Current Genetics

, Volume 7, Issue 2, pp 93–100 | Cite as

Isolation and characterization of yeast DNA repair genes

II. Isolation of plasmids that complement the mutations rad50-1, rad51-1, rad54-3, and rad55-3
  • I. L. Calderon
  • C. R. Contopoulou
  • R. K. Mortimer


Plasmids that complement the yeast mutations rad50-1, rad51-1, rad54-3 and rad55-3 were obtained by transforming strains that carried a leu2 marker and the particular rad mutation, with YEp13 plasmids containing near random yeast DNA inserts. Integration of these plasmids or of fragments of these plasmids was accomplished. Genetic studies using the integrants established the presence of the genes RAD51, RAD54 and RAD55 in the respective plasmids. However, a BamHI subclone of the rad50-1 complementing plasmid failed to integrate at the RAD50 locus, indicating that no homology exists between this fragment and the RAD50 gene.

A BamHI fragment from the RAD54 plasmid was shown to be internal to the RAD54 gene: its integration within a wild type copy of RAD54 causes the cell to become Rad; its excision is X-ray inducible and restores the Rad+ phenotype. Since cells bearing a disrupted copy of RAD54 are able to survive, we conclude that this gene is not essential.

Key words

Yeast RAD genes Cloning 


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  1. Birnboim HC, Doly J (1979) Nucleic Acids Res 7:1513–1523Google Scholar
  2. Boyer HW, Roulland-Dussoix (1969) J Mol Biol 41:459–472Google Scholar
  3. Brendel M, Haynes RH (1973) Mol Gen Genet 125:197–216Google Scholar
  4. Broach JR, Strathern JN, Hicks JB (1979) Gene 8:121–133Google Scholar
  5. Davis RW, Botstein D, Roth JR (1980) Advanced Bacterial Genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, NYGoogle Scholar
  6. Game JC, Mortimer RK (1974) Mutat Res 24:281–292Google Scholar
  7. Haynes RH, Kunz BA (1981) In: Strathern IN, Jones EW, Broach JR (eds) The Molecular Biology of the Yeast Saccharomyces. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY pp 371–414Google Scholar
  8. Hinnen A, Hicks JB, Fink GR (1978) Proc Natl Acad Sci USA 75:1929–1933Google Scholar
  9. Holmes DS, Quigley M (1981) Anal Biochem 114:193–197Google Scholar
  10. Lauer GD, Roberts TM, Klotz LC (1977) J Mol Biol 114:507–526Google Scholar
  11. Lawrence CW (1982) Adv Genet 21:173–254Google Scholar
  12. Lemontt JF (1980) Genetic and physiological factors affecting repair and mutagenesis in yeast. In: deSerres FJ, Generoso WM, Shelby MD (eds) DNA Repair and Mutagenesis in Eukaryotes. Plenum Press, NY, pp 85–120Google Scholar
  13. Mortimer RK, Schild D (1980) Microbiol Rev 44:519–571Google Scholar
  14. Nasmyth KA, Reed SI (1980) Proc Natl Acad Sci USA 77:2119–2123Google Scholar
  15. Nasmyth KA, Tatchell K (1980) Cell 19:753–764Google Scholar
  16. Orr-Weaver TL, Szostak JW, Rothstein RJ (1981) Proc Natl Acad Sci USA 78:6354–6358Google Scholar
  17. Resnick MA (1969) Photochem Photobiol 9:307–312Google Scholar
  18. Scherer S, Davis RW (1979) Proc Natl Acad Sci USA 76:4951–4955Google Scholar
  19. Schild D, Konforti B, Perez C, Gish W, Mortimer RK (1983) Curr Genet 7:85–92Google Scholar
  20. Sherman F, Fink GR, Lawrence CW (1977) Cold Spring Harbor Laboratory, Cold Spring Harbor, NYGoogle Scholar
  21. Strike T (1978) Characterization of Mutants Sensitive to X-rays. PhD thesis, University of California, DavisGoogle Scholar
  22. Tschumper G, Carbon J (1980) Gene 10:157–166Google Scholar

Copyright information

© Springer-Verlag 1983

Authors and Affiliations

  • I. L. Calderon
    • 1
  • C. R. Contopoulou
    • 1
  • R. K. Mortimer
    • 1
  1. 1.Department of Biophysics and Medical PhysicsUniversity of CaliforniaBerkeleyUSA

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