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本研究克隆了斑点叉尾中血浆铜蓝蛋白基因全长cDNA序列,其中5′非翻译区(5′-UTR)25bp、3′非翻译区(3′-UTR)860bp、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。
In this study, we cloned the full-length cDNA sequence of ceruloplasmin in the channel catfish, including 25bp 5 ’untranslated region (5’-UTR) and 860bp 3’-untranslated region (3’-UTR) Open reading frame (ORF) 3 225 bp, encoding a polypeptide chain of 1 074 amino acids. Homology comparison with other ceruloplasmin ceruloplasmin members showed high conservation. In this study, Fluorescence real-time quantitative PCR was used to analyze the expression of ceruloplasmin in the liver, intestine, head kidney and spleen of normal catfish and different pathogenic bacteria. The results showed that the normal tissue ceruloplasmin gene expression in the normal liver in the most intense. The response of the four ceruloplasmin genes (Edwardsiella flexneri, S. iniae, Aeromonas hydrophila, and Channel catfish reovirus) caused differences in the plasma ceruloplasmin genes in different tissues. The response of ceruloplasmin to streptococci was the most significant, while the response to reovirus was the least pronounced. And not difficult to find, infected with four kinds of pathogens, ceruloplasmin gene expression in the head kidney were significantly increased.