斑点叉尾鮰血浆铜蓝蛋白的全长cDNA克隆及表达特征分析

Cloning and expression analysis of ceruloplasmin cDNA in channel catfish Ictalurus punctatus

中文关键词: 斑点叉尾鮰  血浆铜蓝蛋白  感染  差异表达

英文关键词: Ictalurus punctatus  Ceruloplasmin  Infection  Differential expression

中文摘要:

      本研究克隆了斑点叉尾鮰中血浆铜蓝蛋白基因全长cDNA序列，其中5′非翻译区(5′-UTR)25 bp、3′非翻译区(3′-UTR) 860 bp、开放阅读框(Open reading frame，ORF) 3 225bp，编码一条含有1 074个氨基酸的多肽链。与其他真核生物血浆铜蓝蛋白成员进行同源性比较，表现出较高的保守性。本研究利用荧光实时定量PCR技术分析了斑点叉尾鮰血浆铜蓝蛋白基因在正常组织和被不同病菌感染后的肝脏、肠、头肾及脾脏中的表达。结果发现，正常组织中血浆铜蓝蛋白基因在正常肝脏中表达最强烈。4种病原（迟钝爱德华氏菌、海豚链球菌、嗜水气单胞菌和斑点叉尾鮰呼肠孤病毒）引起的血浆铜蓝蛋白基因的应答在不同组织中都有差异。血浆铜蓝蛋白基因对海豚链球菌的应答最为显著，而对呼肠孤病毒的应答最不明显。而且不难发现，感染4种病原菌后，血浆铜蓝蛋白基因在头肾中的表达都出现明显的上调。

英文摘要:

      Ceruloplasmin is a ferroxidase combining more than 95% copper atoms in plasma that plays roles in copper transport and regulates iron homeostasis. In this study, a full-length cDNA of ceruloplasmin was cloned from channel catfish Ictalurus punctatus. The cDNA of ceruloplasmin has a length of 4 110 bp, composed of a 25 bp 5′ UTR, an 860 bp 3′ UTR and a 3 225 bp ORF, encoding a polypeptide of 1,074 amino acids. Alignments showed that catfish ceruloplasmin was conserved among others. Quantitative real-time PCR was conducted to detect expression profiles of ceruloplasmin in normal tissues and four tissues (liver, headkidney, intestine and spleen) after infection by different pathogens: Edwardsiella tarda, Streptococcus iniae, Aeromonas hydrophila, and channel catfish Hemorrhage Reovirus (CCRV). The expression profile in liver was the highest in normal tissues. S. iniae induced the expression of ceruloplasmin to the highest extent, and CCRV virus to the lowest extent. These results provide a basis for further research aimed at exploring the precise immune-related molecular mechanism of ceruloplasmin in channel catfish.