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. 1993 May;13(5):2697–2705. doi: 10.1128/mcb.13.5.2697

Transformation of Saccharomyces cerevisiae with nonhomologous DNA: illegitimate integration of transforming DNA into yeast chromosomes and in vivo ligation of transforming DNA to mitochondrial DNA sequences.

R H Schiestl 1, M Dominska 1, T D Petes 1
PMCID: PMC359643  PMID: 8386316

Abstract

When the yeast Saccharomyces cerevisiae was transformed with DNA that shares no homology to the genome, three classes of transformants were obtained. In the most common class, the DNA was inserted as the result of a reaction that appears to require base pairing between the target sequence and the terminal few base pairs of the transforming DNA fragment. In the second class, no such homology was detected, and the transforming DNA was integrated next to a CTT or GTT in the target; it is likely that these integration events were mediated by topoisomerase I. The final class involved the in vivo ligation of transforming DNA with nucleus-localized linear fragments of mitochondrial DNA.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aguilera A., Klein H. L. HPR1, a novel yeast gene that prevents intrachromosomal excision recombination, shows carboxy-terminal homology to the Saccharomyces cerevisiae TOP1 gene. Mol Cell Biol. 1990 Apr;10(4):1439–1451. doi: 10.1128/mcb.10.4.1439. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Asch D. K., Frederick G., Kinsey J. A., Perkins D. D. Analysis of junction sequences resulting from integration at nonhomologous loci in Neurospora crassa. Genetics. 1992 Apr;130(4):737–748. doi: 10.1093/genetics/130.4.737. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Been M. D., Burgess R. R., Champoux J. J. Nucleotide sequence preference at rat liver and wheat germ type 1 DNA topoisomerase breakage sites in duplex SV40 DNA. Nucleic Acids Res. 1984 Apr 11;12(7):3097–3114. doi: 10.1093/nar/12.7.3097. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Binninger D. M., Skrzynia C., Pukkila P. J., Casselton L. A. DNA-mediated transformation of the basidiomycete Coprinus cinereus. EMBO J. 1987 Apr;6(4):835–840. doi: 10.1002/j.1460-2075.1987.tb04828.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Blanc H. Two modules from the hypersuppressive rho- mitochondrial DNA are required for plasmid replication in yeast. Gene. 1984 Oct;30(1-3):47–61. doi: 10.1016/0378-1119(84)90104-5. [DOI] [PubMed] [Google Scholar]
  6. Boeke J. D., LaCroute F., Fink G. R. A positive selection for mutants lacking orotidine-5'-phosphate decarboxylase activity in yeast: 5-fluoro-orotic acid resistance. Mol Gen Genet. 1984;197(2):345–346. doi: 10.1007/BF00330984. [DOI] [PubMed] [Google Scholar]
  7. Bullock P., Champoux J. J., Botchan M. Association of crossover points with topoisomerase I cleavage sites: a model for nonhomologous recombination. Science. 1985 Nov 22;230(4728):954–958. doi: 10.1126/science.2997924. [DOI] [PubMed] [Google Scholar]
  8. Case M. E. Genetical and molecular analyses of qa-2 transformants in Neurospora crassa. Genetics. 1986 Jul;113(3):569–587. doi: 10.1093/genetics/113.3.569. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Delouya D., Nobrega F. G. Mapping of the ARS-like activity and transcription initiation sites in the non-canonical yeast mitochondrial ori 6 region. Yeast. 1991 Jan;7(1):51–60. doi: 10.1002/yea.320070106. [DOI] [PubMed] [Google Scholar]
  10. Denis C. L., Young E. T. Isolation and characterization of the positive regulatory gene ADR1 from Saccharomyces cerevisiae. Mol Cell Biol. 1983 Mar;3(3):360–370. doi: 10.1128/mcb.3.3.360. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Farrelly F., Butow R. A. Rearranged mitochondrial genes in the yeast nuclear genome. Nature. 1983 Jan 27;301(5898):296–301. doi: 10.1038/301296a0. [DOI] [PubMed] [Google Scholar]
  12. Gahlmann R., Doerfler W. Integration of viral DNA into the genome of the adenovirus type 2-transformed hamster cell line HE5 without loss or alteration of cellular nucleotides. Nucleic Acids Res. 1983 Nov 11;11(21):7347–7361. doi: 10.1093/nar/11.21.7347. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gietz R. D., Sugino A. New yeast-Escherichia coli shuttle vectors constructed with in vitro mutagenized yeast genes lacking six-base pair restriction sites. Gene. 1988 Dec 30;74(2):527–534. doi: 10.1016/0378-1119(88)90185-0. [DOI] [PubMed] [Google Scholar]
