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Expression vector construction and prokaryotic expression of
TM1 gene of Mycoplasma gallisepticum H3 strain
HAO Yongqing1, WANG Xiuqing1, ZHOU
Yanjun2,
TONG Guangzhi2, GAO
Jinliang3, ZHAO Zhenhua1, WU
Ni1
(1. College of Animal Science and Veterinary Medicine, Nei Monggol Agricultural University,
Hohhot 010018, China; 2. Harbin Veterinary Research Institute, Chinese Academy of
Agricultural Sciences, Harbin 150001, China; 3. Lanzhou Veterinary Research
Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China) |
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Abstract: The abstracted recombinant plasmid of TM1 gene of Mycoplasma gallisepticum (MG) H3 strain was digested with Eco RⅠ and Sal Ⅰ. The obtained fragment
786bp in length was inserted into the prokaryotic expression vector pET30a1 digested by the same restriction endonucleases and transformed into BL21（DE3）. The insert position, size and open reading frame (ORF) were all confirmed to be right by PCR, restriction digestion and sequencing analysis. Thus the prokaryotic expression vector was constructed successfully. Positive clone was induced with IPTG for expression of TM1 gene. SDSPAGE analysis showed that molecular weight of the expressed protein was
29ku. Westernblotting result showed that the recombinant protein can be recognized by positive serum of MG.
Key words: Mycoplasma gallisepticum ； TM1 gene； expression vector construction； prokaryotic expression |
