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. 1983 Oct;3(10):1730–1737. doi: 10.1128/mcb.3.10.1730

Cloning of Saccharomyces cerevisiae DNA replication genes: isolation of the CDC8 gene and two genes that compensate for the cdc8-1 mutation.

C L Kuo, J L Campbell
PMCID: PMC370034  PMID: 6358860

Abstract

The CDC8 gene, whose product is required for DNA replication in Saccharomyces cerevisiae, has been isolated on recombinant plasmids. The yeast vector YCp50 bearing the yeast ARS1, CEN4, and URA3 sequences, to provide for replication, stability, and selection, respectively, was used to prepare a recombinant plasmid pool containing the entire yeast genome. Plasmids capable of complementing the temperature-sensitive cdc8-1 mutation were isolated by transformation of a cdc8-1 mutant and selection for clones able to grow at the nonpermissive temperature. The entire complementing activity is carried on a 0.75-kilobase fragment, as revealed by deletion mapping. This fragment lies 1 kilobase downstream from the well-characterized sup4 gene, a gene known to be genetically linked to CDC8, thus confirming that the cloned gene corresponds to the chromosomal CDC8 gene. Two additional recombinant plasmids that complement the cdc8-1 mutation but that do not contain the 0.75-kilobase fragment or any flanking DNA were also identified in this study. These plasmids may contain genes that compensate for the lack of CDC8 gene product.

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Selected References

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  1. Arendes J., Kim K. C., Sugino A. Yeast 2-microns plasmid DNA replication in vitro: purification of the CDC8 gene product by complementation assay. Proc Natl Acad Sci U S A. 1983 Feb;80(3):673–677. doi: 10.1073/pnas.80.3.673. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Botstein D., Falco S. C., Stewart S. E., Brennan M., Scherer S., Stinchcomb D. T., Struhl K., Davis R. W. Sterile host yeasts (SHY): a eukaryotic system of biological containment for recombinant DNA experiments. Gene. 1979 Dec;8(1):17–24. doi: 10.1016/0378-1119(79)90004-0. [DOI] [PubMed] [Google Scholar]
  3. Cameron J. R., Loh E. Y., Davis R. W. Evidence for transposition of dispersed repetitive DNA families in yeast. Cell. 1979 Apr;16(4):739–751. doi: 10.1016/0092-8674(79)90090-4. [DOI] [PubMed] [Google Scholar]
  4. Chang L. M., Lurie K., Plevani P. A stimulatory factor for yeast DNA polymerase. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 1):587–595. doi: 10.1101/sqb.1979.043.01.065. [DOI] [PubMed] [Google Scholar]
  5. Dagert M., Ehrlich S. D. Prolonged incubation in calcium chloride improves the competence of Escherichia coli cells. Gene. 1979 May;6(1):23–28. doi: 10.1016/0378-1119(79)90082-9. [DOI] [PubMed] [Google Scholar]
  6. Goodman H. M., Olson M. V., Hall B. D. Nucleotide sequence of a mutant eukaryotic gene: the yeast tyrosine-inserting ochre suppressor SUP4-o. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5453–5457. doi: 10.1073/pnas.74.12.5453. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hartwell L. H. Genetic control of the cell division cycle in yeast. II. Genes controlling DNA replication and its initiation. J Mol Biol. 1971 Jul 14;59(1):183–194. doi: 10.1016/0022-2836(71)90420-7. [DOI] [PubMed] [Google Scholar]
  8. Hartwell L. H. Three additional genes required for deoxyribonucleic acid synthesis in Saccharomyces cerevisiae. J Bacteriol. 1973 Sep;115(3):966–974. doi: 10.1128/jb.115.3.966-974.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hsu Y. P., Kohlhaw G. B. Overproduction and control of the LEU2 gene product, beta-isopropylmalate dehydrogenase, in transformed yeast strains. J Biol Chem. 1982 Jan 10;257(1):39–41. [PubMed] [Google Scholar]
  10. Ito H., Fukuda Y., Murata K., Kimura A. Transformation of intact yeast cells treated with alkali cations. J Bacteriol. 1983 Jan;153(1):163–168. doi: 10.1128/jb.153.1.163-168.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kelley W. S., Chalmers K., Murray N. E. Isolation and characterization of a lambdapolA transducing phage. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5632–5636. doi: 10.1073/pnas.74.12.5632. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kobori J. A., Kornberg A. The Escherichia coli dnaC gene product. I. Overlapping of the dnaC proteins of Escherichia coli and Salmonella typhimurium by cloning into a high copy number plasmid. J Biol Chem. 1982 Nov 25;257(22):13757–13762. [PubMed] [Google Scholar]
  13. Kuo C. L., Campbell J. L. Purification of the cdc8 protein of Saccharomyces cerevisiae by complementation in an aphidicolin-sensitive in vitro DNA replication system. Proc Natl Acad Sci U S A. 1982 Jul;79(14):4243–4247. doi: 10.1073/pnas.79.14.4243. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Lawrence C. W., Sherman F., Jackson M., Gilmore R. A. Mapping and gene conversion studies with the structural gene for iso-1-cytochrome C in yeast. Genetics. 1975 Dec;81(4):615–629. doi: 10.1093/genetics/81.4.615. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Livingston D. M., Kupfer D. M. Control of Saccharomyces cerevisiae 2microN DNA replication by cell division cycle genes that control nuclear DNA replication. J Mol Biol. 1977 Oct 25;116(2):249–260. doi: 10.1016/0022-2836(77)90215-7. [DOI] [PubMed] [Google Scholar]
  16. Mortimer R. K., Hawthorne D. C. Genetic Mapping in Saccharomyces IV. Mapping of Temperature-Sensitive Genes and Use of Disomic Strains in Localizing Genes. Genetics. 1973 May;74(1):33–54. doi: 10.1093/genetics/74.1.33. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Nasmyth K. A., Reed S. I. Isolation of genes by complementation in yeast: molecular cloning of a cell-cycle gene. Proc Natl Acad Sci U S A. 1980 Apr;77(4):2119–2123. doi: 10.1073/pnas.77.4.2119. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Newlon C. S., Fangman W. L. Mitochondrial DNA synthesis in cell cycle mutants of Saccharomyces cerevisiae. Cell. 1975 Aug;5(4):423–428. doi: 10.1016/0092-8674(75)90061-6. [DOI] [PubMed] [Google Scholar]
  19. Panasenko S. M., Cameron J. R., Davis R. W., Lehman I. R. Five hundredfold overproduction of DNA ligase after induction of a hybrid lambda lysogen constructed in vitro. Science. 1977 Apr 8;196(4286):188–189. doi: 10.1126/science.322281. [DOI] [PubMed] [Google Scholar]
  20. Scherer S., Davis R. W. Replacement of chromosome segments with altered DNA sequences constructed in vitro. Proc Natl Acad Sci U S A. 1979 Oct;76(10):4951–4955. doi: 10.1073/pnas.76.10.4951. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Shortle D., Haber J. E., Botstein D. Lethal disruption of the yeast actin gene by integrative DNA transformation. Science. 1982 Jul 23;217(4557):371–373. doi: 10.1126/science.7046050. [DOI] [PubMed] [Google Scholar]
  22. Stinchcomb D. T., Mann C., Davis R. W. Centromeric DNA from Saccharomyces cerevisiae. J Mol Biol. 1982 Jun 25;158(2):157–190. doi: 10.1016/0022-2836(82)90427-2. [DOI] [PubMed] [Google Scholar]

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