Construction and prokaryotic expression of the fusion gene of MAGE-1 and Mycobacterium tuberculosis HSP70

Objective To construct the fusion gene of human MAGE-1 and Mycobacterium tuberculosis heat shock protein 70. The fusion gene was expressed in E. coli. and the fusion protein was purified. Methods the HSP70 and MAGE-1 genes were amplified by PCR. After sequencing, the two genes were cloned into the expression vector pGEX-4T-1 to construct fusion gene expression plasmid pGEX-MAGE-1-TBHSP70. The DH5α contained the expression plasmid was induced by IPIG and the fusion protein was purified with GSTrap FF column. Results The HSP70 gene and MAGE-1 gene segment were amplified successfully from plasmids, and their sequences were identical with that reported in GenBank. The fusion gene expression plasmid pGEX-MAGE-1-TBHSP70 was successfully constructed. The DH5α contained the plasmid could express a M, 125 000 protein after induced by IPTG and the M_r 125 000 fusion protein was purified by GSTrap FF column. Conclusion In this study, the fusion protein of human MAGE-1 segment and Mycobacterium tuberculosis HSP70 was expressed and purified successfully, which prepared the materials for the research of the HSP70 protein as a adjuvant-free carrier.