三疣梭子蟹14-3-3基因的克隆及其在低盐和病原胁迫后的表达分析  

Cloning and Expression Analysis of 14-3-3 Gene in Portunus trituberculatus after Exposure to Low Salt and Pathogenic Stress

作　　者：题兴斌 吕建建 宋柳[3] 阎德平 孙东方 刘萍 TI Xingbin;Lü Jianjian;SONG Liu;YAN Deping;SUN Dongfang;LIU Ping(College of Fisheries and Life Science,Shanghai Ocean University,Shanghai 201306;Laboratory for Marine Fisheries Science and Food Production Processes,Pilot National Laboratory for Marine Science and Technology(Qingdao),Qingdao 266071;Key Laboratory of Sustainable Development of Marine Fisheries,Ministry of Agriculture and Rural Affairs/Yellow Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences,Qingdao 266071)

机构地区：[1]上海海洋大学水产与生命学院,上海201306 [2]青岛海洋科学与技术试点国家实验室海洋渔业科学与食物产出过程功能实验室,青岛266071 [3]农业农村部海洋渔业可持续发展重点实验室,中国水产科学研究院黄海水产研究所,青岛266071

出　　处：《渔业科学进展》2021年第1期134-143,共10页Progress in Fishery Sciences

基　　金：农业产业技术体系(CARS-48);泰山领军人才工程高效生态农业创新类计划项目(LJNY2015002);国家自然科学基金面上项目(41776160);山东省重点研发计划(2018GSF121030);中央级公益性科研院所基本科研业务费(20603022018027);江苏省重点研发计划(面上项目)(BE2017325)共同资助。

摘　　要：本研究利用cDNA末端快速扩增技术(RACE)克隆了三疣梭子蟹(Portunus trituberculatus)14-3-3基因(Pt14-3-3)。Pt14-3-3基因cDNA序列全长为2510 bp,包含开放阅读框741 bp,编码由247个氨基酸组成的蛋白,分子量为27.98 kDa。序列比对结果显示,Pt14-3-3基因与中华绒螯蟹14-3-3基因同源性最高(100%)。系统进化树分析表明,Pt14-3-3氨基酸序列与中华绒螯蟹(Eriocheir sinensis)紧密聚为一支。组织表达分析显示,Pt14-3-3基因在肝胰腺、肌肉、鳃、心脏、眼柄、血淋巴中均有表达,其中,在肝胰腺中的表达量最高。低盐胁迫后,Pt14-3-3基因在鳃和肝胰腺中的表达量分别于48 h和12 h显著上调并达到最大值,分别为对照组的1.34倍(P<0.05)和7.54倍(P<0.05)。人工感染副溶血弧菌(Vibrio parahaemolyticus)和白斑综合征病毒(WSSV)后,Pt14-3-3基因在肝胰腺和血细胞中整体均呈显著上调表达,最高上调17.52倍(P<0.05)。本研究结果表明,Pt14-3-3基因可能在三疣梭子蟹低盐适应和免疫反应中发挥重要作用。In this study,the 14-3-3 gene of the swimming crab(Portunus trituberculatus)was cloned using the Rapid Amplification of cDNA Ends(RACE)method.The full length was 2510 bp and the ORF was 741 bp,encoding 247 amino acids and with a predicted molecular weight of 27.98 kDa.Sequence alignment analysis showed that the Pt14-3-3 gene has the highest homology with the 14-3-3 gene of Eriocheir sinensis(100%).Phylogenetic tree analysis indicated that the Pt14-3-3 amino acid sequence of P.trituberculatus was closely clustered into one with Eriocheir sinensis.Tissue expression analysis showed that the Pt14-3-3 gene was expressed in hepatopancreas,muscle,gill,heart,eyestalk,and hemolymph,and the expression level was highest in hepatopancreas.After low salt stress,the expression levels of the Pt14-3-3 gene in sputum and hepatopancreas were significantly upregulated at 48 h and 12 h,respectively,and reached the maximum,which was 1.34 times(P<0.05)and 7.54 times(P<0.05),respectively.After artificial infection with Vibrio parahaemolyticus and WSSV,the Pt14-3-3 gene was upregulated in the hepatopancreas and blood cells and the lowest was up to 17.52 times(P<0.05).The results of this study indicated that the Pt14-3-3 gene plays an important role in low salt adaptation and immune response of the Portunus trituberculatus.

关 键 词：三疣梭子蟹 14-3-3基因 盐度 副溶血弧菌 白斑综合征病毒(WSSV) 

分 类 号：S917[农业科学—水产科学]