title: Construction and Evaluation of an Expression Vector Containing Mtb32C (Rv0125) of Mycobacterium tuberculosis.
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.
authors: Microbiology and Virology Research Center, Avicenna Research Institute & Department of Medical Bacteriology & Virology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.
authors: Maryam Sadat
authors: MS
authors: Nabavinia
authors: Mahboobeh Naderi
authors: MN
authors: Nasab
authors: Zahra
authors: Z
authors: Meshkat
authors: Mohammad
authors: M
authors: Derakhshan
authors: Mehrangiz
authors: M
authors: Khaje-Karamadini
keywords: Cloning vector; Fusion partner; Molecular biotechnology; Mycobacterium tuberculosis; Recombinant protein