Streptomyces lincolnensis新遗传操作方法的建立以及lmbQ基因功能研究  

Construction of the New Genetic Manipulation Method of Streptomyces lincolnensis and lmbQ Gene Function Verification

作　　者：王增亮[1] 赵群飞[2] 高淑红[1] 陈长华[1] 刘文[2] 

机构地区：[1]华东理工大学生物工程学院,上海200237 [2]中国科学院上海有机化学研究所,上海200032

出　　处：《生物技术通报》2010年第11期186-190,198,共6页Biotechnology Bulletin

摘　　要：以载体pSET152和pKC1139为出发质粒,通过供体菌大肠杆菌ET12567和S17-1转入林可链霉菌(Streptomyces lincolnensis),建立和优化了接合转移体系。林可霉素生物合成基因簇中,lmbQ基因可能编码一个调控蛋白,构建了S.lin-colnensislmbQ基因中断载体,通过接合转移导入S.lincolnensis野生型菌株,筛选得到基因双交换突变株,通过PCR以及测序验证基因型正确,该突变株即为lmbQ同框敲除突变株。摇瓶发酵结果表明,lmbQ基因是一个正调控基因。Intergeneric transfer of plasmid vectors pSET152 and pKC1139 from donor strain Escherichia coli ET12567 or S17-1 to Streptomyces lincolnensis was constructed and optimized.The lmbQ gene in the lincomycin gene cluster encode a putative regulatory protein.We constructed the gene disruption vector and then transformed into S.lincolnensis through inter-species conjugation.After PCR amplification and sequencing,we confirmed the resulting double cross-over strains are the lmbQ ＂In-frame deletion＂ mutants.The fermentation at the flask level proved that the lmbQ is a positive regulation gene.

关 键 词：林可链霉菌 林可霉素 属间接合转移 lmbQ基因 同框敲除 

分 类 号：Q78[生物学—分子生物学]