黄河鲤CFH基因的克隆、组织表达及对病原类似物的表达响应  

Molecular Cloning,Expression in Some Tissues and Expression Response to Pathogenic Analogue Stimulation of CFH Gene in Yellow River Common Carp Cyprinus carpio

作　　者：赵彤 朱雷 高园园 孔祥会 ZHAO Tong;ZHU Lei;GAO Yuanyuan;KONG Xianghui(Engineering Laboratory of Henan Province for Aquatic Animal Disease Control,College of Fisheries,Henan Normal University,Xinxiang 453007,China)

机构地区：[1]河南师范大学水产学院,河南新乡453007

出　　处：《水产学杂志》2023年第2期35-42,共8页Chinese Journal of Fisheries

基　　金：河南省自然科学基金(212300410177);河南省重点科技攻关项目(212102110107)。

摘　　要：补体调节因子H(Complement factor H,CFH)为一类重要的补体调节蛋白,在调节补体激活反应和抵御病原感染过程中发挥重要作用。为研究黄河鲤(Cyprinus carpio YR)CFH基因在病原感染中的作用,本文克隆获得了体质量(20±2)g黄河鲤CFH基因开放阅读框序列(命名为CcCFH),进行生物信息学分析后,用荧光定量PCR(RT-PCR)技术分析其在肝脏、脾脏、头肾、鳃、心脏、皮肤、肠和体肾等组织中的表达水平以及腹腔注射100μL浓度为0.2 mg/mL的LPS溶液、100μL浓度为0.2 mg/mL的Poly I:C溶液和100μL生理盐水(对照组)后3 h、6 h、12 h、24 h、48 h和72 h脾脏、头肾和体肾等组织的表达变化。结果显示:CcCFH基因开放阅读框全长2 817 bp,编码939个氨基酸,包含一个信号肽和15个CCP重复结构域。同源性比对和系统进化分析显示:CcCFH基因与鲤科鱼类相似性最高。组织分布结果表明:CcCFH基因广泛分布在健康黄河鲤各组织器官中,在肝脏和皮肤中表达量最高。LPS和Poly I:C刺激后,CcCFH基因在脾脏中呈现先上升后下降的表达趋势,在刺激后的24 h出现表达峰值;在头肾中呈逐渐上升的表达趋势,在72 h表达水平最高;在体肾中3 h表达水平最高,之后呈现逐渐下降的表达趋势。以上研究结果表明,CcCFH基因参与了黄河鲤对病原的响应过程,可能在鱼类抗病原感染的免疫响应中发挥重要作用。As an important complement regulatory protein,complement factor H(CFH)plays a crucial role in regulation of the complement activation response and resisting pathogen infection.In order to investigate the immune role of CFH gene in Yellow River common carp Cyprinus carpio YR(known as CcCFH),the open reading frame sequence of CcCFH was cloned.Then the bioinformatics analysis of CcCFH was carried out,the expression level of CcCFH in different tissues including liver,spleen,head kidney,gill,heart,skin,intestine and body kidney,and the expression change profiles of CcCFH were detected in spleen,head kidney and body kidney of YR intraperitoneally injected with 100μL of LPS solution at a concentration of 0.2 mg/mL,100μL of Poly I:C at a concentration of 0.2 mg/mL and 100μL of normal saline(control group)at 3 h,6 h,12 h,24 h,48 h and 72 h after injection by fluorescence quantitative PCR(RT-PCR)technology.The results showed that the open reading frame(ORF)of CcCFH was found to be 2,817 bp encoding a deduced protein with 939 amino acids.The amino acid sequence homology and phylogenetic analysis revealed that CcCFH had a high similarity with cyprinid fishes.CcCFH was ubiquitously expressed in different tissues in YR,with the maximal expression in liver.The expression levels of CcCFH were significantly changed in spleen,headkidney and kidney after LPS and Poly I:C infection(P<0.05).The findings suggest that the CcCFH may play an important role in pathogen recognition and immune response.

关 键 词：黄河鲤 CFH 基因克隆 病原类似物 免疫响应 

分 类 号：S968.22[农业科学—水产养殖]