黑暗链霉菌tobS1-C基因缺失突变株的构建  

Construction of Streptomyces tenebrarius mutant with knockout of tobS1-C genes

作　　者：林玉双 洪文荣[2] 

机构地区：[1]常州方圆制药有限公司,常州213022 [2]福州大学生物科学与工程学院,福州350108

出　　处：《中国药科大学学报》2012年第1期92-96,共5页Journal of China Pharmaceutical University

基　　金：国家自然科学基金资助项目(No.31070093);国家"重大新药创制"科技重大专项资助项目(No.2012ZX09201-101-008)~~

摘　　要：通过敲除妥布霉素生物合成基因tobS1-C,定向选育阿泊拉霉素高产菌株。PCR扩增tobS1-C基因上下游同源序列和红霉素抗性基因ermE,构建穿梭载体pSH6,利用接合转移法转化黑暗链霉菌Tt49,经同源重组,敲除tobS1-C两基因序列,共2 484 bp,获得框内删除突变株黑暗链霉菌T103(△tobS1C)。发酵验证表明突变株不再合成妥布霉素,只合成阿泊拉霉素。In order to breed a mutant strain for high production of apramycin,carbamoyltobramycin biosynthesis genes tobS1-C were delected from Streptomyces tenebrarius through homologous recombination.Gene deletion vector pSH6 was constructed and introduced into S.tenebrarius Tt49 through conjugal transfer.TobS1-C genes in the chromosome were displaced by deletion allele on the plasmid via double crossover.Then the mutant S.tenebrarius T103（△tobS1C）was proved to be genetically stable.Shaking flask experiments and TLC analysis showed that the mutant strain only produced apramycin.

关 键 词：黑暗链霉菌 阿泊拉霉素 同源重组 基因缺失 

分 类 号：Q784[生物学—分子生物学]