Cloning and expression of ESAT-6 Gene of Mycobacterium bovis in Escherichia coli

摘要

The gene encoding ESAT-6 from Mycobacterium bovis Vallee111 chromosomal DNA was amplified by using polymerase chain reaction (PCR), the PCR product was approximately 288bp DNA segment. The PCR product was cloned into pGEM-T vector and the cloning plasmid pGEM-T-ESAT-6 was constructed successfully. The purified ESAT-6 gene was subcloned into the expression vector pGEX-4T-3, and the prokaryotic expression Plasmid pGEX-4T-3-ESAT-6 was constructed. Plasmid containing pGEX-4T-3-ESAT-6 was transformed into competence Escherichia coli BL21 (DE3). The bacterium was induced by isopropyl-β-D- thiogalactopyranoside (IPTG) and its lysates