|Year : 2016 | Volume
| Issue : 5 | Page : 249
Cloning, expression, and purification of recombinant protein MPT-64 from a virulent strain of Mycobacterium bovis in a prokaryotic system
Maryam Mohammadi Tashakkori1, Majid Tebianian2, Mohammad Tabatabaei1, Nader Mosavari1
1 Department of Pathobiology, School of Veterinary Medicine, Shiraz University, Shiraz, Iran
2 Department of Biotechnology, Razi Vaccine and Serum Research Institute, Karaj, Iran
|Date of Web Publication||17-Feb-2017|
PhD of Immunology, Razi Vaccine & Serum Research Institute, P.O.BOX: 31975/148, Karaj
Source of Support: None, Conflict of Interest: None
Objective: Tuberculosis (TB) is a zoonotic infectious disease common to humans and animals that is caused by the rod-shaped acid-fast bacterium Mycobacterium bovis. Rapid and sensitive detection of TB is promoted by specific antigens. Virulent strains of the TB complex from M. bovis contain 16 regions of difference (RD) in their genome that encode important proteins, including major protein of Mycobacterium tuberculosis 64 (MBT-64, which is a primary immune-stimulating antigen encoded by RD-2. In this study, we cloned, expressed, and purified MPT-64 as a potent M. bovis antigen in a prokaryotic system for use in future diagnostic studies.
Methods: The antigenic region of the Mpt64 gene was investigated by bioinformatics methods, cloned into the PQE-30 plasmid, and expressed in Escherichia coli M15 cells, followed by isopropyl β-d-1-thiogalactopyranoside induction. The expressed protein was analyzed sodium dodecyl sulfate polyacrylamide gel electrophoresis and purified using a nickel-affinity column. Biological activity was confirmed by western blot using specific antibodies.
Results: Our data verified the successful cloning of the Mpt64 gene (687-bp segment) via the expression vector and purification of recombinant MPT-64 as a 24-kDa protein.
Conclusion: These results indicated successful expression and purification of recombinant MPT-64 protein in a prokaryotic system. This protein can be used for serological diagnosis, improved detection of pathogenicity and non-pathogenicity between infected cattle, and for verification of suspected cases of bovine TB.
Keywords: Cloning, MPT-64 protein, Mycobacterium bovis, Recombinant protein
|How to cite this article:|
Tashakkori MM, Tebianian M, Tabatabaei M, Mosavari N. Cloning, expression, and purification of recombinant protein MPT-64 from a virulent strain of Mycobacterium bovis in a prokaryotic system. Int J Mycobacteriol 2016;5, Suppl S1:249
|How to cite this URL:|
Tashakkori MM, Tebianian M, Tabatabaei M, Mosavari N. Cloning, expression, and purification of recombinant protein MPT-64 from a virulent strain of Mycobacterium bovis in a prokaryotic system. Int J Mycobacteriol [serial online] 2016 [cited 2020 Mar 28];5, Suppl S1:249. Available from: http://www.ijmyco.org/text.asp?2016/5/5/249/200458
| Conflicts of interest|| |
The authors have no conflicts of interest to declare.