg Asp Tyr Leu Gln Arg Ile20 25 30ggt tac gaa gga tcc ggt att ccg aac ctt cag acc ctg gcg gaa ctt 144Gly Tyr Glu Gly Ser Gly Ile Pro Asn Leu Gln Thr Leu Ala Glu Leu35 40 45cag tgg ctg cat ctc tgt agc ctc ccc tac gac acc ggt tac att ctg 192Gln Trp Leu His Leu Cys Ser Leu Pro Tyr Asp Thr Gly Tyr Ile Leu50 55 60cat cag ccg tac gag gat ttt gac atg ccc cgc gta ttc gaa gcg gtg 240His Gln Pro Tyr Glu Asp Phe Asp Met Pro Arg Val Phe Glu Ala Val65 70 75 80atg aaa cgg ggc gga gtc tgt ttc gag ttg aat ttc ctc ttc cat cgt 288Met Lys Arg Gly Gly Val Cys Phe Glu Leu Asn Phe Leu Phe His Arg85 90 95ctc ctt gtc gag atg gga ttc gat gcg cat gtg aat tcc gcc agc acg 336Leu Leu Val Glu Met Gly Phe Asp Ala His Val Asn Ser Ala Ser Thr100 105 110gct ctc ccc ggc ggc cag tgg ggt tcc gag atc gag cae atg gct atc 384Ala Leu Pro Gly Gly Gln Trp Gly Ser Glu Ile Glu His Met Ala Ile115 120 125cgt gtc cgt ata gac gac gtg gat tgg ctc gtc gac gtc ggg cac gga 432Arg Val Arg Ile Asp Asp Val Asp Trp Leu Val Asp Val Gly His Gly130 135 140agc gtg gcc atc acg gag ccc ctg cgt atc gat gaa cag gcg gga agc 480Ser Val Ala Ile Thr Glu Pro Leu Arg Ile Asp Glu Gln Ala Gly Ser145 150 155 160gtg gtt cag atg ggc acg gag ttc cgc ttg gcc acg cgg ggc gag tgg528Val Val Gln Met Gly Thr Glu Phe Arg Leu Ala Thr Arg Gly Glu Trp165 170 175cgc gtc ctt caa tac aag cca aag ggc agg gat tgg cgt gac gca tat576Arg Val Leu Gln Tyr Lys Pro Lys Gly Arg Asp Trp Arg Asp Ala Tyr180 185 190cgg atg aaa atc aaa gat cgt gcc att tcc gat tgg aat aca tgg cga624Arg Met Lys Ile Lys Asp Arg Ala Ile Ser Asp Trp Asn Thr Trp Arg195 200 205gaa gaa ctg ccg ccc gac gcg gac cct gtg gtg ccg cgg aag cgg cga672Glu Glu Leu Pro Pro Asp Ala Asp Pro Val Val Pro Arg Lys Arg Arg210 215 220cgc ggt gtg gag aac ggg cag gtg acc ctc gtc gcc aat ctc ttc agg720Arg Gly Val Glu Asn Gly Gln Val Thr Leu Val Ala Asn Leu Phe Arg225 230 235 240tcc atc atc ggg ggc gag gag acg gtg aag cac gta cgt gat gaa gca768Ser Ile Ile Gly Gly Glu Glu Thr Val Lys His Val Arg Asp Glu Ala245 250 255gag ctg atc gag atc atg act act tac tgg gga gag tcc gca cct atc816Glu Leu Ile Glu Ile Met Thr Thr Tyr Trp Gly Glu Ser Ala Pro Ile260 265 270gtc ggg tac gaa cga tga834Val Gly Tyr Glu Arg275<210>13<211>277<212>PRT<213>吸水鏈霉菌(Streptomyces hygroscopicus)<400>13Val Arg Ser Trp Thr Arg Ala Asn Val Asn Val Val Ser Phe Ser Asp1 5 10 15Glu Val Glu Phe Gly Val Phe Val Lys Arg Asp Tyr Leu Gln Arg Ile20 25 30Gly Tyr Glu Gly Ser Gly Ile Pro Asn Leu Gln Thr Leu Ala Glu Leu35 40 45Gln Trp Leu His Leu Cys Ser Leu Pro Tyr Asp Thr Gly Tyr Ile Leu50 55 60His Gln Pro Tyr Glu Asp Phe Asp Met Pro Arg Val Phe Glu Ala Val65 70 75 80Met Lys Arg Gly Gly Val Cys Phe Glu Leu Asn Phe Leu Phe His Arg85 90 95Leu Leu Val Glu Met Gly Phe Asp Ala His Val Asn Ser Ala Ser Thr100 105 110Ala Leu Pro Gly Gly Gln Trp Gly Ser Glu Ile Glu His Met Ala Ile115 120 125Arg Val Arg Ile Asp Asp Val Asp Trp Leu Val Asp Val Gly His Gly
