title: Cloning and characterization of the orotidine-5'-phosphate decarboxylase gene (URA3) from the osmotolerant yeast Candida magnoliae.
abstract: We determined the nucleotide sequence of the URA3 gene encoding orotidine-5'-phosphate decarboxylase (OMPDCase) of the erythritol-producing osmotolerant yeast Candida magnoliae by degenerate polymerase chain reaction and genome walking. Sequence analysis revealed the presence of an uninterrupted open-reading frame of 795 bp, encoding a 264 amino acid residue protein with the highest identity to the OMPDCase of the yeast Kluyveromyces marxianus. Phylogenetic analysis of the deduced amino acid sequence revealed that it shared a high degree of identity with other yeast OMPDCase homologs. The cloned URA3 gene successfully complemented the ura3 null mutation in Saccharomyces cerevisiae, revealing that it encodes a functional OMPDCase in C. magnoliae. An enzyme activity assay and reverse transcription polymerase chain reaction indicated that the expression level of the C. magnoliae URA3 gene in S. cerevisiae was not as high as that of the S. cerevisiae URA3 gene. The GenBank accession number for C. magnoliae URA3 is JF521441.
authors.affiliation: School of Biotechnology and Bioengineering, Kangwon National University, Chuncheon 200-701, Korea.
authors.forename: Eun-Hee
authors.initials: EH
authors.lastname: Park
authors.forename: Jin-Ho
authors.initials: JH
authors.lastname: Seo
authors.forename: Myoung-Dong
authors.initials: MD
authors.lastname: Kim
mesh_terms: D000595:Amino Acid Sequence; D002175:Candida / Q000737:chemistry / Q000145:classification / Q000201:enzymology* / Q000235:genetics; D003001:Cloning, Molecular; D005656:Fungal Proteins / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D009964:Orotidine-5'-Phosphate Decarboxylase / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D009995:Osmosis; D010802:Phylogeny; D016415:Sequence Alignment
chemical_list: D005656:Fungal Proteins; D009964:Orotidine-5'-Phosphate Decarboxylase