Cloning and cold adaptation verification of TRPV1 genes from icefish Chionodraco hamatus and Trematomus bernacchii
WANG Gui-zhi1，ZHANG Yong1，ZHANG Chi1，XU Qiang-hua1，2，3，4，5
1.College of Marine Sciences，ShanghaiOcean University，Shanghai201306，China；2.Key Laboratory of Sustainable Exploitation ofOceanic Fisheries Resources，Ministry of Education，Shanghai201306，China；3.Scientific Observing and Experimental Station ofOceanic Fishery Resources，Ministry of Agriculture，Shanghai 201306，China；4.National Engineering Research Center for Oceanic Fisheries，Shanghai 201306，China；5.Collaborative Innovation Center for Distant-water Fisheries，Shanghai201306，China
Abstract: Transient receptor potential channels V1（TRPV1）gene sequence was cloned in icefish Chionodraco hamatus and Trematomus bernacchii using RT-PCR technology to investigatemolecularmechanism of the two Antarctic fish species to cold adaptation.The bioinformatics analysis indicated that the TRPV1s contained the 2310 bp open reading frame encoding 769 amino acids in length，with relativemolecularweightof88 210 and the theoretical isoelectric point of 7.63 in C.hamatus and relative molecular weight of 88 160 and theoretical isoelectric point of 8.26 in T.bernacchii.Sub-cellular localization revealed that TRPV1 proteins as insoluble transmembrane protein without signal peptide played primrily a role in plasmamembrane and endoplasmic reticulum.Physical and chemical properties showed that there were 34 phosphorylation sites and 6 glycosylation sites in in TRPV1 in C.hamatus and 35 phosphorylation sites，and 5 glycosylation sites in TRPV1 in T.bernacchii，with 6 transmembrane domains and 5 ANK domains.The TRPV1 had secondary structure of random coil.Nucleotide sequence analysis revealed that C.hamatusis TRPV1 had sequence similarity of44.3％-69.8％with TRPV1 in other vertebrates，themaximal similarity（93.4％）with the T.bernacchii TRPV1.Cold adaptation experiments showed that the TRPV1 of both ice fishes was not activated by temperature changes，indicating that the Antarctic fish TRPV1 isnot involved in cold adaptation.
2016, Vol. 31 
