Cloning of a eukaryotic regulatory gene.
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.
D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA; D004262:DNA Restriction Enzymes; D004081:Dihydroorotate Oxidase / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics*; D005809:Genes, Regulator; D009154:Mutation; D009964:Orotidine-5'-Phosphate Decarboxylase / Q000235:genetics; D010957:Plasmids; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D014158:Transcription, Genetic
D004247:DNA; D004081:Dihydroorotate Oxidase; D004262:DNA Restriction Enzymes; D009964:Orotidine-5'-Phosphate Decarboxylase