Work

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

Year: 2013

Type: article

Abstract: 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.

Source: Europe PMC (PubMed Central)

Authors: Shima Mahmoudi, Setareh Mamishi, Mona Ghazi, Reihaneh Hosseinpour Sadeghi, Babak Pourakbari

Institutions: Tehran University of Medical Sciences, Shahid Beheshti University, Shahid Beheshti University of Medical Sciences

Language: English

Topic: Tuberculosis Research and Epidemiology

Subfield: Infectious Diseases

Field: Medicine

Domain: Health Sciences

SDG: Good health and well-being

Open Access status: green

OpenAlex ID: W2161667153

FWCI: 0.906

Cites: 25

Cited by: 17

Related to: 10