Cloning, expression and purification of Mycobacterium tuberculosis ESAT-6 and CFP-10 antigens

BACKGROUND AND OBJECTIVES: ESAT-6 (6-kDaearly secretory antigenic target) and CFP-10 (10-kDa culture filtrate protein) have been described as dominant antigens recognized by T-cells and considered as virulence factors in Mycobacterium tuberculosis. The aim of this study was to clone, express and pur...

Descripción completa

Detalles Bibliográficos
Autores principales: Mahmoudi, Shima, Mamishi, Setareh, Ghazi, Mona, Hosseinpour Sadeghi, Reihaneh, Pourakbari, Babak
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Tehran University of Medical Sciences 2013
Materias:
Medical Sciences
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4385163/
https://www.ncbi.nlm.nih.gov/pubmed/25848507
_version_ 1782365018579271680
author Mahmoudi, Shima
Mamishi, Setareh
Ghazi, Mona
Hosseinpour Sadeghi, Reihaneh
Pourakbari, Babak
author_facet Mahmoudi, Shima
Mamishi, Setareh
Ghazi, Mona
Hosseinpour Sadeghi, Reihaneh
Pourakbari, Babak
author_sort Mahmoudi, Shima
collection PubMed
description BACKGROUND AND OBJECTIVES: ESAT-6 (6-kDaearly secretory antigenic target) and CFP-10 (10-kDa culture filtrate protein) have been described as dominant antigens recognized by T-cells and considered as virulence factors in Mycobacterium tuberculosis. The aim of this study was to clone, express and purify recombinant ESAT-6 andCFP-10 proteins of M. tuberculosis in soluble form. MATERIALS AND METHODS: ESAT-6 andCFP-10 genes were amplified by PCR, cloned into pET32a (+) vector, and overexpress-ed using isopropyl-beta-D-thiogalactopyranoside in E. coli BL21 (DE3). ESAT-6 andCFP-10 proteins were purified by Ni-NTA affinity chromatography and were detected by anti- ESAT-6 and anti -CFP-10 antibodies. RESULTS: ESAT-6 andCFP-10 genes were successfully expressed and purified. Anti- ESAT-6 and anti-CFP-10 antibodies were produced after induction of immunization against purified ESAT-6 andCFP-10 proteins in rabbit. CONCLUSION: In this study, we cloned, expressed and purified sufficient amounts of ESAT-6 andCFP-10 and it would be tested for the development of diagnostic kit for M. tuberculosis in future.
format Online
Article
Text
id pubmed-4385163
institution National Center for Biotechnology Information
language English
publishDate 2013
publisher Tehran University of Medical Sciences
record_format MEDLINE/PubMed
spelling pubmed-43851632015-04-06 Cloning, expression and purification of Mycobacterium tuberculosis ESAT-6 and CFP-10 antigens Mahmoudi, Shima Mamishi, Setareh Ghazi, Mona Hosseinpour Sadeghi, Reihaneh Pourakbari, Babak Iran J Microbiol Medical Sciences BACKGROUND AND OBJECTIVES: ESAT-6 (6-kDaearly secretory antigenic target) and CFP-10 (10-kDa culture filtrate protein) have been described as dominant antigens recognized by T-cells and considered as virulence factors in Mycobacterium tuberculosis. The aim of this study was to clone, express and purify recombinant ESAT-6 andCFP-10 proteins of M. tuberculosis in soluble form. MATERIALS AND METHODS: ESAT-6 andCFP-10 genes were amplified by PCR, cloned into pET32a (+) vector, and overexpress-ed using isopropyl-beta-D-thiogalactopyranoside in E. coli BL21 (DE3). ESAT-6 andCFP-10 proteins were purified by Ni-NTA affinity chromatography and were detected by anti- ESAT-6 and anti -CFP-10 antibodies. RESULTS: ESAT-6 andCFP-10 genes were successfully expressed and purified. Anti- ESAT-6 and anti-CFP-10 antibodies were produced after induction of immunization against purified ESAT-6 andCFP-10 proteins in rabbit. CONCLUSION: In this study, we cloned, expressed and purified sufficient amounts of ESAT-6 andCFP-10 and it would be tested for the development of diagnostic kit for M. tuberculosis in future. Tehran University of Medical Sciences 2013-12 /pmc/articles/PMC4385163/ /pubmed/25848507 Text en Copyright: © Iranian Journal of Microbiology & Tehran University of Medical Sciences This work is licensed under a Creative Commons Attribution-NonCommercial 3.0 Unported License which allows users to read, copy, distribute and make derivative works for non-commercial purposes from the material, as long as the author of the original work is cited properly.
spellingShingle Medical Sciences
Mahmoudi, Shima
Mamishi, Setareh
Ghazi, Mona
Hosseinpour Sadeghi, Reihaneh
Pourakbari, Babak
Cloning, expression and purification of Mycobacterium tuberculosis ESAT-6 and CFP-10 antigens
title Cloning, expression and purification of Mycobacterium tuberculosis ESAT-6 and CFP-10 antigens
title_full Cloning, expression and purification of Mycobacterium tuberculosis ESAT-6 and CFP-10 antigens
title_fullStr Cloning, expression and purification of Mycobacterium tuberculosis ESAT-6 and CFP-10 antigens
title_full_unstemmed Cloning, expression and purification of Mycobacterium tuberculosis ESAT-6 and CFP-10 antigens
title_short Cloning, expression and purification of Mycobacterium tuberculosis ESAT-6 and CFP-10 antigens
title_sort cloning, expression and purification of mycobacterium tuberculosis esat-6 and cfp-10 antigens
topic Medical Sciences
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4385163/
https://www.ncbi.nlm.nih.gov/pubmed/25848507
work_keys_str_mv AT mahmoudishima cloningexpressionandpurificationofmycobacteriumtuberculosisesat6andcfp10antigens
AT mamishisetareh cloningexpressionandpurificationofmycobacteriumtuberculosisesat6andcfp10antigens
AT ghazimona cloningexpressionandpurificationofmycobacteriumtuberculosisesat6andcfp10antigens
AT hosseinpoursadeghireihaneh cloningexpressionandpurificationofmycobacteriumtuberculosisesat6andcfp10antigens
AT pourakbaribabak cloningexpressionandpurificationofmycobacteriumtuberculosisesat6andcfp10antigens

Ejemplares similares