کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
2091394 | 1081544 | 2006 | 9 صفحه PDF | دانلود رایگان |
![عکس صفحه اول مقاله: A novel system of genetic transformation allows multiple integrations of a desired gene in Saccharomyces cerevisiae chromosomes A novel system of genetic transformation allows multiple integrations of a desired gene in Saccharomyces cerevisiae chromosomes](/preview/png/2091394.png)
Increasing industrial competitiveness and productivity demand that recombinant yeast strains, used in many different processes, be constantly adapted and/or genetically improved to suit changing requirements. Among yeasts, Saccharomyces cerevisiae is the best-studied organism, and the most frequently employed yeast in industrial processes. In the present study, laboratory strains and industrial S. cerevisiae strains were stably transformed with a novel vector containing the glucoamylase cDNA of Aspergillus awamori flanked by δ-sequences (δGlucoδ), and lacking a positive selection marker. Co-transformation with known plasmids allowed selection by auxotrophic complementation of the leu2 mutation and/or geneticin resistance (G418R). In all cases, several copies of the δGlucoδ vector were inserted into the genome of the yeast cell without selective pressure, showing 100% stability after 80 generations. Transformation frequency of the new vector was similar for S. cerevisiae laboratory strains and industrial wild-type S. cerevisiae strains. This novel genetic transformation system is versatile and suitable to introduce several stable copies of a desired expression cassette into the genome of different S. cerevisiae yeast strains.
Journal: Journal of Microbiological Methods - Volume 67, Issue 3, December 2006, Pages 437–445