链霉菌Streptomyces tenebrarius H6中与抗生素有关的糖生物合成基因的克隆  

Cloning of the Sugar Related Biosynthesis Gene Cluster from Streptomyces tenebrarius H6

作　　者：李天伯[1] 尚广东[2] 夏焕章[1] 王以光[2] 

机构地区：[1]沈阳药科大学,沈阳110015 [2]中国医学科学院中国协和医科大学医药生物技术研究所,北京100050

出　　处：《生物工程学报》2001年第3期329-331,共3页Chinese Journal of Biotechnology

基　　金：国家自然科学基金面上项目 !( 3970 0 0 0 3)&&

摘　　要：Streptomyces tenebrarius H6 produces a complex of aminoglycoside antibiotics,such as apramycin,tobramycin and kanamycin B etc. To study the apramycin biosynthetic genes the genomic library from the Streptomyces tenebrarius H6 was established using E.coli/Streptomyces shuttle vector pKC505 by in vitro packing.The probability of finding a specific gene from the library composed of 3 000 colonies was over 99 9%. According to the highly conserved sequence of the genes involved in 6\|deoxyhexose biosynthesis,primers were designed and 0 6kb fragment homologous to strE gene was obtained by PCR.30 positive clones were found from the genomic library of \%S.tenebrarius\% H6 with the 0 6kb fragment as a probe.Overlapped regions were localized by Southern hybridization and putative sugar related biosynthetic gene cluster was mapped by restriction enzyme digestions.An ORF of dTDP\|glucose\|4,6\|dehydratase gene consisted of 1,132bp,designated as apr E,was obtained and submitted to GenBank under the accession number of AF306787.A DNA sequence highly homologous to str L coding dTDP\|4\|dehydrorhamnose reductase was found linked with apr E gene.Streptomyces tenebrarius H6 produces a complex of aminoglycoside antibiotics,such as apramycin,tobramycin and kanamycin B etc. To study the apramycin biosynthetic genes the genomic library from the Streptomyces tenebrarius H6 was established using E.coli/Streptomyces shuttle vector pKC505 by in vitro packing.The probability of finding a specific gene from the library composed of 3 000 colonies was over 99 9%. According to the highly conserved sequence of the genes involved in 6\|deoxyhexose biosynthesis,primers were designed and 0 6kb fragment homologous to strE gene was obtained by PCR.30 positive clones were found from the genomic library of \%S.tenebrarius\% H6 with the 0 6kb fragment as a probe.Overlapped regions were localized by Southern hybridization and putative sugar related biosynthetic gene cluster was mapped by restriction enzyme digestions.An ORF of dTDP\|glucose\|4,6\|dehydratase gene consisted of 1,132bp,designated as apr E,was obtained and submitted to GenBank under the accession number of AF306787.A DNA sequence highly homologous to str L coding dTDP\|4\|dehydrorhamnose reductase was found linked with apr E gene.

关 键 词：黑暗链霉菌 dTDP-葡萄糖-4 6-脱水酶基因 糖生物合成基因 抗生素 

分 类 号：Q786[生物学—分子生物学]