Illegitimate integration of single-stranded DNA in Saccharomyces cerevisiae |
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Authors: | K Gjuraci and Z Zgaga |
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Institution: | (1) Laboratory of Biology and Microbial Genetics, Faculty of Food Technology and Biotechnology, University of Zagreb, Pierottijeva 6, 10.000 Zagreb, Croatia, |
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Abstract: | We studied illegitimate recombination by transforming yeast with a single-stranded (ss) non-replicative plasmid. Plasmid
pCW12, containing the ARG4gene, was used for transformation of yeast strains deleted for the ARG4, either in native (circular) form or after linearization within the vector sequence by the restriction enzyme ScaI. Both circular and linearized ss plasmids were shown to be much more efficient in illegitimate integration than their double-stranded
(ds) counterparts and more than two-thirds of the transformants analysed contained multiple tandem integrations of the plasmid.
Pulsed-field gel electrophoresis of genomic DNA revealed significant changes in the karyotype of some transformants. Plasmid
DNA was frequently detected on more than one chromosome and on mitotically unstable, autonomously replicating elements. Our
results show that the introduction of nonhomologous ss DNA into yeast cells can lead to different types of alterations in
the yeast genome.
Received: 9 February 1996/Accepted: 7 July 1996 |
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Keywords: | Illegitimate recombination Single-stranded plasmids Saccharomyces cerevisiae |
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