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. 1984 Jan;4(1):195–198. doi: 10.1128/mcb.4.1.195

Recombinational instability of a chimeric plasmid in Saccharomyces cerevisiae.

M S Whiteway, A Ahmed
PMCID: PMC368675  PMID: 6366510

Abstract

Wild-type strains of Saccharomyces cerevisiae exhibit mitotic recombination between the chimeric plasmid TLC-1 and the endogenous 2mu circle that involves sequence homologies between the two plasmids that are not acted on by the 2mu circle site-specific recombination system. This generalized recombination can be detected because it separates the LEU2 and CAN1 markers of TLC-1 from each other through the formation of a plasmid containing only the S. cerevisiae LEU2 region and the 2mu circle. This derivative plasmid is maintained more stably during vegetative growth than TLC-1, and strains which carry it frequently lose the endogenous 2mu circle. Therefore, TLC-1 can provide a convenient selection for [cir0] cells. Formation of this new plasmid is greatly reduced, but not eliminated, in strains containing the rad52-1 mutation. This indicates that generalized mitotic recombination between plasmid sequences utilizes functions required for chromosomal recombination in S. cerevisiae.

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Selected References

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

  1. Ahmed A., Johansen E. The basis of instability of a revertant of the gal3 insertion of Escherichia coli. J Mol Biol. 1978 May 15;121(2):269–281. doi: 10.1016/s0022-2836(78)80009-6. [DOI] [PubMed] [Google Scholar]
  2. Beggs J. D. Transformation of yeast by a replicating hybrid plasmid. Nature. 1978 Sep 14;275(5676):104–109. doi: 10.1038/275104a0. [DOI] [PubMed] [Google Scholar]
  3. Bolivar F., Rodriguez R. L., Greene P. J., Betlach M. C., Heyneker H. L., Boyer H. W., Crosa J. H., Falkow S. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977;2(2):95–113. [PubMed] [Google Scholar]
  4. Broach J. R., Guarascio V. R., Jayaram M. Recombination within the yeast plasmid 2mu circle is site-specific. Cell. 1982 May;29(1):227–234. doi: 10.1016/0092-8674(82)90107-6. [DOI] [PubMed] [Google Scholar]
  5. Broach J. R., Strathern J. N., Hicks J. B. Transformation in yeast: development of a hybrid cloning vector and isolation of the CAN1 gene. Gene. 1979 Dec;8(1):121–133. doi: 10.1016/0378-1119(79)90012-x. [DOI] [PubMed] [Google Scholar]
  6. Cohen S. N., Chang A. C., Hsu L. Nonchromosomal antibiotic resistance in bacteria: genetic transformation of Escherichia coli by R-factor DNA. Proc Natl Acad Sci U S A. 1972 Aug;69(8):2110–2114. doi: 10.1073/pnas.69.8.2110. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cryer D. R., Eccleshall R., Marmur J. Isolation of yeast DNA. Methods Cell Biol. 1975;12:39–44. doi: 10.1016/s0091-679x(08)60950-4. [DOI] [PubMed] [Google Scholar]
  8. Hartley J. L., Donelson J. E. Nucleotide sequence of the yeast plasmid. Nature. 1980 Aug 28;286(5776):860–865. doi: 10.1038/286860a0. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Jackson J. A., Fink G. R. Gene conversion between duplicated genetic elements in yeast. Nature. 1981 Jul 23;292(5821):306–311. doi: 10.1038/292306a0. [DOI] [PubMed] [Google Scholar]
  11. Jazwinski S. M., Edelman G. M. Replication in vitro of the 2-micrometer DNA plasmid of yeast. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1223–1227. doi: 10.1073/pnas.76.3.1223. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Livingston D. M. Inheritance of the 2 micrometer m DNA plasmid from Saccharomyces. Genetics. 1977 May;86(1):73–84. doi: 10.1093/genetics/86.1.73. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. McDonell M. W., Simon M. N., Studier F. W. Analysis of restriction fragments of T7 DNA and determination of molecular weights by electrophoresis in neutral and alkaline gels. J Mol Biol. 1977 Feb 15;110(1):119–146. doi: 10.1016/s0022-2836(77)80102-2. [DOI] [PubMed] [Google Scholar]
  14. Orr-Weaver T. L., Szostak J. W., Rothstein R. J. Yeast transformation: a model system for the study of recombination. Proc Natl Acad Sci U S A. 1981 Oct;78(10):6354–6358. doi: 10.1073/pnas.78.10.6354. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Prakash S., Prakash L., Burke W., Montelone B. A. Effects of the RAD52 Gene on Recombination in SACCHAROMYCES CEREVISIAE. Genetics. 1980 Jan;94(1):31–50. doi: 10.1093/genetics/94.1.31. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Scherer S., Davis R. W. Recombination of dispersed repeated DNA sequences in yeast. Science. 1980 Sep 19;209(4463):1380–1384. doi: 10.1126/science.6251545. [DOI] [PubMed] [Google Scholar]
  17. Storms R. K., Holowachuck E. W., Friesen J. D. Genetic complementation of the Saccharomyces cerevisiae leu2 gene by the Escherichia coli leuB gene. Mol Cell Biol. 1981 Sep;1(9):836–842. doi: 10.1128/mcb.1.9.836. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Storms R. K., McNeil J. B., Khandekar P. S., An G., Parker J., Friesen J. D. Chimeric plasmids for cloning of deoxyribonucleic acid sequences in Saccharomyces cerevisiae. J Bacteriol. 1979 Oct;140(1):73–82. doi: 10.1128/jb.140.1.73-82.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Struhl K., Stinchcomb D. T., Scherer S., Davis R. W. High-frequency transformation of yeast: autonomous replication of hybrid DNA molecules. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1035–1039. doi: 10.1073/pnas.76.3.1035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Thompson R., Hughes S. G., Broda P. Plasmid identification using specific endonucleases. Mol Gen Genet. 1974;133(2):141–149. doi: 10.1007/BF00264835. [DOI] [PubMed] [Google Scholar]
  21. Von Borstel R. C., Cain K. T., Steinberg C. M. Inheritance of spontaneous mutability in yeast. Genetics. 1971 Sep;69(1):17–27. doi: 10.1093/genetics/69.1.17. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Whelan W. L., Gocke E., Manney T. R. The CAN1 locus of Saccharomyces cerevisiae: fine-structure analysis and forward mutation rates. Genetics. 1979 Jan;91(1):35–51. doi: 10.1093/genetics/91.1.35. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Zakian V. A., Brewer B. J., Fangman W. L. Replication of each copy of the yeast 2 micron DNA plasmid occurs during the S phase. Cell. 1979 Aug;17(4):923–934. doi: 10.1016/0092-8674(79)90332-5. [DOI] [PubMed] [Google Scholar]

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