Cloning of a eukaryotic regulatory gene

✍ Scribed by Losson, Régine ;Lacroute, François

• Book ID: 104735864
• Publisher: Springer
• Year: 1981
• Tongue: English
• Weight: 754 KB
• Volume: 184
• Category: Article
• ISSN: 0026-8925
• DOI: 10.1007/bf00352511

✦ Synopsis

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 pprl 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 wild-type PPR1 regulatory gene within a 3 kilobase-pair region. The pprl 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.