• Users Online: 181
  • Home
  • Print this page
  • Email this page


 
 Table of Contents  
ARTICLE
Year : 2016  |  Volume : 5  |  Issue : 5  |  Page : 249

Cloning, expression, and purification of recombinant protein MPT-64 from a virulent strain of Mycobacterium bovis in a prokaryotic system


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 Publication17-Feb-2017

Correspondence Address:
Majid Tebianian
PhD of Immunology, Razi Vaccine & Serum Research Institute, P.O.BOX: 31975/148, Karaj
Iran
Login to access the Email id

Source of Support: None, Conflict of Interest: None


DOI: 10.1016/j.ijmyco.2016.10.027

Rights and Permissions
  Abstract 


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 2019 Nov 20];5, Suppl S1:249. Available from: http://www.ijmyco.org/text.asp?2016/5/5/249/200458




  Conflicts of interest Top


The authors have no conflicts of interest to declare.






 

Top
 
 
  Search
 
Similar in PUBMED
   Search Pubmed for
   Search in Google Scholar for
 Related articles
Access Statistics
Email Alert *
Add to My List *
* Registration required (free)

 
  In this article
Abstract
Conflicts of int...

 Article Access Statistics
    Viewed412    
    Printed6    
    Emailed0    
    PDF Downloaded44    
    Comments [Add]    

Recommend this journal