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

Mol Cell Biol. 1983 Apr;3(4):580-6. doi: 10.1128/mcb.3.4.580-586.1983.

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*

Substances

  • DNA, Recombinant
  • Alcohol Oxidoreductases
  • 3-Isopropylmalate Dehydrogenase
  • Galactosidases
  • beta-Galactosidase
  • Leucine