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

Authors

  • Bagher Moradi Antimicrobial Resistance Research Center, Mashhad University of Medical Sciences, Mashhad, Iran
  • Mojtaba Sankian Immunology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.
  • Yousef Amini Immunology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.
  • Zahra Meshkat Antimicrobial Resistance Research Center, Mashhad University of Medical Sciences, Mashhad, Iran
Abstract:

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: A fusion DNA segment consisting of HspX, linker, PPE44, linker, and EsxV, after codon optimization, was designed. The fusion DNA was cloned and its sequence confirmed. Then, expression of a recombinant pcDNA3.1 (+)/HspX-PPE44-EsxV plasmid in Chinese hamster ovary (CHO) cells was verified by RT-PCR and Western-blot analysis. Results: A 1968 bp band in RT-PCR and a 68 kDa band on Western-blot analysis confirmed transcription and expression of recombinant hspX-ppe44-esxV in eukaryotic cells. Conclusions: A recombinant DNA segment encoding the HspX-PPE44-EsxV fusion antigen of M. tuberculosis was constructed and considered to be tested as a new TB DNA vaccine candidate.

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Journal title

volume 4  issue 2

pages  89- 97

publication date 2016-05

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