DNA改组技术在水蛭素实验进化中的应用  

Directed Evolution of Hirudin by DNA shuffling

作　　者：周育[1] 刘晖[1] 谭涛超[1] 黄仪秀[1] 朱圣庚[1] 

机构地区：[1]北京大学生命科学学院,北京100871

出　　处：《中国生物化学与分子生物学报》2001年第2期271-274,共4页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家自然科学基金! (No.395 80 0 16 );博士学科点专项科研基金&&

摘　　要：Hirudin gene was mutated by error\|prone PCR and DNA shuffling.The shuffled genes were inserted into phagemid pCANTAB\|5G8.After the introduction of the recombinant DNA into E.coli TG1,the hirudin mutants were displayed on the surface of bacteriophage M13.A mutant with increased specific activity of antithrombin was selected by two rounds of panning with decreased amounts of thrombin,and was identified as N47K.Activity analysis of HV2 and its mutants N47K,N26S,D5A,P46L/S50G suggested that K47,P46 were important to the interaction of hirudin and thrombin.Hirudin mutants with improved properties could be hopefully aquired through directed evolution by DNA shuffling.Hirudin gene was mutated by error\|prone PCR and DNA shuffling.The shuffled genes were inserted into phagemid pCANTAB\|5G8.After the introduction of the recombinant DNA into E.coli TG1,the hirudin mutants were displayed on the surface of bacteriophage M13.A mutant with increased specific activity of antithrombin was selected by two rounds of panning with decreased amounts of thrombin,and was identified as N47K.Activity analysis of HV2 and its mutants N47K,N26S,D5A,P46L/S50G suggested that K47,P46 were important to the interaction of hirudin and thrombin.Hirudin mutants with improved properties could be hopefully aquired through directed evolution by DNA shuffling.

关 键 词：DNA改组技术 应用 水蛭素 实验进化 应用 

分 类 号：Q784[生物学—分子生物学] R973.2[医药卫生—药品]