Characterization of CLCN5 and Expression Profile under Low-Salinity Stress in Takifugu rubripes
Characterization of CLCN5 and Expression Profile under Low-Salinity Stress in Takifugu rubripes
Lin Ye, Jie-Lan Jiang*, Jia Xu, Rui-Ting Liu, Yin-Qiu Tian, Qing-Qing Zang, Jia-Yi Cao and Ming-Guang Mao*
ABSTRACT
Voltage-gated chloride channel 5 (CLCN5) transports hydrogen ions into the nucleolus and removes chloride ions, which helps the cell to maintain its pH value and osmotic pressure. To reveal the role of CLCN5 under low-salinity stress, a cDNA sequence of CLCN5 (TrCLCN5) was cloned from Takifugu rubripes using the reverse transcription PCR (RT-PCR) technique. The results showed that TrCLCN5 cDNA contained a complete open reading frame of 2505 bp continuously encoding 834 amino acid residues. Sequence alignments revealed that the TrCLCN5 cDNA is highly homologous (96.2% and 96.1%) with that of Takifugu bimaculatus and Takifugu flavidus. SignalP and TMHMM analysis showed that TrCLCN5-deduced protein was a type III transmembrane protein and lacked a typical signal peptide. Conserved Domain analysis revealed that there was a Voltage_CLC domain and two CBS domains located in the TrCLCN5 deduced protein. qPCR analysis showed that TrCLCN5 was highly expressed in the intestine, kidney and liver, and up-regulated in gills under low-salinity stress in 3h to 6h, which indicates that TrCLCN5 may play an important role in the responses to acute low-salinity stress and would be useful in further understanding low-salinity adaption in T. rubripes.
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