Co-expression of hbha and mtb32C genes from mycobacterium tuberculosis H37Rv in a prokaryotic system

Teimourpour, R. and Peeridogaheh, H. and Arzanlou, M. and Gholoobi, A. and Sankian, M. and Meshkat, Z. (2018) Co-expression of hbha and mtb32C genes from mycobacterium tuberculosis H37Rv in a prokaryotic system. Jundishapur Journal of Microbiology, 11 (2).

[img] Text
Co-expression of hbha and mtb32C genes from mycobacterium tuberculosis H37Rv in a prokaryotic system.pdf

Download (770kB)

Abstract

Background: Heparin-binding hemagglutinin (HBHA) protein is a surface adhesin that mediates the attachment of Mycobacterium tuberculosis to host cells by its own unique, carboxyl-terminal region. The methylated HBHA has a specific motif with a lysine-, alanine-, and proline-rich domain. More recently, it has been shown that HBHA protein has potential activity in stimulating immune responses, and is a promising new candidate for diagnostic applications besides a protective antigen against tuberculosis. Objectives: The aim of this study was to isolate a mycobacterial latency gene (hbha), and subsequently produce its protein as a new antigen for the Interferon-Gamma Release Assay test (IGRAs). Methods: In the present work, hbha and mtb32C genes were isolated from the Mycobacterium tuberculosis H37Rv genome using the polymerase chain reaction (PCR) method. The PCR products and pet21+ vector were digested with specific restriction enzymes and then submitted to the ligation procedure. Escherichia coli BL21-CodonPlus (DE3) competent cells were transformed with the recombinant mtb32C-hbha -pet21+ vector. Expression of recombinant protein (Mtb32C-HBHA) was confirmed with Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) and western blot methods. Results: Detection of a 500-bp gene and sequencing of recombinant pet-mtb32C-hbha vector, all confirmed the accuracy of the cloning procedure. A 36-KDa band of Mtb32C-HBHA protein was also detected by western blotting. Conclusions: In this study, expression of Mtb32C-HBHA protein was successfully done, in the prokaryotic system. Further studies are needed to evaluate the efficacy of recombinant Mtb32C-HBHA protein in diagnosis of latent tuberculosis. © 2018, Jundishapur Journal of Microbiology.

Item Type: Article
Additional Information: Export Date: 16 February 2020 Correspondence Address: Meshkat, Z.; Antimicrobial Resistance Research Center, Mashhad University of Medical SciencesIran; email: meshkatz@mums.ac.ir
Uncontrolled Keywords: Heparin-binding hemagglutinin protein Interferon-gamma release test Mycobacterium tuberculosis PCR Article bacterial cell bacterial gene controlled study gene expression gene identification gene isolation genetic analysis hbha gene interferon gamma release assay latent tuberculosis molecular cloning mtb32C gene nonhuman polyacrylamide gel electrophoresis polymerase chain reaction prokaryote sequence analysis Western blotting
Subjects: WF Respiratory System
QW Microbiology and Immunology
QZ pathology-neoplasms-Genetics
Divisions: Mashhad University of Medical Sciences
Depositing User: lib2 lib2 lib2
Date Deposited: 27 Apr 2020 08:55
Last Modified: 27 Apr 2020 08:55
URI: http://eprints.mums.ac.ir/id/eprint/17560

Actions (login required)

View Item View Item