细菌人工染色体基因组文库构建关键技术研究  

Discuss of Several Problems in Bacterial Artificial Chromosome Library Construction

作　　者：刘长青[1] 吴宏梅[1] 包阿东[1] 陆涛峰[1] 刘帅[1] 张洪海[2] 唐学玺[1] 关伟军[1] 马月辉[1] 

机构地区：[1]中国农业科学院北京畜牧兽医研究所 [2]曲阜师范大学研究生处,曲阜273165

出　　处：《生物技术通报》2008年第4期66-69,共4页Biotechnology Bulletin

基　　金：国家“863”计划项目资助(2006AA10Z198)(2007AA10Z170)

摘　　要：基因组文库是进行分子克隆和基因结构与功能研究的基础,完整覆盖的基因组文库的构建,使基因组任何DNA片段的筛选和获得成为可能。细菌人工染色体(Bacterial Artificial Chromosome,BAC)与其它载体系统相比具有转化效率高、嵌合体少、插入片段易回收,高覆盖率、稳定性强,并且具有易分离和操作等特性等优点而被广泛应用。作为物种保护策略的重要部分,构建我国濒危动植物品种基因组BAC文库,对于其遗传资源的保护和研究具有非常重要的作用。在查阅大量国内外相关资料的基础上,就BAC基因组文库构建过程中载体的制备、高分子量DNA的制备、大片段DNA的回收、连接与电击转化、基因组BAC文库质量鉴定等几个重点、难点问题进行了详细的论述和分析,对于构建高分子量插入片段、高覆盖率和稳定性的基因组文库提供理论基础与技术支撑。The construction of a genomic library is essential to research on genome structure and function. A comprehensive genomic library makes it possible to screen and isolate any genomic DNA fragments from it. Artificial bacterial chromosome can hold large segment DNA,which have higher coverage ratio and stability than other vectors in the construction of genome library. It is also easy to separate and manipulate. Some problems of the construction of genome library about preparation of BAC vector,isolation of high molecular weight nuclear DNA,isolation of size-selected DNA from agarose,more coverage ratio and stability were discussed,thus being able to supply a basis for the genome library of large-size insert,more coverage and stability.

关 键 词：细菌人工染色体 基因组文库构建 质粒纯化 高分子量DNA 

分 类 号：Q78[生物学—分子生物学] Q785