Isolation, cloning and analysis of the glucose transporter gene 7 from Iranian strain of Saccharomyces cerevisiae

authors:

avatar Solmaz Azizi , avatar Alireza Tarinejad , *

Koomesh: Vol.16, issue 3; 356-365
published online: September 28, 2015
article type: Research Article
received: April 02, 2014
accepted: November 01, 2014

how to cite: Azizi S, Tarinejad A. Isolation, cloning and analysis of the glucose transporter gene 7 from Iranian strain of Saccharomyces cerevisiae. koomesh. 2015;16(3):e151259. 

Abstract

  Introduction: The most important and specific known action for Saccharomyces cerevisiae yeast is producing ethanol by alcohol fermentation and that is probably due to its high efficiency for absorption and fermentation of hexose sugars. The S. cerevisiae express 20 genes that encode hexose transporter proteins, including hxt1 -hxt17 , gal2, snf3 and rgt2. Among all those gene families, hxt1-hxt7 have important role in alcohol production. It has been shown that increasing the expression hxt1-hxt7 accelerates alcohol fermentation and therefore, ethanol production. The aim of this study was to identify and isolate hxt7 from Saccharomyces cerevisiae genome, using PCR and cloning it into a vector containing suitable expression promoter in order to produce recombinant yeast by transformation.   Materials and Methods : isolation of hxt7 by specific primers was achieved via PCR. The amplified fragments were cloned into pGEM-T vector and transformed into Escherichia coli and finally, the recombinant plasmids were sent to sequencing.   Results : The nucleotide sequence of open reading frame in gene was revealed a 1713 bp long with a deduced amino acid of 570 residues. The estimated molecular mass and the predicted isoelectric point of the deduced polypeptide were 62.725 kDa and 7.89 respectively.   Conclusion : The deduced protein sequence showed a high similarity to hxt7 sequences registered in NCBI and with the highest percentage of similarity to Hxt7p S.cerevisiae S288C recorded at NCBI with access code NP010629.3.