130 135 140Ser Val Ala Ile Thr Glu Pro Leu Arg Ile Asp Glu Gln Ala Gly Ser145 150 155 160Val Val Gln Met Gly Thr Glu Phe Arg Leu Ala Thr Arg Gly Glu Trp165 170 175Arg Val Leu Gln Tyr Lys Pro Lys Gly Arg Asp Trp Arg Asp Ala Tyr180 185 190Arg Met Lys Ile Lys Asp Arg Ala Ile Ser Asp Trp Asn Thr Trp Arg195 200 205Glu Glu Leu Pro Pro Asp Ala Asp Pro Val Val Pro Arg Lys Arg Arg210 215 220Arg Gly Val Glu Asn Gly Gln Val Thr Leu Val Ala Asn Leu Phe Arg225 230 235 240Ser Ile Ile Gly Gly Glu Glu Thr Val Lys His Val Arg Asp Glu Ala245 250 255Glu Leu Ile Glu Ile Met Thr Thr Tyr Trp Gly Glu Ser Ala Pro Ile260 265 270Val Gly Tyr Glu Arg275<210>14<211>1101<212>DNA<213>吸水鏈霉菌(Streptomyces hygroscopicus)<220><221>CDS<222>(1)..(1101)<223>shnO<400>14gtg agg gtc cgg gct gcc gtc gtc ggg ctc ggc tgg gcg ggc cgc gac 48Val Arg Val Arg Ala Ala Val Val Gly Leu Gly Trp Ala Gly Arg Asp1 5 10 15ctg tgg ctc aga ctg ctg cgc gag cac aag gac ttc gag gtg gtc gcc 96Leu Trp Leu Arg Leu Leu Arg Glu His Lys Asp Phe Glu Val Val Ala20 25 30ggt gtc gac ccc gac ccg gac tcc cgg gcg gcc get gtg gcc acc ggt144Gly Val Asp Pro Asp Pro Asp Ser Arg Ala Ala Ala Val Ala Thr Gly35 40 45ctg cgc gcc cac ccc acc gtg gac gcc ctc gat ccc cgc acg gtc gac192Leu Arg Ala His Pro Thr Val Asp Ala Leu Asp Pro Arg Thr Val Asp50 55 60atc gcc gtc gtc gcg gta ccc aac cat ctg cat gcc gag gtt gcg gcc240Ile Ala Val Val Ala Val Pro Asn His Leu His Ala Glu Val Ala Ala65 70 75 80gcc ctg ctc cgc cga ggg atc tcc gtc ttt ctg gag aag ccg gtc tgc288Ala Leu Leu Arg Arg Gly Ile Ser Val Phe Leu Glu Lys Pro Val Cys85 90 95ctc acc act gcc gag gcc gac gcc ctc gcc gag gcc gaa ggc aac ggc336Leu Thr Thr Ala Glu Ala Asp Ala Leu Ala Glu Ala Glu Gly Asn Gly
l00 105 110gcg gtg ctg ctc gcc ggc agt gcc gcc gcc cac cgc ggc gac atc cgc384Ala Val Leu Leu Ala Gly Ser Ala Ala Ala His Arg Gly Asp Ile Arg115 120 125gag ctc tcc ggg ctg ctt ccc cag ctc ggc cgg atc cgc cat gcc gac432Glu Leu Ser Gly Leu Leu Pro Gln Leu Gly Arg Ile Arg His Ala Asp130 135 140ctg tcc tgg gtc agg gcg cgc gga gtg ccg cag ccc ggc ggc tgg ttc480Leu Ser Trp Val Arg Ala Arg Gly Val Pro Gln Pro Gly Gly Trp Phe145 150 155 160acc cag cgc agc agg gcc ggt ggc ggt gcg ctc gtc gac ctc ggc tgg528Thr Gln Arg Ser Arg Ala Gly Gly Gly Ala Leu Val Asp Leu Gly Trp165 170 175cac ctt ctc gac gtc ctc gcc ttc ctc ctc ggc ccc gcc ccc gtc gcc576His Leu Leu Asp Val Leu Ala Phe Leu Leu Gly Pro Ala Pro Val Ala180 185 190cag gtg atc ggc tcg atc tcc gac gac ttc gtc agc agc aga gcc tgg624Gln Val Ile Gly Ser Ile Ser Asp Asp Phe Val Ser Ser Arg Ala Trp195 200 205tcc gcc acg tgg cgt gag gac cag ctc acc gac gcc ccg acc ggc gac672Ser Ala Thr Trp Arg Glu Asp Gln Leu Thr Asp Ala Pro Thr Gly Asp210 215 220gtc gag gac acc gcc cgc ggc ttc ctg gtc cgc gag gac ggt atc tct720Val Glu Asp Thr Ala Arg Gly Phe Leu Val Arg Glu Asp Gly Ile Ser225 230 235 240gtc tcg ttg cgg gcc agt tgg gcc tca cac gag gcg ctg gac ggc tcc768Val Ser Leu Arg Ala Ser Trp Ala Ser His Glu Ala Leu Asp Gly Ser245 250 255gtc atc acc atc gag ggc agc gat gga acg gca cgg ctg cac tgc acc816Val Ile Thr Ile Glu Gly Ser Asp Gly Thr Ala Arg Leu His Cys Thr260 265 270ttc ggc ttc agc ccg aac cgg gct ccc gaa tcg gtg ctc acc ctc acg864Phe Gly Phe