Precise excision of bacterial transposon Tn 5 in yeast

D. A. Gordenin, M. V. Trofimova, O. N. Shaburova, Y. I. Pavlov, Y. O. Chernoff, Y. V. Chekuolene, Y. Y. Proscyavichus, K. V. Sasnauskas, A. A. Janulaitis

Research output: Contribution to journalArticle

14 Scopus citations

Abstract

We have demonstrated that precise excision of bacterial transposon Tn 5 can occur in the yeast, Saccharomyces cerevisiae. Tn 5 insertions in the yeast gene LYS2 were generated by transposon mutagenesis made in Escherichia coli by means of a λ::Tn 5 vector. Nine insertions of Tn 5 into the structural part of the yeast LYS2 gene situated in a shuttle epsiomal plasmid were selected. All the plasmids with a Tn 5 insertion were used to transform yeast strains carrying a deletion of the entire LYS2 gene or a deletion of the part of LYS2 overlapping the point of insertion. All insertions inactivated the LYS2 gene and were able to revert with low (about 10-8) frequencies to lysine prototrophy. Restriction analysis of revertant plasmids revealed them to be indistinguishable from the original plasmid without Tn 5 insertion. DNA sequencing of the regions containing the points of insertions, made for two revertants, proved that Tn 5 excision was completely precise.

Original languageEnglish (US)
Pages (from-to)388-393
Number of pages6
JournalMGG Molecular & General Genetics
Volume213
Issue number2-3
DOIs
StatePublished - Aug 1 1988

Keywords

  • Tn 5
  • Transposon excision
  • Yeast

ASJC Scopus subject areas

  • Genetics

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    Gordenin, D. A., Trofimova, M. V., Shaburova, O. N., Pavlov, Y. I., Chernoff, Y. O., Chekuolene, Y. V., Proscyavichus, Y. Y., Sasnauskas, K. V., & Janulaitis, A. A. (1988). Precise excision of bacterial transposon Tn 5 in yeast. MGG Molecular & General Genetics, 213(2-3), 388-393. https://doi.org/10.1007/BF00339607