中华鲟IFNc基因的克隆鉴定及表达模式研究  

Cloning,identification and expression pattern of IFNc in Acipenser sinensis

作　　者：丁广义 郑楚文 张书环 杜浩 许巧情[2] 危起伟 DING Guangyi;ZHENG Chuwen;ZHANG Shuhuan;DU Hao;XU Qiaoqing;WEI Qiwei(Key Lab of Freshwater Biodiversity Conservation,Ministry of Agriculture and Rural Affairs,Yangtze River Fisheries Research Institute,Wuhan 430223,China;Institute of Chinese Sturgeon Disease,Yangtze University,Jingzhou Hubei 434024,China;Sturgeon Healthy Culture and Medicinal Value Research Center,Guizhou University of Traditional Chinese Medicine,Guiyang 550025,China)

机构地区：[1]农业农村部淡水生物多样性保护重点实验室,长江水产研究所,武汉430223 [2]长江大学中华鲟病害研究所,湖北荆州434020 [3]贵州中医药大学鲟鱼健康养殖与药用价值研究中心,贵阳550025

出　　处：《海洋渔业》2024年第5期571-578,共8页Marine Fisheries

基　　金：农业农村部淡水生物多样性保护重点实验室开放课题(LFBC1007);贵州省省级科技计划项目(黔科合基础-ZK[2022]一般507);国家自然科学基金项目(32260925)。

摘　　要：从中华鲟(Acipenser sinensis)转录组数据库中筛选和验证干扰素c(interferon c,IFNc)基因,对中华鲟IFNc基因序列进行生物信息学分析,利用qPCR研究中华鲟IFNc基因在组织中和免疫刺激条件下的表达模式。结果表明,中华鲟IFNc基因ORF序列长549 bp,编码183个氨基酸,与眼斑雀鳝(Lepisosteus oculatus)IFNc1一致性为32.13%;qPCR结果显示,中华鲟IFNc基因在各组织中呈低水平表达,在肌肉、血液、皮肤和体肾中表达量最高;中华鲟IFN-γ重组蛋白和Poly I:C诱导中华鲟肾细胞后,IFNc基因、抗粘液病毒基因(myxovirus resistance,Mx)和Viperin基因在24 h内呈显著上调。通过克隆鉴定中华鲟IFNc基因,探究了其在免疫刺激条件下的表达规律,可为中华鲟干扰素功能研究和抗病毒药物开发提供理论基础。Chinese sturgeon Acipenser sinensis interferon IFNc(AsFINc)was screened from the transcriptome database,PCR was used for cloning verification,bioinformatics was used to analyze its sequence characteristics and evolutionary status,and qPCR was used to study its expression patterns in different tissues and under different immune stimulation.Results showed that the IFNc cDNA was 549 bp in length and consisted of 183 amino acids.Bioinformatics analysis showed that A.sinensis IFNc belonged to type I interferon c subgroup.Phylogenetic tree and homology analysis showed that the IFNc of A.sinensis and Lepisosteus oculatus clustered into one branch.Gene structure analysis showed that the IFNc gene was composed of five exons and four introns,and the intron phase was zero.These differences in protein structure and physicochemical properties suggested that A.sinensis IFNc played different functions in the immune system.IFNc could be expressed in a number of tissues,including liver,blood,cheek,heart,spleen,brain,intestine,muscle,cephalic kidney,mesonephric and skin.The expression level of AsIFNc was the highest in muscle,blood and skin,indicating that AsIFNc might play an important role in muscle,blood and skin.To further understand the function of A.sinensis IFNc,kidney cells were incubated.The experimental groups were added with A.sinensis IFN-γrecombinant protein and Poly I:C,respectively,and the control group was added with the same amount of PBS.After incubation at 28℃for 1 h,8 h,24 h,the medium solution in each hole was removed,and 1 ml Trizol was added to stand at room temperature for 5 min.The solution was collected,total RNA was extracted and converted into cDNA,and then quantitative qPCR detection was performed.The experimental result showed AsIFNc could be induced by poly I:C,and was significantly upregulated with Mx and Viperin,but the upregulation ratio was not as high as Mx and Viperin,and the maximum upregulation ratio was 2.58 times.It was speculated that AsIFNc had certain antiviral function,but its specif

关 键 词：中华鲟 IFNc 抗病毒 免疫调控 

分 类 号：S917.4[农业科学—水产科学]