Ser Pro Asn Arg Ala Pro Glu Ser Val Leu Thr Leu Thr275 280 285cag gat ggc tcc aca cag cgg atc ccg ctg ccc gcc gaa ccc atc ggc912Gln Asp Gly Ser Thr Gln Arg Ile Pro Leu Pro Ala Glu Pro Ile Gly290 295 300att gag tac ggt cgc cag ctc gac ggc ctc gcc cgg ctc ctg gcc gac960Ile Glu Tyr Gly Arg Gln Leu Asp Gly Leu Ala Arg Leu Leu Ala Asp305 310 315 320ccg ggc cga cgg ggc cag gcc gtc gcc cag gcc cgc agc acc gtc cgg 1008Pro Gly Arg Arg Gly Gln Ala Val Ala Gln Ala Arg Ser Thr Val Arg325 330 335ttg atc gag agc ttc tat gca tcg gcg cgc gct gcc cca cct gtg gat 1056Leu Ile Glu Ser Phe Tyr Ala Ser Ala Arg Ala Ala Pro Pro Val Asp340 345 350cac gcg tcc gaa ttc acc gcc cac aaa gag gtg agg atc gca tga 1101His Ala Ser Glu Phe Thr Ala His Lys Glu Val Arg Ile Ala
355 360 365<210>15<211>366<212>PRT<213>吸水鏈霉菌(Streptomyces hygroscopicus)<400>15Val Arg Val Arg Ala Ala Val Val Gly Leu Gly Trp Ala Gly Arg Asp1 5 10 15Leu Trp Leu Arg Leu Leu Arg Glu His Lys Asp Phe Glu Val Val Ala20 25 30Gly Val Asp Pro Asp Pro Asp Ser Arg Ala Ala Ala Val Ala Thr Gly35 40 45Leu Arg Ala His Pro Thr Val Asp Ala Leu Asp Pro Arg Thr Val Asp50 55 60Ile Ala Val Val Ala Val Pro Asn His Leu His Ala Glu Val Ala Ala65 70 75 80Ala Leu Leu Arg Arg Gly Ile Ser Val Phe Leu Glu Lys Pro Val Cys85 90 95Leu Thr Thr Ala Glu Ala Asp Ala Leu Ala Glu Ala Glu Gly Asn Gly100 105 110Ala Val Leu Leu Ala Gly Ser Ala Ala Ala His Arg Gly Asp Ile Arg115 120 125Glu Leu Ser Gly Leu Leu Pro Gln Leu Gly Arg Ile Arg His Ala Asp130 135 140Leu Ser Trp Val Arg Ala Arg Gly Val Pro Gln Pro Gly Gly Trp Phe145 150 155 160Thr Gln Arg Ser Arg Ala Gly Gly Gly Ala Leu Val Asp Leu Gly Trp165 170 175His Leu Leu Asp Val Leu Ala Phe Leu Leu Gly Pro Ala Pro Val Ala180 185 190Gln Val Ile Gly Ser Ile Ser Asp Asp Phe Val Ser Ser Arg Ala Trp195 200 205Ser Ala Thr Trp Arg Glu Asp Gln Leu Thr Asp Ala Pro Thr Gly Asp210 215 220Val Glu Asp Thr Ala Arg Gly Phe Leu Val Arg Glu Asp Gly Ile Ser225 230 235 240Val Ser Leu Arg Ala Ser Trp Ala Ser His Glu Ala Leu Asp Gly Ser245 250 255Val Ile Thr Ile Glu Gly Ser Asp Gly Thr Ala Arg Leu His Cys Thr260 265 270Phe Gly Phe Ser Pro Asn Arg Ala Pro Glu Ser Val Leu Thr Leu Thr275 280 285Gln Asp Gly Ser Thr Gln Arg Ile Pro Leu Pro Ala Glu Pro Ile Gly290 295 300Ile Glu Tyr Gly Arg Gln Leu Asp Gly Leu Ala Arg Leu Leu Ala Asp305 310 315 320Pro Gly Arg Arg Gly Gln Ala Val Ala Gln Ala Arg Ser Thr Val Arg325 330 335Leu Ile Glu Ser Phe Tyr Ala Ser Ala Arg Ala Ala Pro Pro Val Asp340 345 350His Ala Ser Glu Phe Thr Ala His Lys Glu Val Arg Ile Ala355 360 365<210>16<211>699<212>DNA<213>吸水鏈霉菌(Streptomyces hygroscopicus)<220><221>CDS<222>(1)..