中国明对虾Imd免疫信号通路相关基因克隆及表达分析  

Cloning of Imd Immune Signal Pathway Relating Genes of Fenneropenaeus chinensis and Their Expression Analysis

作　　者：葛倩倩[1,2] 李健[2] 梁俊平[1,2] 刘萍[2] 赵法箴[2] 潘鲁青[1] 刘德月[2] 

机构地区：[1]中国海洋大学,山东青岛266003 [2]中国水产科学研究院黄海水产研究所,山东青岛266071

出　　处：《中国海洋大学学报（自然科学版）》2014年第1期46-51,共6页Periodical of Ocean University of China

基　　金：国家虾产业技术体系项目(CARS-47);国家高技术研究发展计划项目(2012AA10A409);公益性行业(农业)科研专项费用项目(201103034)资助

摘　　要：Imd免疫信号通路在无脊椎动物抗革兰氏阴性菌方面起到十分重要的作用。在已知中国明对虾Imd通路下游核转录因子Relish基因全长的情况下,利用同源克隆方法获得了Imd通路上游基因Imd的cDNA序列,命名为FcImd,该基因全长783bp,5’非编码区45bp,3’非编码区255bp,开放阅读框483bp,编码160个氨基酸,包括一个位于C端的死亡结构域。同源性分析表明,FcImd基因氨基酸序列与其他甲壳动物的氨基酸序列高度保守,与凡纳滨对虾和日本囊对虾的同源性分别为99%和88%。于鳗弧菌感染后的1,3,6,12,24,48,96,144h记录实验组和对照组的累积死亡率,并分别取血淋巴用于检测中国明对虾Imd及Relish基因在不同时间点的表达量变化情况。结果显示:鳗弧菌感染后,实验组Imd和Relish表达量均显著高于对照组(P<0.05),其中Imd表达量呈先升高后降低趋势,Relish表达量在1h时达到最高,然后呈逐渐降低趋势。中国明对虾感染鳗弧菌后Imd和Relish基因呈不同步上调表达,说明Imd和Relish基因的表达与弧菌刺激密切相关,提示Imd和Relish基因参与了对虾抗菌反应的Imd信号转导通路。Fenneropenaeus chinensis is an important mariculture species in China. Recently, epidemic prawn disease as the main factor has limited its sustainable and healthy culture. Invertebrates use their in- nate immunity as the the first barrier against pathoghens. The inducible antimicrobial peptides （AMPs） regulated by Toll and immune deficiency （Imd） pathway are effectors of Drosophila innate immunity, and Imd pathway responds to the infection of Gram-negative bacteria. In this study, we cloned an Imd gene from F. chinensis, named Fclmd. The full-length cDNA of FcImd was 783 bp, consisting of an ORF of 483 bp, which encoded putatively a protein of 160 amino acids with a death domain at its C-terminal. Ho- mology analysis revealed that FcImd was highly conservative among crustaceans. The similarity of FcImd with that of Litopenaeus vannamei and Marsupenaeus japonicus were 99% and 88%, respectively. Phylo- genetic analysis showed that FcImd clustered with insect IMDs and vertebrates RIP1s. The hemocytes were drawn at 1,3,6,12,24,48,96 and 144 h after being challenged with Vibrio anguillarum. The expres- sion level of FcImd in hemocytes was analyzed with quantitative real-time PCR. It was found that the ex- pression of FcImd and Relish was up-regulated to different extents. Our findings implied that Imd immune signal pathway may play an important role in prawn immune response.

关 键 词：中国明对虾 Imd免疫信号通路 Imd基因 Relish基因 克隆 表达 

分 类 号：Q786[生物学—分子生物学] S917[农业科学—水产科学]