Construction of mini-Tn4001 transposon vector for Mycoplasma gallisepticum

作     者：CHEN HongJun, ZHAO ChunMei, SHEN XieYue, CHEN DanQing, YU ShengQing & DING Chan Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, China 

作者机构：Shanghai Veterinary Research Institute Chinese Academy of Agricultural Sciences Shanghai China 

出 版 物：《Science China(Life Sciences)》 (中国科学（生命科学英文版）)

年 卷 期：2010年第53卷第11期

页      面：1340-1345页

核心收录：

学科分类：0710[理学-生物学] 090601[农学-基础兽医学] 0830[工学-环境科学与工程（可授工学、理学、农学学位）] 09[农学] 0906[农学-兽医学] 

基　　金：supported by the National Natural Science Foundation of China (Grant Nos.30871883 and 31001077) the National Basic Foundation of China (Grant Nos.2006JB01 and 2008JB13) the Shanghai Agri & Tech Foundation (Grant No.2007-11-2) 

主　　题：Mycoplasma gallisepticum mini-Tn4001 transposon vector mycoplasma transformation 

摘      要：The detailed genetic analysis of mycoplasmas has long been hampered by the lack of appropriate tools for genetic manipulation. In this study, the transposon vector, mini-Tn4001tetM, was constructed containing the tnp gene, encoding a transposase gene in Staphylococcus aureus, two copies of the IS256 inverted repeat sequence (inner and outer) and the tetM gene, from the Enterococcus faecalis Tn916 transposon, conferring resistance to tetracycline. This vector was electro-transformed into Mycoplasma gallisepticum (MG). The recombinant cells were screened by tetracycline selection. The results indicated that the transposon vector could replicate in MG strain R by successive passages, indicating that MG is a potential vector for expressing protective antigens of other pathogens.