三疣梭子蟹C-JUN氨基末端激酶基因克隆及在病原胁迫后的表达特征分析  

Cloning and Expression Analysis of c-Jun N-Terminal Kinase Gene in Portunus trituberculatus after Pathogenic Stress

作　　者：张云滨 任宪云 高保全[2,3] 吕建建 王磊[1,3] 刘萍 ZHANG Yunbin;REN Xianyun;GAO Baoquan;Lü Jianjian;WANG Lei;LIU Ping(National Demonstration Center for Experimental Fisheries Science Education,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;Yellow Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences,Key Laboratory of Sustainable Development of Marine Fisheries,Ministry of Agriculture and Rural Affairs,Qingdao 266071)

机构地区：[1]上海海洋大学水产科学国家级实验教学示范中心,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]中国水产科学研究院黄海水产研究所农业农村部海洋渔业可持续发展重点实验室,青岛266071

出　　处：《渔业科学进展》2020年第5期92-100,共9页Progress in Fishery Sciences

基　　金：国家虾蟹产业技术体系(CARS-48);国家自然科学基金面上项目(41576147,41876186);泰山领军人才工程高效生态农业创新类计划项目(LJNY2015002);江苏省重点研发计划(面上项目)(BE2017325);中央级公益性科研院所基本科研业务费(20603022018027)共同资助。

摘　　要：C-JUN氨基末端激酶(c-Jun N-terminal kinase,JNK)作为丝裂原活化蛋白激酶(MAPK)超家族的重要一员,在细胞增殖、凋亡和免疫应激等过程中发挥着重要作用。为深入研究JNK基因的免疫防御机制,本研究从三疣梭子蟹(Portunus trituberculatus)转录组数据库中筛选到JNK基因的EST序列,利用RACE扩增技术克隆得到该基因全长序列,命名为PtJNK,cDNA全长为3240 bp,开放阅读框(ORF)为1380 bp,编码459个氨基酸,具有TPY磷酸化位点的S_TKC保守结构域,是JNK基因家族典型特征。组织表达分布结果显示,PtJNK基因在所有组织中均有表达;Real-time PCR检测不同病原刺激下PtJNK基因的表达水平,结果显示,注射副溶血弧菌(Vibrio parahaemolyticus)后,该基因显著下调表达(P<0.05);而注射白斑综合征病毒(WSSV)后,该基因显著上调表达(P<0.05)。综上所述,PtJNK基因是一种广泛表达基因,且在不同的病原感染情况下,该基因的表达模式存在差异,在免疫防御过程中具有重要作用。The c-Jun N-terminal kinase(JNK)is a member of the mitogen-activated protein kinase superfamily,which plays an important role in cell proliferation,apoptosis,and immune stress.To further explore the immune defense mechanism of JNK in Portunus trituberculatus,the EST sequence of PtJNK was isolated from the transcriptome database of P.trituberculatus.In this study,the JNK gene of P.trituberculatus(PtJNK)was successfully cloned by RACE;measurements showed a cDNA length of 3240 bp and an open reading frame of 1380 bp.PtJNK consists of 459 amino acids,including a serine/threonine protein kinase(S-TKc)domain with a conserved Thr-Pro-Tyr(TPY)motif,which is a typical feature of the JNK gene family.The results of tissue expression and distribution show that the PtJNK gene is expressed in all tissues.Real-time PCR was used to detect the expression of PtJNK under different pathogenic stimuli.The results show that expression of the gene is significantly down-regulated after injection of Vibrio parahaemolyticus(P<0.05),while expression is significantly up-regulated when white spot syndrome virus is injected(P<0.05).To sum up,PtJNK gene is a widely expressed gene,and expression of the gene differs according to pathogen.

关 键 词：三疣梭子蟹 PtJNK 基因克隆 病原感染 实时荧光定量PCR 

分 类 号：S917.4[农业科学—水产科学]