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PMID: 2659436 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae.

Genetics ·Vol. 122 ·No. 1 ·1989-05-00 ·Pages 19-27

Sikorski RS, Hieter P

Abstract

A series of yeast shuttle vectors and host strains has been created to allow more efficient manipulation of DNA in Saccharomyces cerevisiae. Transplacement vectors were constructed and used to derive yeast strains containing nonreverting his3, trp1, leu2 and ura3 mutations. A set of YCp and YIp vectors (pRS series) was then made based on the backbone of the multipurpose plasmid pBLUESCRIPT. These pRS vectors are all uniform in structure and differ only in the yeast selectable marker gene used (HIS3, TRP1, LEU2 and URA3). They possess all of the attributes of pBLUESCRIPT and several yeast-specific features as well. Using a pRS vector, one can perform most standard DNA manipulations in the same plasmid that is introduced into yeast.

MeSH Terms
Centromere Culture Media DNA, Fungal/genetics Genetic Vectors Plasmids Restriction Mapping Saccharomyces cerevisiae/genetics Transformation, Genetic
Chemicals
Culture Media DNA, Fungal
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Sikorski R S
Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
Hieter P
References (30)
30 references, click to expand
  1. Yeast LEU2. Repression of mRNA levels by leucine and primary structure of the gene product.
    J Biol Chem. 1984 Jul 10;259(13):8059-62 PMID: 6330094
  2. Centromeric DNA from chromosome VI in Saccharomyces cerevisiae strains.
    EMBO J. 1982;1(12):1605-11 PMID: 6327282
  3. Sequences that regulate the divergent GAL1-GAL10 promoter in Saccharomyces cerevisiae.
    Mol Cell Biol. 1984 Aug;4(8):1440-8 PMID: 6092912
  4. A positive selection for mutants lacking orotidine-5'-phosphate decarboxylase activity in yeast: 5-fluoro-orotic acid resistance.
    Mol Gen Genet. 1984;197(2):345-6 PMID: 6394957
  5. Plasmids pEMBLY: new single-stranded shuttle vectors for the recovery and analysis of yeast DNA sequences.
    Gene. 1985;35(1-2):27-32 PMID: 3896935
  6. Functional selection and analysis of yeast centromeric DNA.
    Cell. 1985 Oct;42(3):913-21 PMID: 2996783
  7. GCN4 protein, synthesized in vitro, binds HIS3 regulatory sequences: implications for general control of amino acid biosynthetic genes in yeast.
    Cell. 1985 Nov;43(1):177-88 PMID: 3907851
  8. Nucleotide sequence and transcriptional mapping of the yeast pet56-his3-ded1 gene region.
    Nucleic Acids Res. 1985 Dec 9;13(23):8587-601 PMID: 3001645
  9. Fine-structure analysis of the DNA sequence requirements for autonomous replication of Saccharomyces cerevisiae plasmids.
    Mol Cell Biol. 1986 Jul;6(7):2354-63 PMID: 3023929
  10. Two related regulatory sequences are required for maximal induction of Saccharomyces cerevisiae his3 transcription.
    Mol Cell Biol. 1987 Jan;7(1):104-10 PMID: 3031449
  11. Plasmid vector pBR322 and its special-purpose derivatives--a review.
    Gene. 1986;50(1-3):3-40 PMID: 3034735
  12. Functional inactivation of genes by dominant negative mutations.
    Nature. 1987 Sep 17-23;329(6136):219-22 PMID: 2442619
  13. Visual assay for chromosome ploidy.
    Methods Enzymol. 1987;155:351-72 PMID: 3431466
  14. Direction of chromosome rearrangements in Saccharomyces cerevisiae by use of his3 recombinational substrates.
    Mol Cell Biol. 1988 Oct;8(10):4370-80 PMID: 3054515
  15. Yeast regulatory gene GAL3: carbon regulation; UASGal elements in common with GAL1, GAL2, GAL7, GAL10, GAL80, and MEL1; encoded protein strikingly similar to yeast and Escherichia coli galactokinases.
    Mol Cell Biol. 1988 Aug;8(8):3439-47 PMID: 3062381
  16. Yeast/E. coli shuttle vectors with multiple unique restriction sites.
    Yeast. 1986 Sep;2(3):163-7 PMID: 3333305
  17. Vector systems for the expression, analysis and cloning of DNA sequences in S. cerevisiae.
    Yeast. 1985 Dec;1(2):83-138 PMID: 3916863
  18. Screening lambdagt recombinant clones by hybridization to single plaques in situ.
    Science. 1977 Apr 8;196(4286):180-2 PMID: 322279
  19. Replacement of chromosome segments with altered DNA sequences constructed in vitro.
    Proc Natl Acad Sci U S A. 1979 Oct;76(10):4951-5 PMID: 388424
  20. Sequence of a yeast DNA fragment containing a chromosomal replicator and the TRP1 gene.
    Gene. 1980 Jul;10(2):157-66 PMID: 6248420
  21. Deletion analysis of the Saccharomyces GAL gene cluster. Transcription from three promoters.
    J Mol Biol. 1981 Oct 25;152(2):317-34 PMID: 7035681
  22. Centromeric DNA from Saccharomyces cerevisiae.
    J Mol Biol. 1982 Jun 25;158(2):157-90 PMID: 6750136
  23. Transformation of intact yeast cells treated with alkali cations.
    J Bacteriol. 1983 Jan;153(1):163-8 PMID: 6336730
  24. Nucleotide sequence of yeast LEU2 shows 5'-noncoding region has sequences cognate to leucine.
    Cell. 1982 Dec;31(2 Pt 1):319-25 PMID: 6297759
  25. Expression in Saccharomyces cerevisiae of human interferon-alpha directed by the TRP1 5' region.
    Nucleic Acids Res. 1983 Apr 25;11(8):2287-302 PMID: 6304646
  26. One-step gene disruption in yeast.
    Methods Enzymol. 1983;101:202-11 PMID: 6310324
  27. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity.
    Anal Biochem. 1983 Jul 1;132(1):6-13 PMID: 6312838
  28. DNA sequences of yeast H3 and H4 histone genes from two non-allelic gene sets encode identical H3 and H4 proteins.
    J Mol Biol. 1983 Sep 25;169(3):663-90 PMID: 6355483
  29. Identification of a Ty insertion within the coding sequence of the S. cerevisiae URA3 gene.
    Mol Gen Genet. 1984;193(3):557-60 PMID: 6323928
  30. Structure and function of the yeast URA3 gene: expression in Escherichia coli.
    Gene. 1984 Jul-Aug;29(1-2):113-24 PMID: 6092217
Article Info
Journal
Genetics
Abbr.
Genetics
ISSN
0016-6731
DOI
10.1093/genetics/122.1.19
Published
1989-05-00
Pages
19-27
Language
English
Region
United States
NLM ID
0374636
PMCID
PMC1203683
Subset
IM
External Links
PubMed source DOI: 10.1093/genetics/122.1.19 PMC Full Text View journal details
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