Journal Article
Cloning and Expression of Mycobacterium bovis Secreted Protein MPB83 in Escherichia coli [2006]
Jiang, Xiu Yun (Jilin Agricultural University, Changchun, China); Wang, Chun Feng (Jilin Agricultural University, Changchun, China); Wang, Chun Fang (Jilin Agricultural University, Changchun, China); Zhang, Peng Ju (Jilin Agricultural University, Changchun, China); et al.
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The gene encoding MPB83 from Mycobacterium bovis Vallee111 chromosomal DNA was amplified by using polymerase chain reaction (PCR) technique, and the PCR product was approximately 600bp DNA segment. Using TA cloning technique, the PCR product was cloned into pGEM-T vector and the cloning plasmid pGEM-T-83 was constructed successfully. pGEM-T-83 and pET28a(+) were digested by BamHI and EcoRI double enzymes. The purified MPB83 gene was subcloned into the expression vector pET28a(+), and the prokaryotic expression vector pET28a-83 was constructed.
