Document Type: Original Research
Antimicrobial Resistance Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.
Antimicrobial Resistance Research Center, Mashhad University of Medical Sciences, Mashhad, Iran
Background: Tuberculosis (TB) remains as a major cause of death around the world. Construction of a new vaccine against tuberculosis is an effective way to control it. Several vaccines against this disease have been developed. The aim of the present study was to cloning of tb10.4 gene in pcDNA3.1+ plasmid and evaluation of its expression in eukaryotic cells.
Methods: Firstly, tb10.4 fragment was amplified by PCR and the PCR product was digested with restriction enzymes. Next, it was cloned into pcDNA3.1+ plasmid. Following that, pcDNA3.1+/tb10.4 recombinant plasmid was transfected into eukaryotic cells.
Results: 5700 bp band for pcDNA3.1+/tb10.4 recombinant plasmid and 297 bp fragment for tb10.4 were observed. Cloning and transfection were successful and designed recombinant vector was confirmed by sequencing.
Conclusion: Successful cloning provides a basis for the development of new DNA vaccines against tuberculosis. In the current study, the aim was cloning of tb10.4 gene in pcDNA3.1+ plasmid and transfection into eukaryotic cells.