(699)<223>shnP<400>16atg acc aga ggg ccg gaa ttc cat gcg gca ccg acc atc gaa cag cgc 48Met Thr Arg Gly Pro Glu Phe His Ala Ala Pro Thr Ile Glu Gln Arg1 5 10 15ccc ccg gtc ccc aga cac gct gtc atc ttc gat ctc gac ggg gtc gtc 96Pro Pro Val Pro Arg His Ala Val Ile Phe Asp Leu Asp Gly Val Val20 25 30gtc gac agc ttc gag gtg atg ggt gag gcg ttc tcc ctg gcg tac gcc144Val Asp Ser Phe Glu Val Met Gly Glu Ala Phe Ser Leu Ala Tyr Ala35 40 45gag gtc gtc ggc acc ggc gag gca cct ttc gag gag tac cgg cgc cac192Glu Val Val Gly Thr Gly Glu Ala Pro Phe Glu Glu Tyr Arg Arg His50 55 60cag ggc cgc tac ttt ccc gac atc atg cgg atc atg ggc ctt ccg ctg240Gln Gly Arg Tyr Phe Pro Asp Ile Met Arg Ile Met Gly Leu Pro Leu65 70 75 80gag atg gaa gag ccc ttc gtc cgc gag agc tac cgg ctc gcc gac cgc288Glu Met Glu Glu Pro Phe Val Arg Glu Ser Tyr Arg Leu Ala Asp Arg85 90 95gtc cag gtg tac gac ggt gtc gtc gac gtc ctg cgg acg ctg aac gaa336Val Gln Val Tyr Asp Gly Val Val Asp Val Leu Arg Thr Leu Asn Glu100 105 110cgc ggc ctc cgg ctg gcc atc gcc acc ggc aag gca ggc gag cgc gcc384Arg Gly Leu Arg Leu Ala Ile Ala Thr Gly Lys Ala Gly Glu Arg Ala115 120 125cgg tcc ctg ctc gat gtc ctc ggc ctg ctc ccg tac ttc gcc cac gtc432Arg Ser Leu Leu Asp Val Leu Gly Leu Leu Pro Tyr Phe Ala His Val130 135 140atc ggc tcc gac gag gtg ccc cgg ccc aag ccc gcc cct gac atc atc480Ile Gly Ser Asp Glu Val Pro Arg Pro Lys Pro Ala Pro Asp Ile Ile145 150 155 160aga cgc gca ctc gaa ctc ctc gag gtt ccg gcg gag cgg gcc atc atg528Arg Arg Ala Leu Glu Leu Leu Glu Val Pro Ala Glu Arg Ala Ile Met165 170 175atc ggc gac gcc ccc acc gac ctg gcc agc gcc cac ggc gcc gac gtc576Ile Gly Asp Ala Pro Thr Asp Leu Ala Ser Ala His Gly Ala Asp Val
180 185 190acc gcc gta gcc gcg ctg tgg ggc tgc cag gaa ggg gcc gaa ctg ctc624Thr Ala Val Ala Ala Leu Trp Gly Cys Gln Glu Gly Ala Glu Leu Leu195 200 205gcc gcc gac ccc gat gtc gtc ctg cgg tgg ccc gcc gac ctg ctc gcc672Ala Ala Asp Pro Asp Val Val Leu Arg Trp Pro Ala Asp Leu Leu Ala210 215 220ctc tgc ccg gcc ctg ccc ggc cac tga699Leu Cys Pro Ala Leu Pro Gly His225 230<210>17<211>232<212>PRT<213>吸水鏈霉菌(Streptomyces hygroscopicus)<400>17Met Thr Arg Gly Pro Glu Phe His Ala Ala Pro Thr Ile Glu Gln Arg1 5 10 15Pro Pro Val Pro Arg His Ala Val Ile Phe Asp Leu Asp Gly Val Val20 25 30Val Asp Ser Phe Glu Val Met Gly Glu Ala Phe Ser Leu Ala Tyr Ala35 40 45Glu Val Val Gly Thr Gly Glu Ala Pro Phe Glu Glu Tyr Arg Arg His50 55 60Gln Gly Arg Tyr Phe Pro Asp Ile Met Arg Ile Met Gly Leu Pro Leu65 70 75 80Glu Met Glu Glu Pro Phe Val Arg Glu Ser Tyr Arg Leu Ala Asp Arg85 90 95Val Gln Val Tyr Asp Gly Val Val Asp Val Leu Arg Thr Leu Asn Glu100 105 110Arg Gly Leu Arg Leu Ala Ile Ala Thr Gly Lys Ala Gly Glu Arg Ala115 120 125Arg Ser Leu Leu Asp Val Leu Gly Leu Leu Pro Tyr Phe Ala His Val130 135 140Ile Gly Ser Asp Glu Val Pro Arg Pro Lys Pro Ala Pro Asp Ile Ile145 150 155 160Arg Arg Ala Leu Glu Leu Leu Glu Val Pro Ala Glu Arg Ala Ile Met165 170 175Ile Gly Asp Ala Pro Thr Asp Leu Ala Ser Ala His Gly Ala Asp Val180 185 190Thr Ala Val Ala Ala Leu Trp Gly Cys Gln Glu Gly Ala Glu Leu Leu195 200 205Ala Ala Asp Pro Asp Val Val Leu Arg Trp Pro Ala Asp Leu Leu Ala210 215 220Leu Cys Pro Ala Leu Pro Gly His225 230<210>18<211>1071<212>DNA<213>吸水鏈霉菌(Streptomyces hygroscopicus)<220><221>CDS<222>(1)..