Abstract

Majority of the shuttle vectors developed for the metabolic engineering of yeast have low copy number, hence the need to be rescued and optimized in Escherichia coli prior to genetic manipulation studies. Methods that are currently available are complicated, contain lot of detergents, time consuming and often yield poor quality plasmid DNA. The present method describes a simple and rapid protocol for isolation of plasmids from yeast using buffer without any detergents. The plasmids isolated by this protocol can be transformed efficiently in E. coli. Further, we also demonstrate that Saccharomyces cerevisiae DNA preparations are best suited for PCR amplification.

Key words: Saccharomyces cerevisiae, plasmid, chromosomal DNA, plasmid rescue, PCR, transformation.