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The Yeast Saccharomyces kluyveri as a Recipient Eukaryote in Transkingdom Conjugation: Behavior of Transmitted Plasmids

By In Transconjugants


The prokaryote Escherichia coli successfully conjugated with the eukaryote Saccharomyces kluyveri, which is relatively distant from the species S. cerevisiae. To achieve this transkingdom conjugation, we constructed three types of conjugative plasmids, namely integrative, replicative, and centromere vectors, for S. cerevisiae. By transfer of any of the three plasmids from E. coli, an S. kluyveri Ura- mutant was converted to the Ura+ phenotype. This phenotype was easily lost under nonselective conditions. Southern analysis of the transcon-jugants clearly indicated the presence of the plasmids in many different structures and sizes. The four different types of plasmid vectors for Saccharomy-ces cerevisiae are distinguishable by their mechanisms of rep-lication in the transformant. These are (i) integrative vectors (YIp, autonomously replicating sequence [ARS]-less integra-tive plasmids), (ii) replicating vectors (YRp, ARS-containing plasmids), (iii) centromere vectors (YCp, CEN-containing plasmids), and (iv) episomal vectors (YEp, 2,u ori plasmids). These plasmids frequently do not work in other yeasts because of the species specificity of their chromosomal elements

Year: 1993
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