Cloning of a eukaryotic regulatory gene

Mol Gen Genet. 1981;184(3):394-9. doi: 10.1007/BF00352511.

Abstract

From a pool of hybrid plasmids carrying Sau3A fragments representing the entire yeast (S. cerevisiae) genome, a DNA fragment containing the regulatory gene PPRI was cloned by complementation of a non-inducible ppr1 mutation which confers to the cells an increased sensitivity to 6-azauracil. Cells containing the cloned DNA regained the ability to induce the synthesis of URA1 and URA3 gene products controlled by PPR1. A physical map has been constructed and the study of subcloned restriction endonuclease fragments from the original yeast DNA fragment allowed us to localize the wile-type PPR1 regulatory gene within a 3 kilobase-pair region. The ppr1 RNA level was measured and the hybridization data indicate in a wild-type strain a low efficiency of transcription of PPR1 as compared to the structural URA3 gene, without effect of inducing conditions.

MeSH terms

  • Base Sequence
  • Cloning, Molecular*
  • DNA
  • DNA Restriction Enzymes
  • Dihydroorotate Oxidase / genetics
  • Escherichia coli / genetics*
  • Genes, Regulator*
  • Mutation
  • Orotidine-5'-Phosphate Decarboxylase / genetics
  • Plasmids
  • Saccharomyces cerevisiae / genetics*
  • Transcription, Genetic

Substances

  • DNA
  • Dihydroorotate Oxidase
  • DNA Restriction Enzymes
  • Orotidine-5'-Phosphate Decarboxylase