  14. Hinnen A., Hicks J. B., Fink G. R. Transformation of yeast. Proc Natl Acad Sci U S A. 1978 Apr;75(4):1929–1933. doi: 10.1073/pnas.75.4.1929. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Holmes D. S., Quigley M. A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem. 1981 Jun;114(1):193–197. doi: 10.1016/0003-2697(81)90473-5. [DOI] [PubMed] [Google Scholar]
  16. Hyman B. C., Cramer J. H., Rownd R. H. Properties of a Saccharomyces cerevisiae mtDNA segment conferring high-frequency yeast transformation. Proc Natl Acad Sci U S A. 1982 Mar;79(5):1578–1582. doi: 10.1073/pnas.79.5.1578. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kraft R., Tardiff J., Krauter K. S., Leinwand L. A. Using mini-prep plasmid DNA for sequencing double stranded templates with Sequenase. Biotechniques. 1988 Jun;6(6):544-6, 549. [PubMed] [Google Scholar]
  18. Kuspa A., Loomis W. F. Tagging developmental genes in Dictyostelium by restriction enzyme-mediated integration of plasmid DNA. Proc Natl Acad Sci U S A. 1992 Sep 15;89(18):8803–8807. doi: 10.1073/pnas.89.18.8803. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Louis E. J., Haber J. E. Evolutionarily recent transfer of a group I mitochondrial intron to telomere regions in Saccharomyces cerevisiae. Curr Genet. 1991 Nov;20(5):411–415. doi: 10.1007/BF00317070. [DOI] [PubMed] [Google Scholar]
  20. Murnane J. P., Yezzi M. J., Young B. R. Recombination events during integration of transfected DNA into normal human cells. Nucleic Acids Res. 1990 May 11;18(9):2733–2738. doi: 10.1093/nar/18.9.2733. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Porter S. E., Champoux J. J. Mapping in vivo topoisomerase I sites on simian virus 40 DNA: asymmetric distribution of sites on replicating molecules. Mol Cell Biol. 1989 Feb;9(2):541–550. doi: 10.1128/mcb.9.2.541. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Qin S. L., Xie A. G., Bonato M. C., McLaughlin C. S. Sequence analysis of the translational elongation factor 3 from Saccharomyces cerevisiae. J Biol Chem. 1990 Feb 5;265(4):1903–1912. [PubMed] [Google Scholar]
  23. Schiestl R. H., Petes T. D. Integration of DNA fragments by illegitimate recombination in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1991 Sep 1;88(17):7585–7589. doi: 10.1073/pnas.88.17.7585. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Skelly P. J., Clark-Walker G. D. Conversion at large intergenic regions of mitochondrial DNA in Saccharomyces cerevisiae. Mol Cell Biol. 1990 Apr;10(4):1530–1537. doi: 10.1128/mcb.10.4.1530. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Szostak J. W., Orr-Weaver T. L., Rothstein R. J., Stahl F. W. The double-strand-break repair model for recombination. Cell. 1983 May;33(1):25–35. doi: 10.1016/0092-8674(83)90331-8. [DOI] [PubMed] [Google Scholar]
  26. Szostak J. W., Wu R. Unequal crossing over in the ribosomal DNA of Saccharomyces cerevisiae. Nature. 1980 Apr 3;284(5755):426–430. doi: 10.1038/284426a0. [DOI] [PubMed] [Google Scholar]
  27. Thorsness P. E., Fox T. D. Escape of DNA from mitochondria to the nucleus in Saccharomyces cerevisiae. Nature. 1990 Jul 26;346(6282):376–379. doi: 10.1038/346376a0. [DOI] [PubMed] [Google Scholar]
  28. Wallis J. W., Chrebet G., Brodsky G., Rolfe M., Rothstein R. A hyper-recombination mutation in S. cerevisiae identifies a novel eukaryotic topoisomerase. Cell. 1989 Jul 28;58(2):409–419. doi: 10.1016/0092-8674(89)90855-6. [DOI] [PubMed] [Google Scholar]
  29. Wilkie T. M., Palmiter R. D. Analysis of the integrant in MyK-103 transgenic mice in which males fail to transmit the integrant. Mol Cell Biol. 1987 May;7(5):1646–1655. doi: 10.1128/mcb.7.5.1646. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. de Zamaroczy M., Bernardi G. The primary structure of the mitochondrial genome of Saccharomyces cerevisiae--a review. Gene. 1986;47(2-3):155–177. doi: 10.1016/0378-1119(86)90060-0. [DOI] [PubMed] [Google Scholar]

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