(1071)<223>shnQ<400>18gtg aga ggc cga ata ccg gta agc atc ggc gac cgt tcg tac gag gtg48Val Arg Gly Arg Ile Pro Val Ser Ile Gly Asp Arg Ser Tyr Glu Val1 5 10 15ctg gtc ggc cgg ggg gtg cgt tcc tcg ctg gcc gaa gtg atc cag ggc96Leu Val Gly Arg Gly Val Arg Ser Ser Leu Ala Glu Val Ile Gln Gly20 25 30ctc ggc gcc cgg cgg gtc gcc gtc gtg tca gcc cgg ccc gcg gaa tgg 144Leu Gly Ala Arg Arg Val Ala Val Val Ser Ala Arg Pro Ala Glu Trp35 40 45gtg ccc gac acc ggc gtg gag acc ctg ctg ctg cct gcc cgc gac ggt 192Val Pro Asp Thr Gly Val Glu Thr Leu Leu Leu Pro Ala Arg Asp Gly50 55 60gag cgg gac aag acc ctc gcc acc gtc gag gcg ctg tgc gcg gag ttc 240Glu Arg Asp Lys Thr Leu Ala Thr Val Glu Ala Leu Cys Ala Glu Phe65 70 75 80gta cgc ttc ggc ctc acc cgc aac gac gcg gtt gtc tcc tgc ggg ggc 288Val Arg Phe Gly Leu Thr Arg Asn Asp Ala Val Val Ser Cys Gly Gly85 90 95gga acc acc acc gat gtc gtc ggc ctg gcc gcc gcc ctg tac cac cgg 336Gly Thr Thr Thr Asp Val Val Gly Leu Ala Ala Ala Leu Tyr His Arg100 105 110ggc gtg cca gtg gtg cat ctg ccg acc acg ttg ctg gcg cag gtc gac 384Gly Val Pro Val Val His Leu Pro Thr Thr Leu Leu Ala Gln Val Asp115 120 125gcg agc gtc ggc ggt aag acg gcg gtc aac ctg ccc tcc ggt aag aac 432Ala Ser Val Gly Gly Lys Thr Ala Val Asn Leu Pro Ser Gly Lys Asn130 135 140ctg gtg ggc gcc tat tgg cag cct gcc gcc gta ttg tgc gac acc gag 480Leu Val Gly Ala Tyr Trp Gln Pro Ala Ala Val Leu Cys Asp Thr Glu145 150 155 160tac ctg tcc acc ctg ccc cgg cgc gag atg ctc aac ggc ttg ggc gag 528Tyr Leu Ser Thr Leu Pro Arg Arg Glu Met Leu Asn Gly Leu Gly Glu165 170 175atc gcc cgc tgc cat ttc atc ggc gcc ggc gac ctg cgc gag ctc ggc 576Ile Ala Arg Cys His Phe Ile Gly Ala Gly Asp Leu Arg Glu Leu Gly180 185 190ctc acg gag cgt atc gcg gcc agt gtg acg ctc aag gcc ggc gtc gtc 624Leu Thr Glu Arg Ile Ala Ala Ser Val Thr Leu Lys Ala Gly Val Val195 200 205tcg gcc gat gag cgc gac acc ggt ctg cgg cac atc ctc aac tac ggc 672Ser Ala Asp Glu Arg Asp Thr Gly Leu Arg His Ile Leu Asn Tyr Gly
210 215 220cac acc ctg ggc cac gcg ctg gaa tcc gcc acc ggc ttt gcg ctt cgg720His Thr Leu Gly His Ala Leu Glu Ser Ala Thr Gly Phe Ala Leu Arg225 230 235 240cac ggc gag gcc gtc gct gtc ggc acg att ttc gcg ggc ctg ctc gcc768His Gly Glu Ala Val Ala Val Gly Thr Ile Phe Ala Gly Leu Leu Ala245 250 255ggc gct ctg gac cgg atc ggc ccc ggg cgg gtg gcg gag cac cgg gag816Gly Ala Leu Asp Arg Ile Gly Pro Gly Arg Val Ala Glu His Arg Glu260 265 270gtc gtc gag ttc tac ggc ctg ccg gcc gcg ctg ccc gag gag atc gag864Val Val Glu Phe Tyr Gly Leu Pro Ala Ala Leu Pro Glu Glu Ile Glu275 280 285acg gct gag ctg atc cgc ctg atg cgc agg gac aag aag gcg ctc acc912Thr Ala Glu Leu Ile Arg Leu Met Arg Arg Asp Lys Lys Ala Leu Thr290 295 300ggc ctc gcg ttc gtc ctc gac ggc ccc agc ggc gtg gag ctg gtc cac960Gly Leu Ala Phe Val Leu Asp Gly Pro Ser Gly Val Glu Leu Val His305 310 315 320gac gta tcc gaa cag gtc gtc gcc gat gtc ctg gag cac atg cca cgg 1008Asp Val Ser Glu Gln Val Val Ala Asp Val Leu Glu His Met Pro Arg325 330 335cag cca ctc ggc cga ctc gtc gag ccg gca cac ttc cac gga aca gga 1056Gln Pro Leu Gly Arg Leu Val Glu Pro Ala His Phe His Gly Thr Gly340 345 350gat ccg ctg gca tga 1071Asp Pro Leu Ala355<210>19<211>356<212>PRT<213>吸水鏈霉菌(Streptomyces hygroscopicus)<400>19Val Arg Gly Arg Ile Pro Val Ser Ile Gly Asp