关键词: 分枝杆菌,结核, Rv1009基因, 克隆, 表达

Abstract: Objective To clone Rv1009 gene segment from Mycobacterium tuberculosis and induce its prokaryotic expression. Methods Polymerase chain reaction (PCR) was used to amplify the encoding gene of Rv1009 protein from Mycobacterium tuberculosis H37Rv,and the amplified DNA fragment was inserted into plasmid pGEX-4T-2. The recombinant was transformed into E. coli BL21,and then was induced for the expression of the Rv1009 fusion protein by IPTG. Results The Length of the amplified DNA fragment was 1300bp, sequence analysis showed the amplified fragment was identical With what we designed. The recombinant plasmid was successfully constructed. A 64kD fusion protein was obtained after E. coli BL21 containing recombinant plasmid was induced by 0.3 mmol/L IPTG and accounted for 22.8% of the total protein. Conclusion The colne and expression of Rv1009 were successful, which lays a basis for further study on Mycobacterium tuberculosis.

Key words: Mycobacterium tuberculosis, Rv1009 gene, gene recombination