大豆过氧化物酶基因随机突变文库的构建  

Construction of Soybean Peroxidase Gene Random Mutation Library

作　　者：曾家豫[1] 郑群[1] 廖世奇[2] 王黎[2] 袁红霞[2] 杨海棠[1] 张丽琼[3] 张宁[3] 

机构地区：[1]西北师范大学生命科学学院,甘肃兰州730070 [2]甘肃省医学科学研究院,甘肃兰州730050 [3]兰州理工大学生命科学与工程学院,甘肃兰州730050

出　　处：《大豆科学》2011年第5期731-737,共7页Soybean Science

基　　金：甘肃省教育厅资助项目(0601-28,0501B-16);甘肃省自然科学基金资助项目(096RJZA035);甘肃省高分子材料重点实验室开放课题资助项目(KF-06-01)

摘　　要：为提高大豆过氧化物酶(soybean peroxidase,SBP)的活性及研究该酶一级结构与酶活性间的关系,采用连续易错PCR方法对大豆过氧化物酶基因(sbp)进行了2轮随机突变,将第二轮易错PCR产物与质粒载体pPICZα-A分别进行双酶切,经T4DNA连接酶催化连接后,CaCl2法转化重组质粒pPICZα-A-sbp至大肠杆菌DH5α中,成功构建出sbp随机突变文库,库容量约为1.77×106 cfu。In order to improve enzymatic activity of soybean peroxidase（SBP）and study the correlation between its primary structure and enzymatic activity,sequential error-prone PCR method was used on soybean peroxidase genes（sbp） for two times randon mutation.The second round error-prone PCR products and plasmid vectors pPICZα-A were digested by double enzymes.After the PCR products ligated into the vectors by T4DNA ligase,recombinant plasmids pPICZα-A-sbp were transformed into Escherichia coli DH5α through calcium chlorid method.A random mutation library was constructed,and the capacity was about 1.77×106 cfu.

关 键 词：大豆过氧化物酶 易错PCR 基因文库 随机突变 酶定向进化 

分 类 号：S565.1[农业科学—作物学]