三疣梭子蟹aqp2基因克隆及其在蜕皮中的功能

CLONING OF AQP2 GENE OF PORTUNUS TRITUBERCULATUS AND ITS FUNCTION IN MOLTING

为研究水通道蛋白AQP的生理功能,克隆了三疣梭子蟹(Portunus trituberculatus)水通道蛋白2基因(ptaqp2),该基因全长4126 bp,编码522个氨基酸,具有水通道蛋白基因家族的保守结构域及功能结构域;进化树聚类分析结果显示, ptaqp2基因属于水通道蛋白基因家族C-AQP类基因;组织表达结果显示, ptaqp2呈泛组织表达特征,在眼柄中的表达量最高,在肠和肌肉中也有较高表达,在血淋巴中的表达最低;ptaqp2基因在不同蜕皮时期呈显著差异表达(P<0.05),且在蜕皮前期表达量最高;在蜕皮激素(Molting Hormone简称MH)刺激下,该基因在肌肉、肠及胃中均呈不同程度的上调表达趋势;通过RNAi技术在蜕皮前期敲降ptaqp2的表达,发现能够显著推迟蜕皮过程。研究初步证明了ptaqp2基因在三疣梭子蟹蜕皮中发挥重要作用。

In order to study the physiological function of aquaporin AQP,the aquaporin 2 gene(ptaqp2)of Portunus trituberculatus was cloned,which is full-length 4126 bp,encodes 522 amino acids and has the conserved domain and functional domain of aquaporin gene family.Cluster analysis showed that ptaqp2 gene belonged to C-AQP gene of aquaporin gene family.The results of tissue expression showed that the expression of ptaqp2 was the highest in eye stalk,the highest in intestine and muscle,and the lowest in hemolymph.The expression of ptaqp2 gene was significantly different in different molting stages,and the highest expression was in the early stage of molting.Under the stimulation of Molting Hormone(MH),the expression of ptaqp2 gene was up-regulated in muscle,intestine and stomach.By knocking down the expression of ptaqp2 in the early stage of molting by RNAi technique,it was found that it could significantly delay the molting process.It has been preliminarily proved that ptaqp2 gene plays an important role in molting of Portunus trituberculatus.