𝔖 Bobbio Scriptorium
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Nature of abortive transformation inSaccharomyces cerevisiae

✍ Scribed by Wendy Y. Yap; Robert H. Schiestl

Book ID
104790234
Publisher
Springer-Verlag
Year
1995
Tongue
English
Weight
361 KB
Volume
28
Category
Article
ISSN
0172-8083

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✦ Synopsis

Disruption mutagenesis by homologous recombination in Saccharomyces cerevisiae is carried out by transforming-DNA fragments containing the target gene disrupted by a selectable marker. A large number of transient (abortive) transformants are often formed that may hinder the isolation of integrants containing the gene disruption. We show that abortive transformants result from re-circularization of the linear transforming-DNA in vivo. Their number was greatly reduced when the cut DNA could not readily re-ligate, either by digestions that gave non-compatible or blunt ends, or by de-phosphorylation. In addition, true integrants could be readily distinguished from abortive transformants through replica plating onto selective media. Enhanced disruption-mutagenesis was also observed when non-compatible ends were generated in an ARS-containing insertion vector.

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