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 

出　　处：《Science China(Life Sciences)》2010年第11期1340-1345,共6页中国科学（生命科学英文版）

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)

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.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.

关 键 词：MYCOPLASMA gallisepticum mini-Tn4001 TRANSPOSON VECTOR MYCOPLASMA TRANSFORMATION 

分 类 号：S852.62[农业科学—基础兽医学]