title: Construction of a DNA Vaccine Encoding Mtb32C and HBHA Genes of Mycobacterium tuberculosis.
abstract: BACKGROUND
  Tuberculosis (TB) is a contagious disease caused by Mycobacterium tuberculosis. Development of a new vaccine for tuberculosis requires immunogenic antigens capable of inducing suitable and long-lasting T cell immunity. The emergence of multidrugs and extensively drug-resistant strains of M. tuberculosis has made it a global public health concern.
  
  
  OBJECTIVE
  DNA vaccine is a straightforward method to introduce antigens to the host. In the present study, two immunodominant mycobacterial antigens (Mtb32C and HBHA) were isolated and cloned into pcDNA3.1 (+) to design and construct a new DNA vaccine. This vector is capable of producing new potent chimeric protein.
  
  
  METHODS
  Mtb32C (Rv0125) and heparin-binding haemagglutinin adhesion (HBHA) genes were amplified using polymerase chain reaction (PCR) of M. tuberculosis H37Rv genome and ligated into pcDNA3.1 (+). Colony-PCR and restriction enzyme analysis were used to confirm the accuracy of the cloning procedure.
  
  
  RESULTS
  In the current study, recombinant pcDNA3.1 (+) vector containing Mtb32C and HBHA genes was successfully constructed. The desired size of DNA fragment was observed using single and double digestion methods.
  
  
  CONCLUSIONS
  Mtb32C and HBHA genes were successfully isolated from H37Rv genome and cloned into an eukaryotic expression vector. This vector can be considered as a vaccine to evaluate immune responses in animal models.
authors.affiliation: Department of Microbiology and Virology, Antimicrobial Resistance Research Center, Bu Ali Research Center, School of Medicine, Mashhad University of Medical Sciences, Mashhad, IR Iran.
authors.forename: Roghayeh
authors.initials: R
authors.lastname: Teimourpour
authors.affiliation: Department of Microbiology and Virology, Antimicrobial Resistance Research Center, Bu Ali Research Center, School of Medicine, Mashhad University of Medical Sciences, Mashhad, IR Iran.
authors.forename: Ali
authors.initials: A
authors.lastname: Sadeghian
authors.affiliation: Department of Microbiology and Virology, Antimicrobial Resistance Research Center, Bu Ali Research Center, School of Medicine, Mashhad University of Medical Sciences, Mashhad, IR Iran.
authors.forename: Zahra
authors.initials: Z
authors.lastname: Meshkat
authors.affiliation: Genomics and Genetic Engineering Department, Razi Vaccine and Serum Research Institute, Karaj, IR Iran.
authors.forename: Majid
authors.initials: M
authors.lastname: Esmaelizad
authors.affiliation: Immunology Research Center, Mashhad University of Medical Sciences, Mashhad, IR Iran.
authors.forename: Mojtaba
authors.initials: M
authors.lastname: Sankian
authors.affiliation: Genomics and Genetic Engineering Department, Razi Vaccine and Serum Research Institute, Karaj, IR Iran.
authors.forename: Ahmad-Reza
authors.initials: AR
authors.lastname: Jabbari
keywords: HBHA protein; Mtb32 antigen; Mycobacterium tuberculosis; Vaccines, DNA