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.
出处
《生物技术通报》
CAS
CSCD
北大核心
2010年第11期186-190,198,共6页
Biotechnology Bulletin
