title: [Construction and identification of a prokaryotic expression vector for Zmp1 gene from Mycobacterium tuberculosis].
abstract: 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.
  
  
  CONCLUSIONS
  The prokaryotic expression vector for Zmp1 gene was successfully constructed and the Zmp1 fusion protein was effectively expressed in E. coli BL21(DE3).
authors.affiliation: Center for Molecular Medicine and Tumor Research, Department of Pathogen Biology, School of Basic Medicine, Chongqing Medical University, Chongqing 400016, China.
authors.forename: Jiming
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Center for Molecular Medicine and Tumor Research, Department of Pathogen Biology, School of Basic Medicine, Chongqing Medical University, Chongqing 400016, China.
authors.forename: Chun
authors.initials: C
authors.lastname: Yang
authors.affiliation: Center for Molecular Medicine and Tumor Research, Department of Pathogen Biology, School of Basic Medicine, Chongqing Medical University, Chongqing 400016, China.
authors.forename: Yonglin
authors.initials: Y
authors.lastname: He
authors.affiliation: Center for Molecular Medicine and Tumor Research, Department of Pathogen Biology, School of Basic Medicine, Chongqing Medical University, Chongqing 400016, China.
authors.forename: Liuqing
authors.initials: L
authors.lastname: Mu
authors.affiliation: Center for Molecular Medicine and Tumor Research, Department of Pathogen Biology, School of Basic Medicine, Chongqing Medical University, Chongqing 400016, China.
authors.forename: Chunyan
authors.initials: C
authors.lastname: Zhang
authors.affiliation: Center for Molecular Medicine and Tumor Research, Department of Pathogen Biology, School of Basic Medicine, Chongqing Medical University, Chongqing 400016, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Fan
authors.affiliation: Center for Molecular Medicine and Tumor Research, Department of Pathogen Biology, School of Basic Medicine, Chongqing Medical University, Chongqing 400016, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Xu
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics*; D015153:Blotting, Western; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics*; D045726:Metalloproteases / Q000235:genetics*; D009169:Mycobacterium tuberculosis / Q000235:genetics / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial; D011993:Recombinant Fusion Proteins; D045726:Metalloproteases; C568073:Zmp1 protein, Mycobacterium tuberculosis