Construction of Mtb72F Plasmid as a DNA Vaccine Candidate for Mycobacterium tuberculosis

Authors

  • Aida Gholoobi Department of Modern Sciences and Technologies, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.
  • Mahboubeh Naderinasab Antimicrobial Resistance Research Center, Mashhad University of Medical Sciences, Mashhad, Iran
  • Maryam Sadat Nabavinia Department of Pharmaceutical Biotechnology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran
  • Mohammad Ramezani Nanotechnology Research Center, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran
  • Zahra Meshkat Antimicrobial Resistance Research Center, Mashhad University of Medical Sciences, Mashhad, Iran
Abstract:

Background:  With one-third of the world’s population infected, tuberculosis (TB) is one of the most common infectious diseases and a major public health problem, especially in developing countries. The efficacy of the BCG vaccine for controlling the disease in adults is poor. The development of an effective TB vaccine is a global objective. An effective tuberculosis vaccine should stimulate cellular immunity. DNA vaccines are a new generation of vaccines with the potential to achieve this goal. The aim of this study was to produce a DNA vaccine of Mtb72F.   Methods: mtb32C, mtb39, and mtb32N were cloned into pcDNA3.1 using restriction enzyme digestion and T4 DNA ligase. Colony-PCR and restriction enzyme digestion were performed to detect transformed bacteria. DNA sequencing confirmed the desired gene insertion into the vector. A Chinese hamster ovary (CHO) cell line was transfected with the recombinant plasmid and RT-PCR was performed to assess gene expression. Results: Gel electrophoresis showed the expected amplified gene fragments of 429, 614, and 1200 base pairs (bps) for mtb32C, mtb32N, and mtb39, respectively. Enzyme digestion and gel electrophoresis showed the expected fragments, indicating the desired gene position and orientation in the recombinant plasmid. This finding was verified by DNA sequencing, and RT-PCR demonstrated gene expression in the CHO cell line. Conclusions: An Mtb72F DNA plasmid was successfully constructed. This plasmid may be a candidate for animal immunizations.

Upgrade to premium to download articles

Sign up to access the full text

Already have an account?login

similar resources

Designing and Construction of a Cloning Vector Encoding mtb32C and mpt51 Fragments of Mycobacterium Tuberculosis as a DNA Vaccine Candidate

Background & objective:  Tuberculosis (TB) remains a major cause of death around the world. Bacillus Calmette Guérin (BCG) is the only vaccine used in TB prevention that has a protective effect in children, but its effectiveness declines in adults. Design and development of new vaccines is the most effective way against TB. The aim of this study was to design and construc...

full text

Construction of a Eukaryotic Plasmid Encoding Bacillus anthracis Protective Antigen, a Candidate for DNA Vaccine

Background: DNA immunization with plasmid DNA encoding bacterial, viral, parasitic and tumor antigens has been reported to trigger protective immunity. Objective: To evaluate the use of a DNA immunization strategy for protection against anthrax, a plasmid was constructed. Methods: The partialsequence of protective antigen of Bacillus anthracis, amino acids 175-764, as a potent immunogenic targe...

full text

Construction of a Novel DNA Vaccine Candidate Encoding an HspX-PPE44-EsxV Fusion Antigen of Mycobacterium tuberculosis

Background: Mycobacterium tuberculosis is the causative agent of tuberculosis (TB). Bacille Calmette-Guerin (BCG) vaccine, is not effective in adults, therefore, many efforts have been made to produce an effective adult TB vaccine. The aim of this study was to develop a new tuberculosis DNA vaccine candidate encoding a recombinant HspX-PPE44-EsxV fusion antigen of M. tuberculosis. Methods: ...

full text

Designing and construction of a DNA vaccine encoding tb10.4 gene of Mycobacterium tuberculosis

Background: Tuberculosis (TB) remains as a major cause of death around the world. Construction of a new vaccine against tuberculosis is an effective way to control it. Several vaccines against this disease have been developed. The aim of the present study was to cloning of tb10.4 gene in pcDNA3.1+ plasmid and evaluation of its expression in eukaryotic cells. ...

full text

Cloning, Expression, and Refolding of PPE17 Protein of Mycobacterium Tuberculosis as a Promising Vaccine Candidate

Background: Tuberculosis as a global health problem requires special attention in the contexts of prevention and control. Subunit vaccines are promising strategies to protect burdens of tuberculosis via increasing the BCG protection. The present study aimed to design a vaccine study by means of high-throughput expression and correct refolding of recombinant protein PPE17. Methods: We aimed to c...

full text

Construction of an Expression Plasmid (Vector) Encoding Brucella melitensis Outer Membrane Protein, a Candidate for DNA Vaccine

Background: DNA vaccination with plasmid encoding bacterial, viral, and parasitic immunogens has been shown to be an attractive method to induce efficient immune responses. Bacteria of the genus Brucella are facultative intracellular pathogens for which new and efficient vaccines are needed. Methods: To evaluate the use of a DNA immunization strategy for protection against brucellosis, a pla...

full text

My Resources

Save resource for easier access later

Save to my library Already added to my library

{@ msg_add @}


Journal title

volume 6  issue 1

pages  95- 101

publication date 2017-10

By following a journal you will be notified via email when a new issue of this journal is published.

Keywords

Hosted on Doprax cloud platform doprax.com

copyright © 2015-2023