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G418 Selection and stability of cloned genes integrated at chromosomal δ sequences of Saccharomyces cerevisiae
Author(s) -
Wang Xiaohai,
Wang Zhengjun,
Da Silva Nancy A.
Publication year - 2000
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/(sici)1097-0290(19960105)49:1<45::aid-bit6>3.0.co;2-t
Subject(s) - saccharomyces cerevisiae , genetics , biology , gene , yeast , transformation (genetics) , genome , plasmid , selection (genetic algorithm) , artificial intelligence , computer science
The chromosomal δ sequences of the yeast Saccharomyces cerevisiae were employed as recombination sites to integrate the bacterial neo r gene and the yeast SUC2 gene into the yeast genome. A dominate selection method employing the aminoglycoside antibiotic G418 was used. Transformation efficiencies and growth behaviors of the transformants were studied. Transformants were obtained with more than 40 integrations; the majority of insertions were tandem with a maximum of three different insertion sites utilized at one time. After 70–100 generations of growth in nonselective medium, the high copy number SUC2 ‐ neo r integrants were found to be unstable; only minor instability was observed for the neo r and low copy number SUC2 ‐ neo r integrants. © 1996 John Wiley & Sons, Inc.