Expression and purification of the recombinant cytochrome P450 CYP141 protein of mycobacterium tuberculosis as a diagnostic tool and vaccine production

Heidari, R. and Rabiee-Faradonbeh, M. and Darban-Sarokhalil, D. and Alvandi, A. and Abdian, N. and Aryan, E. and Soleimani, N. and Gholipour, A. (2015) Expression and purification of the recombinant cytochrome P450 CYP141 protein of mycobacterium tuberculosis as a diagnostic tool and vaccine production. Iranian Red Crescent Medical Journal, 17 (6). pp. 1-6.

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Abstract

Background: Tuberculosis (TB) is regarded as a health problem worldwide, particularly in developing countries. Mycobacterium tuberculosis (M. tuberculosis) is the cause of this disease. Approximately two billion people worldwide are infected by M. tuberculosis and annually about two million individuals die in consequence. Forty million people are estimated to die because of M. tuberculosis over the next 25 years if the measures for controlling this infection are not extensively developed. In the vaccination field, Bacillus Calmette-Guérin (BCG) is still the most effective vaccine but it shows no efficacy in adult pulmonary patients. One of the other problems regarding TB is its appropriate diagnosis. Objectives: In this experimental study, the recombinant cytochrome P450 CYP141 protein of M. tuberculosis was expressed and purified to be used as a vaccine candidate and diagnostic purpose in subsequent investigations. Materials and Methods: The optimization of the cytochrome P450 CYP141 protein expression was evaluated in different conditions. Then, this protein was purified with a resin column of nickel-nitrilotriacetic acid and investigated via Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) and Western Blotting. Results: The highest expression of the cytochrome P450 CYP141 protein was obtained by the addition of 1 mM of isopropyl β-D-1-thiogalactopyranoside (IPTG) to the bacterial culture grown to an optical density at 600 nm (OD600) of 0.6, 16 hours after induction. This protein was subsequently purified with a purification of higher than 80. The results of Western Blotting indicated that the purified protein was specifically detected. Conclusions: In this experimental study, for the first time in Iran the expression and purification of this recombinant protein was done successfully. This recombinant protein could be used as a vaccine candidate and diagnostic purpose in subsequent investigations. © 2015, Iranian Red Crescent Medical Journal.

Item Type: Article
Additional Information: Cited By :2 Export Date: 16 February 2020 Correspondence Address: Gholipour, A.; Department of Microbiology and Immunology, Cellular and Molecular Research Center, Shahrekord University of Medical SciencesIran; email: gholipourabolfazl@yahoo.com
Uncontrolled Keywords: Cytochrome P450 enzyme system Gene expression Mycobacterium tuberculosis cytochrome P450 cytochrome P450 cyp141 protein isopropyl thiogalactoside nitrilotriacetate nickel recombinant protein unclassified drug Article bacterium culture Escherichia coli experimental study gene sequence genetic transformation human molecular cloning molecular weight optical density polyacrylamide gel electrophoresis protein purification recombinant plasmid vaccine production Western blotting
Subjects: WC Communicable Diseases
QV pharmacology
Divisions: Mashhad University of Medical Sciences
Depositing User: mr lib5 lib5
Date Deposited: 06 May 2020 05:44
Last Modified: 06 May 2020 05:44
URI: http://eprints.mums.ac.ir/id/eprint/18880

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