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Cloning and characterization of a glycerol‐3‐phosphate dehydrogenase ( NAD + ) gene from the halotolerant yeast Pichia farinosa
Author(s) -
Peng Feng,
Li Gang,
Wang Xiaoxing,
Jiang Yan,
Yang Yi
Publication year - 2010
Publication title -
yeast
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 102
eISSN - 1097-0061
pISSN - 0749-503X
DOI - 10.1002/yea.1736
Subject(s) - biology , open reading frame , microbiology and biotechnology , dehydrogenase , complementary dna , gene , biochemistry , yeast , glycerol 3 phosphate dehydrogenase , molecular cloning , enzyme , peptide sequence
In this study, a novel glycerol‐3‐phosphate dehydrogenase (NAD + ) (EC1.1.1.8) gene ( PfGPD ) was cloned from halotolerant yeast Pichia farinosa , using degenerate reverse transcription (RT)–PCR and rapid amplification of cDNA ends (RACE) methods. The full‐length cDNA of the PfGPD gene is 1403 bp, which has an open reading frame (ORF) encoding 370 amino acids. PfGPD is conserved with other glycerol‐3‐phosphate dehydrogenase genes and presents a single copy in the P. farinosa genome. Transcriptional analysis revealed that PfGPD gene expression level was high after 2 h induction in a hyperosmotic environment containing 2 M NaCl and returned to normal within 6 h. These results suggest that the PfGPD gene is induced by salt stress. Yeast complementation experiment indicated that PfGPD complements gpd1 mutation in S. cerevisiae . The Accession No. for PfGPD in GenBank is EF601986. Copyright © 2009 John Wiley & Sons, Ltd.