优化菌落原位杂交体系筛选野生稻基因组文库  

Optimized Colony in situ Hybridization System for Screening Wild Rice Genomic Library

作　　者：陶小平 李磊[1,2] 李强[1,2] 韩小霞 邢俊杰[1,2,3] 崔玲玲[1,2] 李玲珑[1,2] 张朝良 张秋平 曹孟良[1,2,3] 

机构地区：[1]湖南长沙西城杂交水稻基因工程研究中心,中国湖南长沙410205 [2]中南大学生物科学与技术学院,中国湖南长沙410083 [3]国家杂交水稻工程技术研究中心,中国湖南长沙410205

出　　处：《生命科学研究》2008年第4期328-332,共5页Life Science Research

基　　金：973项目(2007CB109007);863重点项目(2006AA100101);国家科技支撑计划(2007BAD77B00);中国博士后基金(2005037696);湖南省自然科学基金(07JJ3029)资助项目

摘　　要：菌落原位杂交仍是当今筛选基因组文库的重要方法之一,但难以保持背景清晰和阳性杂交点明显的稳定的杂交体系.通过对杂交膜不同处理方法的对比,表明利用溶菌酶处理杂交体系进行菌落原位杂交筛选,其杂交结果背景较干净,且能根据杂交信号的强弱很清晰地显色阳性黑点.采用该法筛选野生稻基因组文库,已筛选出22个阳性克隆,并测序分析均为阳性克隆.Colony in situ hybridization is still a very important method to screen genome library today. However, it is hardly to keep a stable hybridization system to obtain a clear background and the obvious positive hybridization points. The results of comparing different treatments for hybridization indicated that by using lysozyme treatment system for colony in situ hybridization, the background of hybridization is clearer and positive dark dots can be distinguished clearly according to the hybridization signal intensity. 22 positive clones have been screened by using this method from wild rice genomic library and confirmed by sequencing analysis.

关 键 词：菌落原位杂交 基因组文库 溶菌酶 阳性克隆 

分 类 号：Q75[生物学—分子生物学]