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Jundishapur Journal of Microbiology
Ahvaz Jundishapur University of Medical Sciences
e-ISSN 2008-4161
p-ISSN 2008-3645
Cloning, expression and purification of early secretory antigenic target 6kDa protein (ESAT-6) of Mycobacterium tuberculosis
Authors
Zahra Farshadzadeh1,*, Mojtaba Sankian2, Forough Yousefi2, Aida Gholobi2, Reza Zarif2, Mahboobeh Naderi nasab2, Tahereh Rashed2, Abdolreza Varasteh2
1Infectious and Tropical Diseases Research Center, Ahvaz Jundishapur University of Medical Sciences, Zahra.farshadzadeh@gmail.com, Iran
2Booali Immunology Research Center Mashhad University of Medical Sciences, Iran
*Corresponding Author: Infectious and Tropical Diseases Research Center, Ahvaz Jundishapur University of Medical Sciences, Zahra.farshadzadeh@gmail.com, Iran. Email: Zahra.farshadzadeh@gmail.com
Published online:Apr 30, 2010
Article type:Research Article
Received:Aug 01, 2009
Accepted:Feb 01, 2010
How to Cite:Farshadzadeh Z, Sankian M, Yousefi F, Gholobi A, Zarif R, et al. Cloning, expression and purification of early secretory antigenic target 6kDa protein (ESAT-6) of Mycobacterium tuberculosis. Jundishapur J Microbiol. 2010;3(2):. doi:
Abstract
Introduction and objective: The early secretory antigenic target 6kDa protein (ESAT-6) antigen from Mycobacterium tuberculosis is a dominant target for cell-mediated immunity in the early phase of tuberculosis (TB) in TB patients. ESAT-6 was found to distinguish TB patients from BCG-vaccinated donors. The aim of this study was cloning and expression of ESAT-6 of M. tuberculosis.
Materials and methods: DNA was extracted from M. tuberculosis H37Rv. PCR was performed using gene-specific oligonucleotide primers and the PCR products were inserted into the pET102/D vector and transferred into Escherichia coli strain TOPO10. The recombinant plasmids transferred into E. coli strain BL21.
Results: The transformed plasmid into E. coli strain BL21 was effectively expressed. The expressed fusion protein (23kDa on SDS-PAGE) was found almost entirely in the soluble form and the recombinant protein was purified by Ni-NTA column.
Conclusion: We successfully cloned and expressed ESAT-6 protein of M. tuberculosis in E. coli. As a specific antigen, it can be useful for diagnosis of both active and latent tuberculosis with ELISA in future.
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Copyright
© 2010, Author(s). This open-access article is available under the Creative Commons Attribution 4.0 (CC BY 4.0) International License (https://creativecommons.org/licenses/by/4.0/), which allows for unrestricted use, distribution, and reproduction in any medium, provided that the original work is properly cited.
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