长毛明对虾酚氧化酶原基因cDNA克隆及组织表达分析  

cDNA Cloning and Tissue Expression of Prophenoloxidase Gene in Fenneropenaeus penicillatus

作　　者：韦光本 伍元植 张依琳 吴凡 黄郁葱[1] 蔡双虎[1] 汤菊芬[1] Wei Guangben;Wu Yuanzhi;Zhang Yilin;Wu Fan;Huang Yucong;Cai Shuanghu;Tang Jufen(Guangdong Key Laboratory of Pathogenic Biology and Epidemiology of Aquatic Economic Animals,Fisheries College of Guangdong Ocean University,Zhanjiang,524088)

机构地区：[1]广东海洋大学水产学院广东省水产经济动物病原生物学及流行病学重点实验室

出　　处：《基因组学与应用生物学》2019年第11期4910-4918,共9页Genomics and Applied Biology

基　　金：广东省海洋和渔业发展专项资金(鱼病防治)项目;广东海洋大学水产学院本科生进实验室项目共同资助

摘　　要：酚氧化酶(phenoloxidase,PO)属于对虾免疫系统的核心酶,在机体抵抗微生物感染过程中起到重要的作用。本研究利用RT-PCR和RACE技术克隆长毛明对虾(Fenneropenaeus penicillatus)酚氧化酶原(prophenoloxidase,proPO)基因(F-proPO)。cDNA全长3003 bp,包含63 bp的5′-UTR、879 bp的3′-UTR和编码686个氨基酸的2061 bp的开放阅读框(ORF),预测分子量为78.05 kD,理论等电点(pI)为5.79;含有3个串联式血蓝蛋白结构功能域、2个保守的Cu2+结合位点和6个组氨酸残基;多重比对结果表明,F-proPO与中国明对虾proPO相似度最高(98%),与其他甲壳类动物proPO相似度为51%~93%。实时荧光定量PCR检测结果表明,该基因的表达具有组织特异性,在血淋巴中表达量最高。人工感染溶藻弧菌后,该基因在鳃、血淋巴、肠和性腺中表达水平显著上调;人工感染白斑综合征病毒后,该基因在脑、鳃、心、血淋巴和肠中表达水平显著上调。可见,长毛明对虾PO是参与机体免疫防御反应的一种重要分子。Phenoloxidase(PO)belongs to the core enzyme of Penaeus chine ns is immune system and plays an important role in the immune system of shrimp in the host’s resistance to microbial infection.The prophenoloxidase(proPO)gene of Fenneropenaeus penicillatus(F-proPO)was cloned by RT-PCR and RACE technology in this study.The full-length cDNA was 3003 bp,including a 63 bp of 5’-UTR,a 879 bp of 3’-UTR and a 2061 bp of open reading frame(ORF)encoded a protein of 686 amino acids,the molecular weight and theoretical pI of the deduced F-proPO protein is 78.05 kD and 5.79,respectively.The F-proPO protein contains three tandem limpet hemocyanin domains,two conserved copper ion binding sites and six histidine residues.Multiple alignments showed that F-proPO had the highest similarity with that of F.chinensis(98%),similarity with other crustaceans proPO is 51%~93%.The real-time quantitative PCR results showed that the expression of F-proPO gene is tissue-specific,with the highest expression level in the hemolymph.And the expression levels of F-proPO were significantly upregulated in gill,hemolymph,intestine and gonad after infected by Vibrio alginolyticus,and significantly up-regulated in brain,gill,heart,hemolymph,and intestine after infected by WSSV.The above results indicated that PO was one of the important molecules that participate in the immune defense response in F.penicillatus.

关 键 词：长毛明对虾 酚氧化酶原 酚氧化酶 基因克隆 组织表达 

分 类 号：S917.4[农业科学—水产科学]