MOLECULAR CLONING, CHARACTERIZATION, AND EXPRESSION OF AQUAPORIN1 GENE IN CRASSOSTREA HONGKONGENSIS
Author(s): WAN Qian, ZHANG Yang, ZHANG Yue-Huan, YU Zi-Niu, Key Laboratory of Tropical Marine Bio-resources and Ecology and Guangdong Provincial Key Laboratory of Applied Marine Biology, South China Sea Institute of Oceanology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, South China Sea Bio-resource Exploitation and Utilization Collaborative Innovation Center, University of Chinese Academy of Sciences
Pages: 1078-1087
Year: 2015 Issue: 5
Journal: Oceanologia et Limnologia Sinica
Keyword: Crassostrea hongkongensis; Aquaporin1; salinity; Real-Time PCR;
Abstract: Aquaporin1(AQP1) is one of water channel proteins that facilitate water and/or small solutes across cell membrane responding to osmotic gradients. We cloned the AQP1 gene by RT-PCR and RACE technology in mollusk Crassostrea hongkongensis and named Ch AQP1(Gen Bank accession number: KJ704847). The full-length c DNA of Ch AQP1 is 1153 bp in length with ORF of 888 bp, encoding a peptide of 295 amino acids. Ch AQP1 includes a conserved MIP domain, six transmembrane regions, five loops, two NPA boxes, and an ar/R(aromatic/arginine) selectivity filter. Phylogenetic analysis showed that Ch AQP1 belongs to the AQP1-like subfamily. Tissue distribution of Ch AQP1 m RNA indicates that Ch AQP1 is constitutively expressed in all detected tissues, especially in adductor muscle and mantle. Furthermore, as shown in the expression pattern in gill under osmotic stress revealed in real-time PCR analysis, Ch AQP1 m RNA stayed almost unchanged in hypo-osmotic exposure, whereas its m RNA level was significantly down-regulated on Days1(P<0.01), 3, and 5(P<0.05) upon hyper-osmotic exposure. Thus, the expression pattern of Ch AQP1 m RNA in gill is inducible under salinity stress, which strongly discloses the involvement of the gene in the regulation of osmotic homeostasis in C. hongkongensis. With these basic molecular findings, future works shall focus on the mechanism of euryhaline adaptation in C. hongkongensis.
Pages: 1078-1087
Year: 2015 Issue: 5
Journal: Oceanologia et Limnologia Sinica
Keyword: Crassostrea hongkongensis; Aquaporin1; salinity; Real-Time PCR;
Abstract: Aquaporin1(AQP1) is one of water channel proteins that facilitate water and/or small solutes across cell membrane responding to osmotic gradients. We cloned the AQP1 gene by RT-PCR and RACE technology in mollusk Crassostrea hongkongensis and named Ch AQP1(Gen Bank accession number: KJ704847). The full-length c DNA of Ch AQP1 is 1153 bp in length with ORF of 888 bp, encoding a peptide of 295 amino acids. Ch AQP1 includes a conserved MIP domain, six transmembrane regions, five loops, two NPA boxes, and an ar/R(aromatic/arginine) selectivity filter. Phylogenetic analysis showed that Ch AQP1 belongs to the AQP1-like subfamily. Tissue distribution of Ch AQP1 m RNA indicates that Ch AQP1 is constitutively expressed in all detected tissues, especially in adductor muscle and mantle. Furthermore, as shown in the expression pattern in gill under osmotic stress revealed in real-time PCR analysis, Ch AQP1 m RNA stayed almost unchanged in hypo-osmotic exposure, whereas its m RNA level was significantly down-regulated on Days1(P<0.01), 3, and 5(P<0.05) upon hyper-osmotic exposure. Thus, the expression pattern of Ch AQP1 m RNA in gill is inducible under salinity stress, which strongly discloses the involvement of the gene in the regulation of osmotic homeostasis in C. hongkongensis. With these basic molecular findings, future works shall focus on the mechanism of euryhaline adaptation in C. hongkongensis.
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