Arg Ser Tyr Glu Val1 5 10 15Leu Val Gly Arg Gly Val Arg Ser Ser Leu Ala Glu Val Ile Gln Gly20 25 30Leu Gly Ala Arg Arg Val Ala Val Val Ser Ala Arg Pro Ala Glu Trp35 40 45Val Pro Asp Thr Gly Val Glu Thr Leu Leu Leu Pro Ala Arg Asp Gly50 55 60Glu Arg Asp Lys Thr Leu Ala Thr Val Glu Ala Leu Cys Ala Glu Phe65 70 75 80Val Arg Phe Gly Leu Thr Arg Asn Asp Ala Val Val Ser Cys Gly Gly85 90 95Gly Thr Thr Thr Asp Val Val Gly Leu Ala Ala Ala Leu Tyr His Arg100 105 110Gly Val Pro Val Val His Leu Pro Thr Thr Leu Leu Ala Gln Val Asp
115 120 125Ala Ser Val Gly Gly Lys Thr Ala Val Asn Leu Pro Ser Gly Lys Asn130 135 140Leu Val Gly Ala Tyr Trp Gln Pro Ala Ala Val Leu Cys Asp Thr Glu145 150 155 160Tyr Leu Ser Thr Leu Pro Arg Arg Glu Met Leu Asn Gly Leu Gly Glu165 170 175Ile Ala Arg Cys His Phe Ile Gly Ala Gly Asp Leu Arg Glu Leu Gly180 185 190Leu Thr Glu Arg Ile Ala Ala Ser Val Thr Leu Lys Ala Gly Val Val195 200 205Ser Ala Asp Glu Arg Asp Thr Gly Leu Arg His Ile Leu Asn Tyr Gly210 215 220His Thr Leu Gly His Ala Leu Glu Ser Ala Thr Gly Phe Ala Leu Arg225 230 235 240His Gly Glu Ala Val Ala Val Gly Thr Ile Phe Ala Gly Leu Leu Ala245 250 255Gly Ala Leu Asp Arg Ile Gly Pro Gly Arg Val Ala Glu His Arg Glu260 265 270Val Val Glu Phe Tyr Gly Leu Pro Ala Ala Leu Pro Glu Glu Ile Glu275 280 285Thr Ala Glu Leu Ile Arg Leu Met Arg Arg Asp Lys Lys Ala Leu Thr290 295 300Gly Leu Ala Phe Val Leu Asp Gly Pro Ser Gly Val Glu Leu Val His305 310 315 320Asp Val Ser Glu Gln Val Val Ala Asp Val Leu Glu His Met Pro Arg325 330 335Gln Pro Leu Gly Arg Leu Val Glu Pro Ala His Phe His Gly Thr Gly340 345 350Asp Pro Leu Ala355<210>20<211>1284<212>DNA<213>吸水鏈霉菌(Streptomyces hygroscopicus)<220><221>CDS<222>(1)..(1284)<223>shnS<400>20atg tgg ggc ggt aag agc ggt gcc tac cgt gaa cgt cgc aca tcc ggc48Met Trp Gly Gly Lys Ser Gly Ala Tyr Arg Glu Arg Arg Thr Ser Gly1 5 10 15cat gtg ccc cgc ccc gct cta tgc cgt ccg tcg gcc atc ggc cga gaa96His Val Pro Arg Pro Ala Leu Cys Arg Pro Ser Ala Ile Gly Arg Glu20 25 30ata ttc gaa atg aag att tgg aga cgc atg aac gcg cga cgg aca cca 144Ile Phe Glu Met Lys Ile Trp Arg Arg Met Asn Ala Arg Arg Thr Pro
35 40 45gag ttc ccc acc tgg ccg cag tac gac gac ggc gag cgc acc ggc ctg192Glu Phe Pro Thr Trp Pro Gln Tyr Asp Asp Gly Glu Arg Thr Gly Leu50 55 60atc cgg gcc ctg gag cag ggc cag tgg tgg cgc atg gga ggc tcg gag240Ile Arg Ala Leu Glu Gln Gly Gln Trp Trp Arg Met Gly Gly Ser Glu65 70 75 80gtg gac tcc ttc gag ggt gag ttc gcg gac ttc cac ggc gcc cca cac288Val Asp Ser Phe Glu Gly Glu Phe Ala Asp Phe His Gly Ala Pro His85 90 95gct ttg gcc gtc acc aac ggc acc cac gcc ctg gag ttg gcg ttg cag336Ala Leu Ala Val Thr Asn Gly Thr His Ala Leu Glu Leu Ala Leu Gln100 105 110tgt ctg ggc gtc ggg ccg ggc acc gag gtc atc gtg ccg gcc ttc acc384Cys Leu Gly Val Gly Pro Gly Thr Glu Val Ile Val Pro Ala Phe Thr115 120 125ttc atc tcc tcc tcc cag gcc gct cag cgg ctg gga gcg gtt gcc gtc432Phe Ile Ser Ser Ser Gln Ala Ala Gln Arg Leu Gly Ala Val Ala Val130 135 140ccc gtc gac gtc gat ctc gat acc tac aac atc gac gtg gct gcc gcg480Pro Val Asp Val Asp Leu