Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Cloning and Characterization of the Orotidine-5'-Phosphate Decarboxylase Gene (URA3) from the Osmotolerant Yeast Candida magnoliae

  • Park, Eun-Hee (School of Biotechnology and Bioengineering, Kangwon National University) ;
  • Seo, Jin-Ho (Department of Agricultural Biotechnology, Seoul National University) ;
  • Kim, Myoung-Dong (School of Biotechnology and Bioengineering, Kangwon National University)
  • Received : 2011.11.28
  • Accepted : 2012.01.11
  • Published : 2012.05.28
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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.

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References

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