石麦15细菌人工染色体文库构建与鉴定  

Construction and Characterization of BAC Library from Common Wheat Shimai 15

作　　者：李孟军[1] 杨煜[2] 高欣娜[1] 郭晓[2] 何明琦[1] 史占良[1] 郭进考[1] 

机构地区：[1]石家庄市农林科学研究院/河北省小麦工程技术研究中心,河北石家庄050041 [2]山东省农业科学院蔬菜花卉研究所,山东济南250100

出　　处：《麦类作物学报》2014年第2期164-168,共5页Journal of Triticeae Crops

基　　金：国家现代农业产业技术体系项目(CARS-3-1);石家庄市科技支撑计划项目(12149402A)

摘　　要：为给小麦功能基因研究提供参考信息,以冬小麦栽培品种石麦15（Triticum aestivum L.）为试材,从暗培养7~10d的黄化幼苗地上部提取高分子量基因组DNA,使用载体pCC1BAC的HindⅢ位点构建了六倍体小麦细菌人工染色体文库。该文库由1 000 000个以上克隆组成,保存在1 020个混合池中,克隆插入片段平均大小为85kb,空载率为2.5%;覆盖小麦基因组5倍以上;挑取7个克隆培养5d后,经HindⅢ完全酶切检测,其指纹图谱稳定一致。石麦15细菌人工染色体文库的构建为小麦基因克隆、调控序列克隆以及比较基因组学研究奠定了基础。A bacterial artificial chromosome library of hexaploid wheat was constructed using the win- ter wheat （Triticum aestivum L. ）cultivar ＇Shimai 15＇. Fresh shoot tissue from 7- 10 day-old seedlings was used to isolate HMW genomic DNA. The library was constructed using the HindIlI site of pCCIBAC vector（Hind Ⅲ ）. The whole library contained more than 1 000 000 BAC clones and preserved in 1020 pools. The average insert size of BAC clones is about 85 kb and the empty clones are about 2.5 %. The library represents common wheat genome coverage of five fold. Seven clones picked randomly from this library showed no Hind Ⅲ fingerprint changes after cultured 5 days, which indicates that the BAC clones are stable in the library. The library lays the foundation for gene cloning, regulatory-sequence cloning and comparative genomics research of common wheat.

关 键 词：小麦 细菌人工染色体文库 插入片段 稳定性 

分 类 号：S512.1[农业科学—作物学] S330