G418 Selection and stability of cloned genes integrated at chromosomal δ sequences of Saccharomyces cerevisiae

✍ Scribed by Xiaohai Wang; Zhengjun Wang; Nancy A. Da Silva

• Publisher: John Wiley and Sons
• Year: 2000
• Tongue: English
• Weight: 784 KB
• Volume: 49
• Category: Article
• ISSN: 0006-3592
• DOI: 10.1002/(sici)1097-0290(19960105)49:1<45::aid-bit6>3.0.co;2-t

✦ Synopsis

The chromosomal 6 sequences of the yeast Saccharomyces cerevisiae were employed as recombination sites to integrate the bacterial neo' 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 SUM-neo' integrants were found to be unstable; only minor instability was observed for the neo' and low copy number SUC2-neo' integrants.