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PMID: 2847007 Published · ppublish English Journal Article

Precise excision of bacterial transposon Tn5 in yeast.

Molecular & general genetics : MGG ·Vol. 213 ·No. 2-3 ·1988-08-00 ·Pages 388-93

Gordenin DA, Trofimova MV, Shaburova ON, Pavlov YI, Chernoff YO, Chekuolene YV, Proscyavichus YY, Sasnauskas KV, Janulaitis AA

Abstract

We have demonstrated that precise excision of bacterial transposon Tn5 can occur in the yeast, Saccharomyces cerevisiae. Tn5 insertions in the yeast gene LYS2 were generated by transposon mutagenesis made in Escherichia coli by means of a lambda::Tn5 vector. Nine insertions of Tn5 into the structural part of the yeast LYS2 gene situated in a shuttle episomal plasmid were selected. All the plasmids with a Tn5 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 Tn5 insertion. DNA sequencing of the regions containing the points of insertions, made for two revertants, proved that Tn5 excision was completely precise.

MeSH Terms
DNA Transposable Elements DNA, Bacterial/genetics Escherichia coli/genetics Genes, Bacterial Genes, Fungal Plasmids Saccharomyces cerevisiae/genetics Transformation, Genetic
Chemicals
DNA Transposable Elements DNA, Bacterial
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Gordenin D A
Department of Genetics, State University of Leningrad, USSR.
Trofimova M V
Shaburova O N
Pavlov Y I
Chernoff Y O
Chekuolene Y V
Proscyavichus Y Y
Sasnauskas K V
Janulaitis A A
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Article Info
Journal
Molecular & general genetics : MGG
Abbr.
Mol Gen Genet
ISSN
0026-8925
DOI
10.1007/BF00339607
Published
1988-08-00
Pages
388-93
Language
English
Region
Germany
NLM ID
0125036
Subset
IM
External Links
PubMed source DOI: 10.1007/BF00339607
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