| 摘要: |
| 水通道蛋白1(Aquaporin1, AQP1)是一类通过渗透梯度将水或一些小的中性分子快速穿过细胞膜的通道蛋白。通过RT-PCR和RACE技术克隆获得了香港牡蛎AQP1基因全长,并命名为ChAQP1(GenBank登录号:KJ704847)。该基因全长1153bp,开放阅读框长度为888bp,编码295个氨基酸残基。ChAQP1基因包括1个保守的MIP结构域、6个跨膜区、5个连接环、2个NPA(Asn-Pro-Ala)基序和选择性水孔构件ar/R(aromatic/arginine)。系统学分析表明ChAQP1属于AQP1-like型水通道蛋白。mRNA组织分布结果显示, ChAQP1在各个组织中均有表达,其中在闭壳肌和外套膜中表达量相对较高。利用实时定量PCR分析高、低盐胁迫下其在鳃中的表达模式,结果表明, ChAQP1 mRNA表达量在低盐处理下基本没有太大变化;在高盐胁迫下,第1天(P<0.01)、第3天和第5天(P<0.05)显著下调;这说明ChAQP1基因参与了香港牡蛎的渗透压平衡调节。 |
| 关键词: 香港牡蛎 水通道蛋白1 盐度 实时定量PCR |
| DOI：10.11693/hyhz20140400127 |
| 分类号: |
| 基金项目:广东省海洋与渔业局重点项目资助,A201501B03号;国家自然科学基金项目资助,2010CB126404号;NSFC-广东联合基金项目资助,U1201215号。 |
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| MOLECULAR CLONING, CHARACTERIZATION, AND EXPRESSION OF AQUAPORIN1 GENE IN CRASSOSTREA HONGKONGENSIS |
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WAN Qian1,2,3, ZHANG Yang1,2, ZHANG Yue-Huan1,2, YU Zi-Niu1,2
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1.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, Guangzhou 510301, China;2.South China Sea Bio-resource Exploitation and Utilization Collaborative Innovation Center, Guangzhou 510275, China;3.University of Chinese Academy of Sciences, Beijing 100049, China
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| 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 ChAQP1(GenBank accession number:KJ704847). The full-length cDNA of ChAQP1 is 1153bp in length with ORF of 888bp, encoding a peptide of 295 amino acids. ChAQP1 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 ChAQP1 belongs to the AQP1-like subfamily. Tissue distribution of ChAQP1 mRNA indicates that ChAQP1 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, ChAQP1 mRNA stayed almost unchanged in hypo-osmotic exposure, whereas its mRNA 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 ChAQP1 mRNA 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. |
| Key words: Crassostrea hongkongensis Aquaporin1 salinity Real-Time PCR |
