Journal
JOURNAL OF BIOTECHNOLOGY
Volume 143, Issue 4, Pages 247-254Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jbiotec.2009.08.007
Keywords
Saccharomyces cerevisiae; Vectors; Prototrophic strains; Dominant selectable markers; GFP
Categories
Funding
- Fondazione Telethon-Italy [GP030039, GGP07019]
- MIUR [2006069034-003]
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A set of shuttle yeast vectors containing the dominant selectable markers KanMX4 or HphMX4 cassettes, conferring resistance to geneticin and hygromycin B, respectively, was constructed. Dominant selectable markers are useful for genetic manipulation of natural, wine and industrial strains which do not contain any auxotrophic markers as well as of strains which cannot grow on synthetic mineral medium. Vectors were characterized by (i) copy number, (ii) mitotic stability both in selective and non-selective conditions, (iii) the efficiency and frequency of transformations, (iv) optimal adaptation times in non-selective media, (v) optimum conditions for transformation of various laboratory, commercial and wine strains, and (vi) expression level of an inserted gene. Furthermore we produced GFP-containing vectors that can be used for protein subcellular localization in prototrophic strains. (C) 2009 Elsevier B.V. All rights reserved.
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