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The 3-phosphoglycerate kinase gene of the yeast Yarrowia lipolytica de-represses on gluconeogenic substrates
- Le Dall, M.T., Nicaud, J.M., Treton, B.Y., Gaillardin, C.M.
- Current genetics 1996 v.29 no.5 pp. 446-456
- Saccharomycetales, recombinant DNA, site-directed mutagenesis, promoter regions, lactic acid, structural genes, open reading frames, glucose, gene expression, citric acid, introns, messenger RNA, phosphoglycerate kinase, pyruvic acid, complementary DNA, glycerol, beta-galactosidase, amino acid sequences, enzyme activity, culture media, nutrient availability, reporter genes, mutants
- We have isolated the 3-phosphoglycerate kinase (PGK) gene of the yeast Yarrowia lipolytica by probing a genomic library with a PCR fragment amplified with primers deduced from two highly conserved regions of various PGKs. It is a unique sequence encoding a polypeptide of 417 residues with extensive homology to other PGKs, especially to that of Aspergillus nidulans (76% identity). The expression of the Y. lipolytica PGK1 gene proved to be higher on gluconeogenic substrates than on glycolytic ones. Haploid strains harboring a disrupted allele were able to grow on mixtures of a gluconeogenic carbon source and of a glycolytic one, but required proline supplementation in the presence of glucose, and were inhibited by glycerol.