一种改进的用高拷贝质粒制备BAC载体的新方法  

An Improved Method for the Preparation of BAC Vector DNA with High-copy Plasmid

作　　者：邓代永[1] 杨继良[1] 宣劲松[1] 张明哲[1] 翁曼丽[1] 王斌[1] 

机构地区：[1]中国科学院遗传与发育生物学研究所,北京100101

出　　处：《高技术通讯》2003年第7期29-33,共5页Chinese High Technology Letters

基　　金：863计划 ( 2 0 0 1AA62 10 90 ); 973规划 ( 2 0 0 1CB10 880 2 )资助项目

摘　　要：介绍了一种改进的使用高拷贝质粒制备BAC载体的简便有效的方法 ,我们同时使用从高拷贝质粒 pCUGIBAC1制备的和用传统方法从单拷贝质粒 pIndigoBAC 5制备的两种载体 ,比较了这两种载体与玉米大片段DNA连接所构建的BAC文库。结果表明在使用这两种BAC载体构建的BAC文库中 ,所获得的转化体数目 ,及在BAC克隆中插入的DNA片段的大小和BAC克隆的稳定性等方面都相同 ,从而证明本文所介绍的从高拷贝质粒 pCUGIBAC1来制备BAC载体是一种更方便的方法 。Here we introduce an improved method for preparing BAC vector with high copy plasmid pCUGIBAC1 In this paper two kinds of vector, the vector pIndigoBac536 from high copy plasmid pCUGIBAC1 and the vector pIndigoBAC 5 from a traditional single copy plasmid, have been comparatively used to construct a maize BAC library. The results show that there is no apparent difference between them in the efficiency of generation transformants, in the inserted DNA fragment size for BAC clones and in the stability of BAC clones in E. coli . Therefore, it can be safely concluded that the improved method for preparing BAC vector from high copy plasmid pCUGIBAC1 is a convenient, time saving and high efficient method, and will significantly facilitate the construction of BAC library.

关 键 词：高拷贝质粒 细菌人工染色体 载体 制备方法 DNA 转化体 克隆稳定性 BAC文库 

分 类 号：Q78[生物学—分子生物学]