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Updated: Apr 28, 2026
Enrichment of Native and Recombinant Extracellular Vesicles of Mycobacteria
Published on: December 8, 2023
[Construction and identification of a prokaryotic expression vector for Zmp1 gene from Mycobacterium tuberculosis]
Jiming Zhang1, Chun Yang1, Yonglin He1
1Center for Molecular Medicine and Tumor Research, Department of Pathogen Biology, School of Basic Medicine, Chongqing Medical University, Chongqing 400016, China.
Objective:
To construct a prokaryotic expression plasmid for zinc-dependent metalloprotease-1 (Zmp1) gene from Mycobacterium tuberculosis and express the plasmid in E.coli.
Methods:
Zmp1 gene was amplified by PCR using the genome of Bacillus Calmette-Guerin vaccine (BCG) as a template and inserted into a multiple cloning site of prokaryotic expression vector pET32a(+). The constructed prokaryotic expression vector pET32a±Zmp1 was transformed into E.coli BL21(DE3), and the recombinant proteins were expressed via IPTG induction. Finally, the expression of Zmp1 protein was detected by SDS-PAGE and Western blotting.
Results:
Restriction analysis and sequencing proved that the recombinant plasmid pET-32a±Zmp1 was constructed correctly. The relative molecular mass of the expressed recombinant protein was about 94 000 which was the same with that of presumed fusion protein. Recombinant Zmp1 protein showed a specific binding to monoclonal antibody with His tag.
Conclusion:
The prokaryotic expression vector for Zmp1 gene was successfully constructed and the Zmp1 fusion protein was effectively expressed in E. coli BL21(DE3).
