Construction of mini-Tn4001 transposon vector for Mycoplasma gallisepticum.
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.
Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, China.
HongJun
H
Chen
ChunMei
C
Zhao
XieYue
X
Shen
DanQing
D
Chen
ShengQing
S
Yu
Chan
C
Ding
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D004251:DNA Transposable Elements; D005822:Genetic Vectors; D045686:Mycoplasma gallisepticum / Q000235:genetics* / Q000472:pathogenicity; D014169:Transformation, Bacterial; D019895:Transposases / Q000235:genetics
D001426:Bacterial Proteins; D004251:DNA Transposable Elements; D019895:Transposases