Cloning and characterization of a glycerol-3-phosphate dehydrogenase (NAD+) gene from the halotolerant yeast Pichia farinosa

Abstract

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.