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PMID: 6406836 Published · ppublish English Journal Article

Fusion of the Saccharomyces cerevisiae leu2 gene to an Escherichia coli beta-galactosidase gene.

Molecular and cellular biology ·Vol. 3 ·No. 4 ·1983-04-00 ·Pages 580-6

Martinez-Arias AE, Casadaban MJ

Abstract

The promoter and translation initiation region of the Saccharomyces cerevisiae leu2 gene was fused to the Escherichia coli beta-galactosidase gene. This fusion located the control region of the leu gene and orientated its direction of expression. When the fusion was placed into yeast cells, beta-galactosidase was expressed under the same regulatory pattern as the original leu2 gene product: its synthesis was repressed in the presence of leucine and threonine. Sensitive chromogenic substrates for beta-galactosidase were used to detect expression in isolated colonies growing on agar medium. Mutant yeast cells with increased beta-galactosidase activity were identified by the color of the colonies they formed. One class of mutants obtained appeared to affect ars1 plasmid maintenance, and another class appeared to affect beta-galactoside uptake.

MeSH Terms
3-Isopropylmalate Dehydrogenase Alcohol Oxidoreductases/genetics DNA, Recombinant Escherichia coli/genetics Galactosidases/genetics Gene Expression Regulation Leucine/genetics Mutation Plasmids Saccharomyces cerevisiae/genetics beta-Galactosidase/genetics
Chemicals
DNA, Recombinant Alcohol Oxidoreductases 3-Isopropylmalate Dehydrogenase Galactosidases beta-Galactosidase Leucine
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Martinez-Arias A E
Casadaban M J
References (22)
22 references, click to expand
  1. Genetical mutants induced by ethyl methanesulfonate in Saccharomyces.
    Can J Genet Cytol. 1965 Sep;7(3):491-9 PMID: 5866861
  2. Biosynthesis of branched-chain amino acids in yeast: regulation of leucine biosynthesis in prototrophic and leucine auxotrophic strains.
    J Bacteriol. 1968 Dec;96(6):2018-24 PMID: 5724970
  3. Lac repressor can be fused to beta-galactosidase.
    Nature. 1974 Jun 7;249(457):561-3 PMID: 4599764
  4. Transposition and fusion of the lac genes to selected promoters in Escherichia coli using bacteriophage lambda and Mu.
    J Mol Biol. 1976 Jul 5;104(3):541-55 PMID: 781293
  5. Functional expression of cloned yeast DNA in Escherichia coli.
    Proc Natl Acad Sci U S A. 1977 Feb;74(2):487-91 PMID: 322128
  6. Transformation of yeast.
    Proc Natl Acad Sci U S A. 1978 Apr;75(4):1929-33 PMID: 347451
  7. Transformation of yeast by a replicating hybrid plasmid.
    Nature. 1978 Sep 14;275(5676):104-9 PMID: 357984
  8. Genetic fusions for operon analysis.
    Annu Rev Genet. 1978;12:193-221 PMID: 371524
  9. Lactose genes fused to exogenous promoters in one step using a Mu-lac bacteriophage: in vivo probe for transcriptional control sequences.
    Proc Natl Acad Sci U S A. 1979 Sep;76(9):4530-3 PMID: 159458
  10. Sterile host yeasts (SHY): a eukaryotic system of biological containment for recombinant DNA experiments.
    Gene. 1979 Dec;8(1):17-24 PMID: 395030
  11. Analysis of gene control signals by DNA fusion and cloning in Escherichia coli.
    J Mol Biol. 1980 Apr;138(2):179-207 PMID: 6997493
  12. Expression of a foreign eukaryotic gene in Saccharomyces cerevisiae: beta-galactosidase from Kluyveromyces lactis.
    Gene. 1980 Sep;10(4):347-56 PMID: 6253356
  13. Transposed LEU2 gene of Saccharomyces cerevisiae is regulated normally.
    J Bacteriol. 1980 Nov;144(2):852-5 PMID: 7000755
  14. In vitro gene fusions that join an enzymatically active beta-galactosidase segment to amino-terminal fragments of exogenous proteins: Escherichia coli plasmid vectors for the detection and cloning of translational initiation signals.
    J Bacteriol. 1980 Aug;143(2):971-80 PMID: 6162838
  15. Yeast genes fused to beta-galactosidase in Escherichia coli can be expressed normally in yeast.
    Proc Natl Acad Sci U S A. 1981 Apr;78(4):2460-4 PMID: 6787605
  16. 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 PMID: 6796588
  17. The primary structure of the Saccharomyces cerevisiae gene for alcohol dehydrogenase.
    J Biol Chem. 1982 Mar 25;257(6):3018-25 PMID: 6277922
  18. DNA sequence required for efficient transcription termination in yeast.
    Cell. 1982 Mar;28(3):563-73 PMID: 6280875
  19. Efficient expression of Escherichia coli galactokinase gene in mammalian cells.
    Proc Natl Acad Sci U S A. 1982 Jan;79(2):257-61 PMID: 6281760
  20. Regulation of HIS4-lacZ fusions in Saccharomyces cerevisiae.
    Mol Cell Biol. 1982 Oct;2(10):1212-9 PMID: 6817079
  21. 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
  22. Beta-galactosidase gene fusions for analyzing gene expression in escherichia coli and yeast.
    Methods Enzymol. 1983;100:293-308 PMID: 6312261
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
Published
1983-04-00
Pages
580-6
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC368573
Subset
IM
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