Author/Authors :
Teimourpour، Roghayeh نويسنده Antimicrobial Resistance Research Center, Mashhad University of Medical Sciences, Mashhad, IR Iran , , Sadeghian، Ali نويسنده Department of Microbiology and Virology, Antimicrobial Resistance Research Center, Bu Ali Research Center, y of MSchool of Medicine, Mashhad Universitedical Sciences, Mashhad, IR Iran , , Meshkat، Zahra نويسنده , , Esmaelizad، Majid نويسنده Razi Vaccine and serum research institute , , Jabbari، Ahmad Reza نويسنده , , Sankian، Mojtaba نويسنده ,
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.
Objectives: 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.
Materials and 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.
