title: Expression and Purification of the Recombinant Cytochrome P450 CYP141 Protein of Mycobacterium Tuberculosis as a Diagnostic Tool and Vaccine Production.
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.
  
  
  OBJECTIVE
  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.
  
  
  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.
authors.affiliation: Department of Microbiology and Immunology, Cellular and Molecular Research Center, Shahrekord University of Medical Sciences, Shahrekord, IR Iran.
authors.forename: Reza
authors.initials: R
authors.lastname: Heidari
authors.affiliation: Department of Microbiology, Shahrekord Branch, Islamic Azad University, Shahrekord, IR Iran.
authors.forename: Mohammad
authors.initials: M
authors.lastname: Rabiee-Faradonbeh
authors.affiliation: Department of Microbiology and Immunology, Cellular and Molecular Research Center, Iran University of Medical Sciences, Tehran, IR Iran.
authors.forename: Davood
authors.initials: D
authors.lastname: Darban-Sarokhalil
authors.affiliation: Department of Microbiology, Kermanshah University of Medical Sciences, Kermanshah, IR Iran.
authors.forename: Amirhooshang
authors.initials: A
authors.lastname: Alvandi
authors.affiliation: Cellular and Molecular Research Center, Shahrekord University of Medical Sciences, Shahrekord, IR Iran.
authors.forename: Narges
authors.initials: N
authors.lastname: Abdian
authors.affiliation: Antimicrobial Resistance Research Center, Department of Medical Microbiology, Mashhad University of Medical Sciences, Mashhad, IR Iran.
authors.forename: Ehsan
authors.initials: E
authors.lastname: Aryan
authors.affiliation: Department of Pathology, Al-Zahra Hospital, Isfahan University of Medical Sciences, Isfahan, IR Iran.
authors.forename: Neda
authors.initials: N
authors.lastname: Soleimani
authors.affiliation: Department of Microbiology and Immunology, Cellular and Molecular Research Center, Shahrekord University of Medical Sciences, Shahrekord, IR Iran.
authors.forename: Abolfazl
authors.initials: A
authors.lastname: Gholipour
keywords: Cytochrome P450 Enzyme System; Gene Expression; Mycobacterium Tuberculosis