南方农业学报2023，Vol.54Issue(11)：3405-3415,11.DOI:10.3969/j.issn.2095-1191.2023.11.027
乌鳢半胱氨酸蛋白酶3基因克隆及表达分析
Cloning and expression analysis of cysteinyl aspartate specific proteinase 3(caspase-3)gene in Channa argus
摘要
Abstract
[Objective]Cloning and expression analysis of cysteinyl aspartate specific proteinase-3 gene(caspase-3 gene)of Channa argus were conducted,which provided a theoretical basis for exploring the role of this gene in the im-mune response of C.argus and its role in the immunity of other fishes.[Method]According to the genome information of C.argus published by GenBank,the caspase-3 gene sequence was cloned and the deduced caspase-3 gene sequence bioin-formatics analysis was performed by real-time fluorescence quantitative PCR(qRT-PCR)to detect the expression distribu-tion of caspase-3 gene in various tissues of healthy C.argus and the temporal expression under different stimuli(Seriola quinqueradiata Nocardia seriolae,LPS and PolyI:C).[Result]The caspase-3 gene sequence was 855 bp and encoded 284 amino acid residues,with a protein molecular mass of 31.069 kD and a theoretical isoelectric point of 6.17.The cas-pase-3 protein contained 1 CASc domain.Multiple sequence alignment showed that the amino acid sequence of caspase-3 of C.argus and the caspase-3 of Lutjanus peru,reaching 81.7%.Phylogenetic analysis showed that caspase-3 of C.argus and caspase-3 of L.peru clustered into a single lineage.The results of qRT-PCR showed that,the caspase-3 gene of C.ar-gus in the tested tissue(muscle,skin,gills,heart,thymus,brain,head kidney,spleen,liver and blood)were ex-pressed.The spleen and head kidney had the highest expression.Upon stimulation with the different stimuli,the expres-sion level of caspase-3 gene in the spleen,head kidney,blood and liver tissues of C.argus all changed significantly(P<0.05);After incubation of head kidney leukocyte with S.quinqueradiata N.seriolae,LPS and PolyI:C,the caspase-3 gene expression was all significantly increased,and it had an obvious time-dependence.[Conclusion]The caspase-3 gene of C.argus may play an important role in the immune response against bacterial and viral invasion.关键词
乌鳢/半胱氨酸蛋白酶3基因(caspase-3基因)/组织分布/免疫应答Key words
Channa argus/cysteinyl aspartate specific proteinase 3 gene(caspase-3 gene)/tissue expression/im-mune response分类
农业科技引用本文复制引用
赵浩航,丘金珠,周秀莹,唐怀庆,赵天珍,梁富,宋海霞,喻大鹏,宋长江..乌鳢半胱氨酸蛋白酶3基因克隆及表达分析[J].南方农业学报,2023,54(11):3405-3415,11.基金项目
国家重点研发计划项目(2020YFD0900201) （2020YFD0900201）
中山市第二批社会公益与基础研究项目(2021B2058) National Key Research and Development Program of China(2020YFD0900201) （2021B2058）
The second Batch of Social Welfare and Basic Research Projects in Zhongshan(2021B2058) （2021B2058）
