【作者】 李婵； 姚翠鸾； 王志勇；

【Author】 Chan Li,Cui-Luan Yao~*,Zhi-Yong Wang (Key Laboratory of Helthy Mariculture for East China Sea,Ministry of Agriculture of the P.R.C,Fisheries College, Jimei University,Xiamen,361021,China)

【机构】 集美大学农业部东海海水健康养殖重点实验室； 集美大学水产学院；

【摘要】 本研究利用RT-PCR和RACE-PCR技术克隆了大黄鱼IL-8基因全长cDNA序列,分析了其分子结构,并利用Real-Time PCR技术研究了该基因在大黄鱼不同组织的表达谱,以及溶藻弧菌(Vibrio alginolyticus)和病原模式分子LPS、poly I:C刺激后头肾、脾脏和肝脏组织中该基因在转录水平的表达变化。结果表明:大黄鱼IL-8基因全长为931bp,包括118bp的5’端非翻译区,528bp的3’端非翻译区,285bp的开放阅读框,编码94个氨基酸。3’端非翻译区包含1个RNA不稳定基序(ATTTA),加尾信号(AATAAA)在Poly A尾上游19bp处。预测IL-8氨基酸序列分子量为10.524kDa,等电点为9.64;其N末端具有20个氨基酸的信号肽序列,信号肽后的氨基酸残基包含一个SCY结构域。IL-8在大黄鱼的大部分组织中都有表达,其中在肝脏和皮肤组织中表达量较高,在脾脏中的表达量较低;LPS、polyI:C刺激可以诱导IL-8基因在头肾和脾脏中表达量增加,溶藻弧菌刺激诱导头肾中IL-8基因的表达量上调,但三这种刺激均导致IL-8在肝脏中的转录表达下调更多还原

【Abstract】 The full length cDNA of Interleukin-8(IL-8) was cloned from Larimichthys crocea by reverse transcription polymerase chain reaction(RT-PCR) and rapid amplification of cDNA ends (RACE) PCR.In addition,the tissue expression and temporal expression profiles of IL-8 gene after the stimulation with LPS,poly I:C and pathogenic bacteria Vibrio alginolyticus were discribed.The full-length cDNA of IL-8 was 931 bp,containing a 118 bp 5’-untranslated region (UTR),a 528bp 3’- UTR and a 285 bp open reading frame(ORF) which encoded 94 amino acids. The putative molecular weight of IL-8 is 10.524 kDa and theoretical isoelectric point is 9.64.A signal peptide including 20 amino acid residues at N-terminal and a SCY domain was predicted in IL-8.Real-time PCR analysis indicated that a broad expression of IL-8 in the most of tissues,with the predominant expression in the liver and skin,and the weakest expression in spleen.After injection with LPS and poly I:C,IL-8 expression level showed increase in head-kidney and spleen, and V.alginolyticus could induce IL-8 transcript up-regulation in head-kidney.But IL-8 transcript down-regulated in the liver after the three stimulations.更多还原