文章摘要
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
投稿时间:2019-06-30  修订日期:2019-07-29
DOI:
中文关键词: 三疣梭子蟹  PtJNK  基因克隆  病原感染  实时荧光定量
英文关键词: Portunus trituberculatus  PtJNK  Gene cloning  Pathogenic infection  Quantitative Real-time PCR
基金项目:国家自然科学基金面上项目(41876186,41576147)、泰山领军人才工程高效生态农业创新类计划项目(LJNY2015002)、国家虾蟹产业技术体系(CARS-48)、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
作者单位E-mail
张云滨 上海海洋大学 930476134@qq.com 
任宪云 黄海水产研究所  
高保全 黄海水产研究所  
吕建建 黄海水产研究所  
王磊 上海海洋大学  
刘萍 黄海水产研究所 liuping@ysfri.ac.cn 
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中文摘要:
      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 3240bp,开放阅读框(ORF)1380bp,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,其中胃和肌肉中的表达量最高;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),而注射白斑综合症病毒(WSSV)后该基因显著上调表达(P<0.05)。综上所述:在不同的病原感染情况下,PtJNK基因的表达模式存在着差异,在免疫防御过程中具有重要作用。
英文摘要:
      The c-Jun N-terminal kinase (JNK) is a member of mitogen-activated protein kinases (MAPK) superfamily, which plays an important role in cell proliferation, apoptosis and immune stress. In order to further explore the immune defense mechanism of JNK in P.trituberculatus, the EST sequence of PtJNK is isolated from the transcriptome database of P. trituberculatus. the PtJNK gene is successfully cloned by RACE in this study, cDNA 3240 bp, and the open reading frame (ORF) is 1380 bp, The PtJNK consists of 459 amino acids, which includes a serine/threonine protein kinase (S-TKc) domain with a conserved Thr-Pro-Tyr(TPY) motif, it is a typical feature of JNK gene family. The results of tissue expression and distribution show that PtJNK gene is expressed in all tissues, especially in stomach and muscle. Real-time PCR is used to detect the expression of PtJNK under different pathogenic stimuli. The results show that the expression of PtJNK gene is significantly down-regulated after injection of Vibrio parahaemolyticus (P<0.05), while the expression of PtJNK gene is significantly up-regulated when WSSV is injected (P<0.05). Therefore, we speculates that Vibrio parahaemolyticus injection inhibites the expression of PtJNK, and the expression patterns of PtJNK are different under different pathogenic infections, which plays an important role in the immune process.
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