Asp Thr Tyr Asn Ile Asp Val Ala Ala Ala145 150 155 160gct tcc gcc gtc acc ccc ctc acc aag gcg atc atg cct gtg cac atg528Ala Ser Ala Val Thr Pro Leu Thr Lys Ala Ile Met Pro Val His Met165 170 175gcg ggg ctc atc gcc gac atg gac gcg ctc ggc gaa ctc tcc gcc gac576Ala Gly Leu Ile Ala Asp Met Asp Ala Leu Gly Glu Leu Ser Ala Asp180 185 190acc ggt gtg cct ctt ctc cag gac gcc gcc cac gca cac ggt gcc cgc624Thr Gly Val Pro Leu Leu Gln Asp Ala Ala His Ala His Gly Ala Arg195 200 205tgg cag ggc aaa cgg gtg ggc gag ttg ggt acg gtc gcc tcg ttc agc672Trp Gln Gly Lys Arg Val Gly Glu Leu Gly Thr Val Ala Ser Phe Ser210 215 220ttc cag aac ggc aag ctg atg acc gcc ggc gag ggc ggt gcg ctg ctc720Phe Gln Asn Gly Lys Leu Met Thr Ala Gly Glu Gly Gly Ala Leu Leu225 230 235 240ctg ccc gac gag gag acc tac gag gcc gcg ttc ctg cgg cac agt tgt768Leu Pro Asp Glu Glu Thr Tyr Glu Ala Ala Phe Leu Arg His Ser Cys245 250 255ggc cgg tca cgt acc gac cgc cga tac atg cac cag acc gcc ggc acg816Gly Arg Ser Arg Thr Asp Arg Arg Tyr Met His Gln Thr Ala Gly Thr260 265 270aac atg cgg ctc aac gag ttc tcc gcg gcc gtg ctc cgc gcc cag ctg864Asn Met Arg Leu Asn Glu Phe Ser Ala Ala Val Leu Arg Ala Gln Leu275 280 285ggc cgc ctc gac gcc cag atc acg ctc cgc gat cag cgc tgg acg ctg912Gly Arg Leu Asp Ala Gln Ile Thr Leu Arg Asp Gln Arg Trp Thr Leu
290 295 300ctg tcc cgg ctg ctc ggt gag atc gac ggc gtc gta ccc cag ggc agc960Leu Ser Arg Leu Leu Gly Glu Ile Asp Gly Val Val Pro Gln Gly Ser305 310 315 320gac ccg cgc gcc gac cgg aac tcc cac tac atg gcg atg ttc cgg atc 1008Asp Pro Arg Ala Asp Arg Asn Ser His Tyr Met Ala Met Phe Arg Ile325 330 335ccc ggc ata tcc gag gag gcc cgc aac gcc ctc gtc gac acg ctc gtc 1056Pro Gly Ile Ser Glu Glu Ala Arg Asn Ala Leu Val Asp Thr Leu Val340 345 350gag gcc ggc ctg ccc gcc ttc gcc gcc ttc cgg gcg atc tac cgc acc 1104Glu Ala Gly Leu Pro Ala Phe Ala Ala Phe Arg Ala Ile Tyr Arg Thr355 360 365gac gcg ttc tgg gag acg gcc gcg ccc gac acc acc gtc gac aag ctc 1152Asp Ala Phe Trp Glu Thr Ala Ala Pro Asp Thr Thr Val Asp Lys Leu370 375 380gcc gaa agc tgc ccg cac acc gag gcg atc agc acc gac tgc atc tgg 1200Ala Glu Ser Cys Pro His Thr Glu Ala Ile Ser Thr Asp Cys Ile Trp385 390 395 400ctg cac cat cga gtg ctg ctc gcc tcg gag gag gcc ctc cac acc aca 1248Leu His His Arg Val Leu Leu Ala Ser Glu Glu Ala Leu His Thr Thr405 410 415gcc gag atc atc gcc gac gcc gtg gcc gca cgg tga 1284Ala Glu Ile Ile Ala Asp Ala Val Ala Ala Arg420 425<210>21<211>427<212>PRT<213>吸水鏈霉菌(Streptomyces hygroscopicus)<400>21Met Trp Gly Gly Lys Ser Gly Ala Tyr Arg Glu Arg Arg Thr Ser Gly1 5 10 15His Val Pro Arg Pro Ala Leu Cys Arg Pro Ser Ala Ile Gly Arg Glu20 25 30Ile Phe Glu Met Lys Ile Trp Arg Arg Met Asn Ala Arg Arg Thr Pro35 40 45Glu Phe Pro Thr Trp Pro Gln Tyr Asp Asp Gly Glu Arg Thr Gly Leu50 55 60Ile Arg Ala Leu Glu Gln Gly Gln Trp Trp Arg Met Gly Gly Ser Glu65 70 75 80Val Asp Ser Phe Glu Gly Glu Phe Ala Asp Phe His Gly Ala Pro His85 90 95Ala Leu Ala Val Thr Asn Gly Thr His Ala Leu Glu Leu Ala Leu Gln100 105 110Cys Leu Gly Val Gly Pro Gly Thr Glu Val Ile Val Pro