Cloning and characterization of a glycerol-3-phosphate dehydrogenase (NAD+) gene from the halotolerant yeast Pichia farinosa.
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.
Peng
Feng
F
Key Laboratory of Bio-Resources and Eco-Environment of Ministry of Education, College of Life Science, Sichuan University, Chengdu 610064, People's Republic of China.
Li
Gang
G
Wang
Xiaoxing
X
Jiang
Yan
Y
Yang
Yi
Y
D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary; D004271:DNA, Fungal / Q000235:genetics; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D018628:Gene Dosage; D015966:Gene Expression Regulation, Fungal; D005800:Genes, Fungal; D005816:Genetic Complementation Test; D050536:Glycerol-3-Phosphate Dehydrogenase (NAD+) / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D009994:Osmolar Concentration; D010843:Pichia / Q000201:enzymology* / Q000235:genetics*; D011401:Promoter Regions, Genetic; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D012965:Sodium Chloride / Q000494:pharmacology; D013312:Stress, Physiological
D018076:DNA, Complementary; D004271:DNA, Fungal; D005656:Fungal Proteins; D012965:Sodium Chloride; D050536:Glycerol-3-Phosphate Dehydrogenase (NAD+)