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Rep Biochem Mol Biol. 2016 Apr;4(2):89-97.

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

Author information

1
Antimicrobial Resistance Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.
2
Immunology 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.

CONCLUSION:

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.

KEYWORDS:

DNA vaccine; EsX; Hsp; Mycobacterium tuberculosis; PPE44

PMID:
27536702
PMCID:
PMC4986267

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