Ala Phe Thr115 120 125Phe Ile Ser Ser Ser Gln Ala Ala Gln Arg Leu Gly Ala Val Ala Val130 135 140Pro Val Asp Val Asp Leu Asp Thr Tyr Asn Ile Asp Val Ala Ala Ala145 150 155 160Ala Ser Ala Val Thr Pro Leu Thr Lys Ala Ile Met Pro Val His Met165 170 175Ala Gly Leu Ile Ala Asp Met Asp Ala Leu Gly Glu Leu Ser Ala Asp180 185 190Thr Gly Val Pro Leu Leu Gln Asp Ala Ala His Ala His Gly Ala Arg195 200 205Trp Gln Gly Lys Arg Val Gly Glu Leu Gly Thr Val Ala Ser Phe Ser210 215 220Phe Gln Asn Gly Lys Leu Met Thr Ala Gly Glu Gly Gly Ala Leu Leu225 230 235 240Leu Pro Asp Glu Glu Thr Tyr Glu Ala Ala Phe Leu Arg His Ser Cys245 250 255Gly Arg Ser Arg Thr Asp Arg Arg Tyr Met His Gln Thr Ala Gly Thr260 265 270Asn Met Arg Leu Asn Glu Phe Ser Ala Ala Val Leu Arg Ala Gln Leu275 280 285Gly Arg Leu Asp Ala Gln Ile Thr Leu Arg Asp Gln Arg Trp Thr Leu290 295 300Leu Ser Arg Leu Leu Gly Glu Ile Asp Gly Val Val Pro Gln Gly Ser305 310 315 320Asp Pro Arg Ala Asp Arg Asn Ser His Tyr Met Ala Met Phe Arg Ile325 330 335Pro Gly Ile Ser Glu Glu Ala Arg Asn Ala Leu Val Asp Thr Leu Val340 345 350Glu Ala Gly Leu Pro Ala Phe Ala Ala Phe Arg Ala Ile Tyr Arg Thr355 360 365Asp Ala Phe Trp Glu Thr Ala Ala Pro Asp Thr Thr Val Asp Lys Leu370 375 380Ala Glu Ser Cys Pro His Thr Glu Ala Ile Ser Thr Asp Cys Ile Trp385 390 395 400Leu His Hi s Arg Val Leu Leu Ala Ser Glu Glu Ala Leu His Thr Thr405 410 415Ala Glu Ile Ile Ala Asp Ala Val Ala Ala Arg420 42權(quán)利要求
1.利用AHBA生物合成基因保守序列篩選安莎類化合物的方法，其特征是所說的方法包括以下步驟1)AHBA生物合成基因的克??；2)吸水鏈霉菌17997中AHBA基因簇的確定3)吸水鏈霉菌17997中萘醌型安莎類化合物生物合成基因的分析；4)吸水鏈霉菌17997中萘醌型安莎霉素生物學活性測定。
2.如權(quán)利要求1所述的方法，其特征是在對NCBI數(shù)據(jù)庫中所有登錄含AHBA結(jié)構(gòu)的抗生素AHBA生物合成編碼序列分析基礎(chǔ)上，根據(jù)其氨基酸序列的保守區(qū)設(shè)計一對簡并引物上游引物為 5’-AGAGGATCCTTCGAGCRSGAGTTCGC-3’BamH1下游引物為 5’-GCAGGATCCGGAMCATSGCCATGTAG-3’以吸水鏈霉菌17997基因組DNA為模板，在LATag酶作用下進行PCR反應，獲得755bp產(chǎn)物，將其擴增產(chǎn)物與含AHBA結(jié)構(gòu)的相關(guān)抗生素進行同源性比較，確證為AHBA基因片段。
3.如權(quán)利要求1或2所述的方法，其特征是以755bpPCR產(chǎn)物為探針，選擇高嚴謹度的雜交條件與吸水鏈霉菌17997柯斯質(zhì)?；蛭膸爝M行菌落雜交及Southern雜交，獲得陽性克隆，經(jīng)測序并與NCBI數(shù)據(jù)庫進行同源性比較及保守序列分析，確定負責geldanmycin生物合成的基因簇(pCGBA10)和負責未知的萘醌型安莎類抗生素生物合成的基因簇(pCGBA3)。
4.如權(quán)利要求1或3所述的方法，其特征是通過對pCGBA3柯斯質(zhì)粒中的外源片段序列分析，確定其中含有編碼萘醌型安莎霉素生物合成的10個開放閱讀框架(ORF)，分別編碼與AHBA生物合成相關(guān)酶AHBA合酶、磷酸化酶、氧化還原酶、氨基脫氫奎尼酸合成酶及與安莎霉素合成相關(guān)酶I型聚酮合酶及酰胺合酶。
5.如權(quán)利要求1或4所述的方法，其特征是利用基因阻斷技術(shù)破壞吸水鏈霉菌17997中負責格爾德霉素生物合成基因，以檢測該菌產(chǎn)生的另外一個萘醌類安莎霉素的生物學活性，具體步驟是將吸水鏈霉菌17997中與格爾德霉素生物合成相關(guān)序列插入質(zhì)粒載體如噬菌體載體KC515，構(gòu)建重組質(zhì)粒，轉(zhuǎn)染17997孢子，獲得溶源菌，分離提取溶源菌總DNA，用限制性酶(如BamHI)酶切后，以載體上標記如KC515載體上硫鏈絲菌素抗性標記基因為探針，進行Southern雜交分析，證明溶源菌中所含與格爾德霉素生物合成相關(guān)基因已和載體一起整合至染色體，致使格爾德霉素生物合成基因受到破壞，以單純皰疹病毒1型感染的VERO細胞為模型，測定溶源菌發(fā)酵液對病毒的活性。
6.利用AHBA生物合成基因保守序列，篩選安莎類化合物的方法的思路，在篩選其它類有生物學活性化合物中的應用。
全文摘要
本發(fā)明涉及一種利用參與微生物產(chǎn)物生物合成酶編碼基因保守序列從微生物中鑒定尋找新的活性物質(zhì)的方法，具體來講，就是利用分子生物學手段，根據(jù)安莎類化合物特異結(jié)構(gòu)的AHBA生物合成基因保守序列，克隆相關(guān)基因，并以此為探針，獲得安莎類化合物生物合成基因簇和鑒定新的安莎類抗生素，該方法準確度高，效果顯著，并可用于高通量篩選。
文檔編號C12Q1/68GK1398986SQ02125509
公開日2003年2月26日 申請日期2002年7月17日 優(yōu)先權(quán)日2002年7月17日
發(fā)明者王以光, 高群杰 申請人:中國醫(yī)學科學院醫(yī)藥生物技術(shù)研究所