聚合酶链式反应递缩基因组文库以克隆抗生素生物合成基因簇  

Cloning of Antibiotic Biosynthetic Gene Clusters from Genomic Library by Narrow-down Polymerase Chain Reaction

作　　者：何云龙[1] 朱冬青[1] 白林泉[1] 邓子新[1] 

机构地区：[1]上海交通大学生命科学与技术学院微生物代谢教育部重点试验室,上海200240

出　　处：《上海交通大学学报》2009年第1期5-8,共4页Journal of Shanghai Jiaotong University

基　　金：国家自然科学基金资助项目(30570019)

摘　　要：在克隆阿维菌素(avermectin)生物合成基因簇的过程中,探索出了一条运用聚合酶链式反应(PCR)扩增技术对链霉菌基因组文库进行递缩法筛选、以克隆抗生素生物合成基因簇的新方法.依次以96孔培养板整板混合质粒、单排混合质粒和单个菌落为模板进行PCR扩增,以逐步递缩的方式定位包含目的基因的克隆.结果表明,与传统的菌落原位杂交方法相比,该方法具有简单、快捷、准确率高等优点.A new strategy of narrowing down antibiotic biosynthetic gene cluster from genomic library by polymerase chain reaction （PCR） was carried out when cloning avermectin biosynthetic gene cluster. Cosraids containing targeting gene were identified by narrow-down PCR using the mix DNA of 96 cosmids in one plate, the mix DNA of 12 cosmids in each line and the single strain as template separately. Compared with traditional method of in situ colony hybridization, this method is easier, faster and more veracious.

关 键 词：聚合酶链式反应 阿维菌素 克隆 基因组文库 基因簇 递缩法 

分 类 号：Q781[生物学—分子生物学]