小麦BAC shotgun测序文库的构建  

Construction of Wheat BAC Shotgun Library

作　　者：张俊成[1] 孔秀英[2] 吴佳洁[2] 刘越[2] 张春庆[1] 

机构地区：[1]山东农业大学农学院,山东泰安271018 [2]农业部作物种质资源与生物技术重点开放实验室/中国农业科学院作物科学研究所/国家农作物基因资源与基因改良重大科学工程,北京100081

出　　处：《麦类作物学报》2007年第3期374-377,406,共5页Journal of Triticeae Crops

基　　金：中澳合作项目(GRDCgrantET5)

摘　　要：为给小麦基因组中某一基因区域的测序与分析奠定基础,以小麦7B和7D染色体上的10个BAC克隆为材料,运用Large-Construct Kit和TOPO TA cloning kit两个商品化的试剂盒及HydroShearDNA剪切仪建立了BAC shotgun文库构建的技术体系,即利用该体系可以获得高纯度的BAC 20-50μg以及DNA 3-5 kb的目标小片段,5-20 min即可完成连接,一次转化可获得2000个重组克隆。利用这一方法,现已构建了10个小麦BAC shotgun文库,这些文库可为小麦基因组特定区段的研究奠定基础。BAC shotgun library is an important platform for sequencing the whole genome or some specific regions. The objective of this study is to establish the construction system of BAC shotgun library and service the analysis of specific regions wheat. Ten BACs from 7B and 7D of wheat were used as materials. Two commercialized kits of Large-Construct Kit and TOPO TA cloning kit, and DNA shearing instrument-HydroShear were employed in the construction of BAC shotgun library. By using the established system, 20-50μg highly purified DNA for BAC could be extracted quickly and easily; the ligation reaction could be finished within 5 - 20 minutes and 2 000 recombinant clones could be produced through one transformation. Using this method, we have constructed 10 wheat BAC libraries. These BAC shotgun libraries will provide a platform for gene cloning and wheat genome research.

关 键 词：小麦 BAC克隆 shotgun文库 

分 类 号：S512.1[农业科学—作物学] S336