Author/Authors :
Nabavinia، Maryam sadat نويسنده Microbiology and Virology Research Center, Mashhad University of Medical Sciences, Mashhad, IR Iran , , Naderi nasab، Mahboobeh نويسنده , , Meshkat، Zahra نويسنده , , Derakhshan، Mohammad-Hossein نويسنده , , Khaje-Karamadini، Mehrangiz نويسنده Department of Medical Bacteriology and Virology, Qaem Hospital, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran ,
Abstract :
Expressions of recombinant proteins for different applications are important objectives in molecular biotechnology; however, expression of some recombinant proteins is difficult. Several methods have been designed for expression of these proteins. The aim of this study was to construct a vector containing Mtb32C fragment of Mycobacterium tuberculosis (M.tuberculosis) as a fusion partner in order to improve the expression of fused recombinant proteins. Mtb32C was amplified by polymerase chain reaction (PCR). The amplified fragment was ligated into pET21b+ vector. Colony-PCR, enzyme digestion and DNA sequencing methods were used to confirm the recombinant vector. Colony-PCR showed a 420 bp fragment in size corresponding to the correct size of our fragment. In addition the recombinant plasmids sequencing showed the accuracy of the cloned fragment. For confirming the expression, reverse transcriptase (RT)-PCR analysis was performed showing a 420 bp fragment in agarose gel electrophoresis using specific primers. The construction of a vector containing Mtb32C fragment is promising as a fusion partner for future studies as it affected the expression of the fused proteins and increased immune responses against the partner.
