جستجوی مقالات مرتبط با کلیدواژه « saccharomyces cerevisiae » در نشریات گروه « بیوتکنولوژی و ژنتیک گیاهی »

The yeast Saccharomyces cerevisiae has 20 genes that encode Hexose Transporter proteins, including HXT1-HXT17, GAL2, SNF3 and RGT2. Two of these genes (SNF3 and RGT2) act as glucose sensors while the HXT1-HXT17 genes function in direct transportation of glucose. Earlier research has shown that alcohol fermentation can be augmented by increasing the expression of these genes, resulting in increasing ethanol production. The aim of this study was the identification and isolation of the Restores Glucose Transport 2 (RGT2) gene from Saccharomyces cerevisiae genome. Specific primers were employed in PCR so as to clone RGT2 into a vector under a suitable expression promoter for recombinant yeast. After gene amplification, ligation was achieved between the amplified fragments and pGEM-T vector and the recombinant colonies were identified by the blue-white screening method. Candidate recombinant plasmids were sequenced. The nucleotide sequence of the open reading frame was found to be 2292 bp long with a deduced amino acid of 763 residues. The estimated molecular mass and the predicted isoelectric point of the deduced polypeptide were 83.173 kDa and 5.68 respectively. The deduced protein sequence showed a high similarity to RGT2 sequences in the NCBI database, especially with P301 strain of Saccharomyces cerevisiae (100 % similarity). Finally, the RGT2 gene was cloned into the pGBKT7 expression vector which is suitable for protein expression in yeast via the restriction sites NcoI and PstI. A phylogenic study of the RGT2 gene and other hexose transporter families showed that this gene has the most similarity with SNF3. Therefore, by isolation, cloning and sequence identification and transformation of this gene into yeast, ethanol production via alcohol fermentation can be increased.

Upon infection of eukaryotic cells, Agrobacterium tumefaciens transfers a piece of its tumor inducing plasmid, T-DNA, to the host cell. The VirD2 virulence protein which is covalently bound to the T-DNA facilitates its transfer, nuclear localization and possibly integration into the host genome in collaboration with the interacting proteins of the host cell. VirD2 is essential for Agrobacterium–mediated transformation of both plants and non-plant cells. Here, using yeast Green Flourescent Protein (yGFP) technology we studied the subcellular localization of VirD2 expressed in yeast Saccharomyces cerevisiae cells. Fluorescence microscopy showed that an N-terminal yGFP fusion of VirD2, is located in the nucleus of yeast. This highlights nuclear localization of VirD2 in yeast and the importance of NLS sequences of C-terminal of VirD2 in this phenomenon.