Stepwise Transformation in Saccharomyces cerevisiae Yeast : Construction of Strains for Transformation and Subsequent Cytoductive Transfer of Plasmid DNA with Mitochondria

概要

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As a general transformation method in Saccharomyces cerevisiae, a "stepwise transformation method" of three steps was developed: conventional transformation of established carrier strains with plasmid DNA, subsequent effective transfer of plasmid to destination hosts either by cytoduction or by meiosis after mating, and finally a simple test for plasmid retention. The several auxotrophic carrier strains constructed were readily transformable, defective in karyogamy (karl), and highly susceptible to cell-wall lytic enzymes. Effective transfer of plasmid DNA from the carrier strains to other strains by cytoduction or meiosis was confirmed, and the target transformants were obtained. The simple method for checking plasrnid retention was based on the cytoductive transfer of plasmid DNA to tester cells which have recessive multiple drug resistance and karl mutations. The stepwise method should largely extend the range of strains which can be transformed and spare time and labor. Its limitations as well as other advantages are also discussed.
日本植物生理学会の論文