Molecular cloning and expression profile of snow trout GPDH gene in response to abiotic stress.
Glycerol-3-phosphate dehydrogenase (GPDH) gene possibly plays a key role for cold acclimation process in snow trout during winter months when water temperature goes down to 4-5 °C. In this study, 1,012 bp nucleotide fragment of GPDH gene was obtained from two snow trout species (Schizothorax richardsonii and S. niger; family: Cyprinidae), distributed in several Himalayan rivers. The gene encoded a protein of 334 amino acids. The encoded protein sequence was very similar to GPDH of Danio rerio (94.36 %) using BLASTx searches. In S. richardsonii the qRT-PCR showed highest expression in muscle tissue followed by liver and also revealed 19 fold gene expression in liver tissue under cold (5 °C) in comparison with warm (15 °C) condition. The elevated expression levels of GPDH cDNA on cold treatment furthermore suggest that GPDH plays a role in stress related responses in S. richardsonii. The phylogenetic analysis showed that the two snow trout species GPDH share the same clade with characterized GPDHs from other teleost fishes suggesting a common evolutionary origin and a similar catalytic function. In addition, the Ka/Ks ratios of these sequences suggested that they are under purifying selection. Moreover, the expression profile of GPDH gene among co generic species of genus Schizothorax showed that GPDH cDNA expression was highest in S. richardsonii and lowest in S. esocinus which gives an indication of species specific adaptation in relation to different geographical areas.
Molecular Genetics Laboratory, Directorate of Coldwater Fisheries Research, Indian Council of Agricultural Research, Bhimtal, Nainital, 263136, Uttarakhand, India. abarat58@hotmail.com
Ashoktaru
A
Barat
Chirag
C
Goel
Ankita
A
Tyagi
Shahnawaz
S
Ali
Prabhati K
PK
Sahoo
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution / Q000235:genetics; D000818:Animals; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics / Q000302:isolation & purification; D020869:Gene Expression Profiling; D015971:Gene Expression Regulation, Enzymologic; D005990:Glycerol / Q000097:blood; D005993:Glycerolphosphate Dehydrogenase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D010802:Phylogeny; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D012914:Snow; D013312:Stress, Physiological / Q000235:genetics*; D013696:Temperature; D014337:Trout / Q000097:blood / Q000235:genetics* / Q000502:physiology*
D018076:DNA, Complementary; D005993:Glycerolphosphate Dehydrogenase; D005990:Glycerol