Cloning and DNA sequence analysis of the glucose transporter gene2 from Iranian Saccharomyces cerevisiae

  Introduction: Saccharomyces cerevisiae has 20 genes that encode hexose transporter proteins including HXT1 to HXT17, GAL2, SNF3 and RGT2. Among these gene families, seven genes (HXT1-HXT7) have important role in alcohol production. The aim of this study was the identification and isolation of HXT2 gene from Saccharomyces cerevisiae genome by PCR technique and cloning into vector containing suitable expression promoter in order to design expression vector as a basis to produce recombinant yeast by transformation.   Materials and Methods: After designing specific oligonucleotides primers, fragment gene amplified by PCR. Gene HXT2 inserted into pTZ57R vector by restriction enzymes EcoRI and HindIII and T4 ligase. After transformation of pTZ57R/THXT2 into E.coli, plasmid recombinant analysis considered. The final further analysis by restriction enzymes digestion and software were evaluated.   Results: HXT2 gene isolated from pTZ57R/THXT2 has correct size in agarose gel electrophoresis. Electrophoresis analysis showed that this gene has correct size on agarose gel. Software study showed that this gene encode proteins with 59.84 KDa molecular weight having 541 amino acids with isoelectric point 8.3.   Conclusion: HXT2 gene by PCR optimization from saccharomyces cerevisiae was isolated and cloned into prokaryotic host. This is the first report of isolation and cloning of this gene by using genetic engineering technique in IRAN that can be used for cloning into suitable expression vector to improve alcohol fermentation yield.