بیان همزمان ژنهای hbha وMtb32C در سیستم پروکاریوت

تیمورپور, رقیه and پیری دوگاهه, هادی and ارزنلو, محسن and قلوبی, آیدا and سنکیان, مجتبی and مشکات, زهرا (1396) بیان همزمان ژنهای hbha وMtb32C در سیستم پروکاریوت. jundishapur journal of microbiology ــ 11 (2). pp. 1-6. شاپا 2008-3645


Official URL: http://jjmicrobiol.com/en/articles/14030.html


Co-Expression of hbha and mtb32C Genes from Mycobacterium tuberculosis H37Rv in a Prokaryotic System

English 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.

Item Type:Article
زبان سند : انگلیسی
نویسنده اول :رقیه تیمورپور
نویسنده :هادی پیری دوگاهه
نویسنده :محسن ارزنلو
نویسنده مسئول :زهرا مشکات
Additional Information:Impact Factor (2016) 1.017 Indexed in: ISI, Scopus,Chemical Abstract , Embase, EBSCO, IndexCopernicous, Google Scholar, ISC, SID, Barakatkns , MagIran
کلیدواژه ها (انگلیسی):PCR; Interferon-Gamma Release Test; Heparin-Binding Hemagglutinin Protein; Mycobacterium tuberculosis
Subjects:QW Microbiology and Immunology
Divisions:Faculty of Medicine > Department of Basic Sciences > Department of Microbiology
ID Code:9937
Deposited By: Dr Roghayeh Teimourpour
Deposited On:01 Feb 1397 12:58
Last Modified:06 May 1399 08:58

Repository Staff Only: item control page

Document Downloads

More statistics for this item...