| Autor: |
Guerra OG; Department of Microbiology, Biomedical Sciences Institute, University of São Paulo-USP, Av. Prof. Lineu Prestes 1374, Cidade Universitária, São Paulo, Brazil., Rubio IG, da Silva Filho CG, Bertoni RA, Dos Santos Govea RC, Vicente EJ |
| Jazyk: |
angličtina |
| Zdroj: |
Journal of microbiological methods [J Microbiol Methods] 2006 Dec; Vol. 67 (3), pp. 437-45. Date of Electronic Publication: 2006 Jul 10. |
| DOI: |
10.1016/j.mimet.2006.04.014 |
| Abstrakt: |
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 delta-sequences (deltaGlucodelta), and lacking a positive selection marker. Co-transformation with known plasmids allowed selection by auxotrophic complementation of the leu2 mutation and/or geneticin resistance (G418). In all cases, several copies of the deltaGlucodelta 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. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
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