title: Construction of a Novel DNA Vaccine Candidate Encoding an HspX-PPE44-EsxV Fusion Antigen of Mycobacterium tuberculosis.
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.
authors.affiliation: Antimicrobial Resistance Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.
authors.forename: Bagher
authors.initials: B
authors.lastname: Moradi
authors.affiliation: Immunology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.
authors.forename: Mojtaba
authors.initials: M
authors.lastname: Sankian
authors.affiliation: Immunology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.
authors.forename: Yousef
authors.initials: Y
authors.lastname: Amini
authors.affiliation: Antimicrobial Resistance Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.
authors.forename: Zahra
authors.initials: Z
authors.lastname: Meshkat
keywords: DNA vaccine; EsX; Hsp; Mycobacterium tuberculosis; PPE44