绒山羊细菌人工染色体BAC文库构建条件的优化  

Optimizing the Constructing Condition of Bacterial Artificial Chromosome Library in the Cashmere Goat

作　　者：刘志红[1] 李宁[2] 任立明[2] 胡晓湘[2] 郭英[2] 崔文涛[3] 尹俊[1] 张文广[1] 李金泉[1] 

机构地区：[1]内蒙古农业大学动物科学与医学学院,呼和浩特010018 [2]中国农业大学农业生物技术国家重点实验室,北京100193 [3]中国农业科学院北京畜牧兽医研究所,北京100193

出　　处：《中国畜牧兽医》2009年第10期55-58,共4页China Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金地区联合资助项目(30660122);"863"计划项目(2007AA10Z151);国家科技支撑计划(2006BAD14B07;2006BAD01A11;2007BAD-56B03);国家自然科学基金项目(30760163)

摘　　要：近年来,细菌人工染色体BAC文库成为应用最广泛的基因组文库之一,它以其容量大、遗传特性稳定、嵌合体少、插入片段易回收、操作简便等不可比拟的优点而被广泛应用于基因组较大的真核生物基因组研究之中。作者就细菌人工染色体BAC文库的构建条件进行了研究,用HindⅢ部分酶切绒山羊基因组,脉冲场凝胶电泳分离大片段DNA,以电洗脱法回收DNA后,与BAC载体连接,将连接产物点透析后,电转化DH10B感受态细胞,最后对转化的片段小提质粒进行NotⅠ酶切,脉冲电泳鉴定其大小。结果表明,本试验采用的方法既可获得较大的插入片段又可获得较高的转化效率,是较好的用于构建大基因组文库的方法。In recent years, bacterial artificial chromosome （BAC） library has become one of the most popular used genome library, and it is used generally in the research of eukaryotic organism that has longer genome by its advancement including lar ger capacity, stable inherited characteristic, few chimera, easily reclaiming insertion element, convenient operation and so on. This article researched the constructing condition of BAC library, cutting the goat＇s chromosome by HindⅢenzyme, separating the big DNA fragment by pulsed field gel eleetrophoresis, reclaiming DNA by eleetrodialysis, linking with BAC bearer taken off phosphorous, transforming competent cell DH10B by elutriations, and finally evaluating the length of fragment by cutting the DNA with Not I enzyme. The consequence indicated that the method that this research chose can get not only the longer insertion element, but also the higher converting efficiency, and it is the better method to construct the bigger genome BAC library.

关 键 词：细菌人工染色体 文库 构建条件 

分 类 号：S827.2[农业科学—畜牧学]