血清Ⅱ型鸭疫里默氏杆菌基因组文库的构建  

CONSTRUCTION OF GENOMIC LIBRARY OF RIEMERELLA ANATIPESTIFER SEROTYPE Ⅱ

作　　者：林树乾[1] 何元龙[1] 赵增成[1] 李峰[1] 

机构地区：[1]山东省农业科学院家禽研究所,济南250023

出　　处：《中国动物传染病学报》2010年第6期38-42,共5页Chinese Journal of Animal Infectious Diseases

基　　金：山东省科学技术发展计划(2008GG30008026);高校院所自主创新计划(201004029)

摘　　要：为研究血清Ⅱ型鸭疫里默氏杆菌的抗原基因,筛选毒力较强的血清Ⅱ型鸭疫里默氏杆菌菌株,提取其基因组,分别用Sau3A I酶切法和shotgun法对基因组进行处理后,选取1.5～3 kb的DNA片段,与PUC118载体连接后电转化JM109大肠杆菌感受态细胞,构建血清Ⅱ型鸭疫里默氏杆菌基因组文库。两种方法得到的库容量分别为40 000和50 000左右,随机挑选阳性克隆鉴定表明有外源DNA片段的插入,说明建库成功。该文库的成功构建为Ⅱ型鸭疫里默氏杆菌新的抗原基因的筛选奠定了基础。The genomic library of a virulent Riemerella anatipestifer serotype Ⅱ was constructed to identify the protective antigens.The genomic DNA was digested with restriction endonuclease Sau3A Ⅰ and cut into pieces by shotgun method.These DNA fragments with the size from 1.5 to 3 kb were ligated into the plasmid vector PUC118.Then,the recombinant plasmid was transformed into E.coli JM109 by electroporation to construct the genomic library with the capability about 40000 and 50000.The insertion of exotic DNA fragments was confirmed by PCR that amplified positive clones from random selected samples.

关 键 词：鸭疫里默氏杆菌 基因文库 构建 

分 类 号：S858.322.618[农业科学—临床兽医学] Q785[农业科学—兽医学]