牛结核分枝杆菌基因文库的构建  

Construction of DNA Library of M·bovis TMC410

作　　者：蔡宏[1,2,3] 陈东[1,2,3] 李淑霞[1,2,3] 呼西旦 李君莲[1,2,3] 陈永福 

机构地区：[1]新疆畜牧科学院兽医研究所 [2]中国农业大学农业生物技术实验室 [3]新疆结核病研究所

出　　处：《中国预防兽医学报》1999年第2期114-116,共3页Chinese Journal of Preventive Veterinary Medicine

摘　　要：以国际标准牛型结核分枝杆菌（Ｍ．ｂｏｖｉｓ）ＴＭＣ４１０为材料，制备其总ＤＮＡ，经过限制性内切酶Ｓａｕ３Ａ部分消化后，纯化回收９～２５Ｋｂ片段，并将其用Ｔ４ＤＮＡ连接酶与经ＢａｍＨＩ酶切为粘性末端的ＬａｍｂｄａＥＭＢＬ３左右臂连接。用噬菌体包装蛋白包装，再进行连续稀释，然后将不同稀释度的噬菌体转移到宿主菌ＬＥ３９２。所获重组子为６×１０４，因为牛结核杆菌基因组大小约为４×１０６Ｋｂ则以９９％概率筛到任一基因，要求的重组子数为１．０８×１０３。Total DNA was partially digested with Sau3A and 9～25Kb DNA fragments were recovered form agarose gel.The products were ligated with EMBL3 arms by T 4 DNA ligase and packaged using an in vitro packaging extract.The bacteriophages with different dilution were used to transfect bacteria LE392.The number of bacterial recombinants obtained was about 6×10 4.The genomic DNA Library Constructed could provide sufficient clones to cover all M.bovis gene.

关 键 词：牛 结核分枝杆菌 基因文库 

分 类 号：S852.61[农业科学—基础兽医学] S858.235.1[农业科学—兽医学]