Study on dsRNA Mediated Gene Silencing in Saccharomyces cerevisiae:Suppressing The Expression of GRE3 gene
- VernacularTitle:酿酒酵母中双链RNA介导基因沉默技术研究GRE3基因表达抑制
- Author:
Yan LI
;
Ming YAN
;
Kui WANG
;
Lin XU
;
Wei XU
;
Pingkai OUYANG
- Publication Type:Journal Article
- Keywords:
RNA interference;
Saccharomyces cerevisiae;
GRE3 gene;
aldose reductase
- From:
Progress in Biochemistry and Biophysics
2007;34(3):292-298
- CountryChina
- Language:Chinese
-
Abstract:
RNA silencing is increasingly employed as an experimental strategy to probe for gene function in several organisms. The purpose of the present study was to test the effect of gene silencing by dsRNA in budding yeast Saccharomyces cerevisiae. GRE3 gene encoding an NADPH-dependent aldose reductase was chosen as an example. A recombinant plasmid psiLENT-GRE3 was constructed based on the pESC-LEU backbone and used to transform S. cerevisiae YPH499. The down regulation of GRE3 gene expression by inducing 1 kb RNA duplex and a 136 bp loop was investigated using reverse transcription- PCR. The results showed that double-stranded RNA mediated gene silencing could be used as a functional tool to decrease the expression level of a specific gene in S. cerevisiae, which would contribute to the understanding of RNA interference in budding yeast.
