Kar-mediated plasmid transfer between yeast strains: Alternative to traditional transformation methods

Abstract

Publisher Summary This chapter describes a method, based entirely on yeast mating and replica plating, that allows the transfer of plasmids from one strain to another and eliminates the need for DNA isolation and transformation. Because this method uses a karl mutation defective only in nuclear fusion, as well as markers that select against rare diploids, only haploid progeny are recovered. The method of kar-mediated plasmid transfer is simple, fast, efficient, and does not require any special equipment or elaborate technical manipulations. In addition, it allows the transfer of plasmids into strains that normally do not transform well. Furthermore, depending on the strain background, only minor genetic changes of the strain may be necessary to meet the required marker criteria. Since the donor and recipient strains do not have to be in a common genetic background, this provides the flexibility to transfer plasmids between different laboratory strains. Because of these features, kar-mediated transfer is an excellent solution for plasmid library transfers or for the repeated transfer of the same plasmid into a number of host strains.