Properties of Yeast Transformation

作者：James B Hicks, A. Hinnen, Gerald R. Fink · 发表于：Cold Spring Harbor Symposia on Quantitative Biology · 年份：1979 · DOI：10.1101/sqb.1979.043.01.149 · 被引用次数：171 · 研究领域：CRISPR and Genetic Engineering、Transgenic Plants and Applications、Viral Infectious Diseases and Gene Expression in Insects

Previous studies showed that the plasmid pYeleu10 could be stably integrated into yeast chromosomes at several locations and, once integrated, the bacterial plasmid sequences (Co1E1) behaved as simple Mendelian elements. In this paper, the authors describe several additional aspects of yeast transformation along with results showing that integrated bacterial plasmid sequences can be retrieved from the yeast genome and reintroduced into bacteria as replicant plasmids; cotransformation of a single cell by two independent plasmids occurs at high frequencies; sequence homology directs the integration of transforming DNA molecules; transformation frequency can be increased dramatically either by converting plasmids from circular plasmids to linear molecules with restriction endonucleases or by creating new recombinant plasmids containing fragments of the indigenous yeast plasmid (ScpI) inserted into the original hybrid plasmids.