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Proc Natl Acad Sci U S A. Mar 1979; 76(3): 1035–1039.
PMCID: PMC383183

High-frequency transformation of yeast: autonomous replication of hybrid DNA molecules.


A set of vector DNAs (Y vectors) useful for the cloning of DNA fragments in Saccharomyces cerevisiae (yeast) and in Escherichia coli are characterized. With these vectors, three modes of yeast transformation are defined. (i) Vectors containing yeast chromosomal DNA sequences (YIp1, YIp5) transform yeast cells at low frequency (1--10 colonies per microgram) and integrate into the genome by homologous recombination; this recombination is reversible. (ii) Hybrids containing endogenous yeast plasmid DNA sequences (YEp2, YEp6) transform yeast cells at much higher frequency (5000--20,000 colonies per microgram). Such molecules replicate autonomously with an average copy number of 5--10 covalently closed circles per yeast cell and also replicate as a chromosomally integrated structure. This DNA may be physically isolated in intact form from either yeast or E. coli and used to transform either organism at high frequency. (iii) Vectors containing a 1.4-kilobase yeast DNA fragment that includes the centromere linked trp1 gene (YRp7) transform yeast with an efficiency of 500--5000 colonies per microgram; such molecules behave as minichromosomes because they replicate autonomously but do not integrate into the genome. The uses of Y vectors for the following genetic manipulations in yeast are discussed: isolation of genes; construction of haploid strains that are merodiploid for a particular DNA sequence; and directed alterations of the yeast genome. General methods for the selection and the analysis of these events are presented.

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Selected References

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  • Struhl K, Cameron JR, Davis RW. Functional genetic expression of eukaryotic DNA in Escherichia coli. Proc Natl Acad Sci U S A. 1976 May;73(5):1471–1475. [PMC free article] [PubMed]
  • Struhl K, Davis RW. Production of a functional eukaryotic enzyme in Escherichia coli: cloning and expression of the yeast structural gene for imidazole-glycerolphosphate dehydratase (his3). Proc Natl Acad Sci U S A. 1977 Dec;74(12):5255–5259. [PMC free article] [PubMed]
  • Ratzkin B, Carbon J. Functional expression of cloned yeast DNA in Escherichia coli. Proc Natl Acad Sci U S A. 1977 Feb;74(2):487–491. [PMC free article] [PubMed]
  • Hinnen A, Hicks JB, Fink GR. Transformation of yeast. Proc Natl Acad Sci U S A. 1978 Apr;75(4):1929–1933. [PMC free article] [PubMed]
  • Cameron JR, Philippsen P, Davis RW. Analysis of chromosomal integration and deletions of yeast plasmids. Nucleic Acids Res. 1977;4(5):1429–1448. [PMC free article] [PubMed]
  • Yanofsky C, Horn V, Bonner M, Stasiowski S. Polarity and enzyme functions in mutants of the first three genes of the tryptophan operon of Escherichia coli. Genetics. 1971 Dec;69(4):409–433. [PMC free article] [PubMed]
  • Murray NE, Brammar WJ, Murray K. Lambdoid phages that simplify the recovery of in vitro recombinants. Mol Gen Genet. 1977 Jan 7;150(1):53–61. [PubMed]
  • Bolivar F, Rodriguez RL, Greene PJ, Betlach MC, Heyneker HL, Boyer HW, Crosa JH, Falkow S. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977;2(2):95–113. [PubMed]
  • Wensink PC, Finnegan DJ, Donelson JE, Hogness DS. A system for mapping DNA sequences in the chromosomes of Drosophila melanogaster. Cell. 1974 Dec;3(4):315–325. [PubMed]
  • Hicks J, Fink GR. Identification of chromosomal location of yeast DNA from hybrid plasmid p Yeleu 10. Nature. 1977 Sep 15;269(5625):265–267. [PubMed]
  • Clark-Walker GD. Isolation of circular DNA from a mitochondrial fraction from yeast. Proc Natl Acad Sci U S A. 1972 Feb;69(2):388–392. [PMC free article] [PubMed]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences


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