泰乐菌素基因工程菌的构建  

Construction of a gene-engineered strain for the production of tylosin

作　　者：范亮[1] 杜宝华[1] 陈光[1] 田威[1] 何建勇[1] 

机构地区：[1]沈阳药科大学生命科学与生物制药学院,辽宁沈阳110016

出　　处：《沈阳药科大学学报》2012年第5期398-401,共4页Journal of Shenyang Pharmaceutical University

摘　　要：目的构建具有双拷贝tylF基因的泰乐菌素基因工程菌,以解决泰乐菌素基因生物合成的限速环节,提高泰乐菌素的发酵产量。方法通过SOE-PCR获得PermE-tylF基因,构建随机整合型重组质粒pBH05,通过接合转移将PermE-tylF基因及其载体随机整合到弗氏链霉菌的基因组中,并通过抗性筛选获得重组菌株D-120。将重组菌株D-120进行摇瓶发酵,采用生物效价测定的二剂量法测定泰乐菌素的发酵单位。结果在相同的发酵条件下,重组菌株泰乐菌素的发酵单位较出发菌株提高32.7%。结论该基因工程菌的构建改善了泰乐菌素生物合成的限速环节,大幅度提高了泰乐菌素的发酵效价。Objective To construct a gene-engineered strain which producing tylosin by introducing PermE-tylF gene in the genomic DNA.Methods PermE-tylF gene was obtained by SOE-PCR.An E.coli-Streptomyces shuttle plasmid pBH05 containing PermE-tylF gene was constructed and was introduced into Streptomyces fradiae Ty-2010 through conjugal transfer.Tylosin fermentation was conducted by shaking flask method and tylosin titer was determined by bacterial inhibition tests.Results The tylosin productivity of recombinant strain D-120 was increased by 32.7% than the original strain.Conclusions The productivity of tylosin is obviously increased by the gene-engineered strain,in which the copy number of tylF gene is increased by introducing PermE-tylF gene in the genomic DNA of Streptomyces fradiae.

关 键 词：弗氏链霉菌 tylF基因 泰乐菌素 基因工程菌 

分 类 号：R978.1[医药卫生—药品]