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

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TEIMOURPOUR ROGHAYEH; peeridogaheh hadi; ARZANLOU MOHSEN; GHOLOOBI AIDA; SANKIAN MOJTABA; MESHKAT ZAHRA

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Journal: Jundishapur Journal of Microbiology Year:2018 | Volume:11 | Issue:2 Page(s): 0-0

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Title

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

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Keywords

PCR

Interferon-Gamma Release Test

Heparin-Binding Hemagglutinin Protein

Mycobacterium tuberculosis

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.

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APA: Copy

TEIMOURPOUR, ROGHAYEH, peeridogaheh, hadi, ARZANLOU, MOHSEN, GHOLOOBI, AIDA, SANKIAN, MOJTABA, & MESHKAT, ZAHRA. (2018). Co-Expression of hbha and mtb32C Genes from Mycobacterium tuberculosis H37Rv in a Prokaryotic System. JUNDISHAPUR JOURNAL OF MICROBIOLOGY (JJM), 11(2), 0-0. SID. https://sid.ir/paper/317948/en

Vancouver: Copy

TEIMOURPOUR ROGHAYEH, peeridogaheh hadi, ARZANLOU MOHSEN, GHOLOOBI AIDA, SANKIAN MOJTABA, MESHKAT ZAHRA. Co-Expression of hbha and mtb32C Genes from Mycobacterium tuberculosis H37Rv in a Prokaryotic System. JUNDISHAPUR JOURNAL OF MICROBIOLOGY (JJM)[Internet]. 2018;11(2):0-0. Available from: https://sid.ir/paper/317948/en

IEEE: Copy

ROGHAYEH TEIMOURPOUR, hadi peeridogaheh, MOHSEN ARZANLOU, AIDA GHOLOOBI, MOJTABA SANKIAN, and ZAHRA MESHKAT, “Co-Expression of hbha and mtb32C Genes from Mycobacterium tuberculosis H37Rv in a Prokaryotic System,” JUNDISHAPUR JOURNAL OF MICROBIOLOGY (JJM), vol. 11, no. 2, pp. 0–0, 2018, [Online]. Available: https://sid.ir/paper/317948/en

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