中华鳖趋化因子CXCL10的分子特征及其对细菌感染的响应  

Molecular characterization of CXCL10 in Chinese soft-shelled turtle(Pelodiscus sinensis)and its response to bacterial infection

作　　者：张爱茹 黄秋雅 吴彬 钱国英[1] 张海琪 徐洁皓 ZHANG Airu;HUANG Qiuya;WU Bin;QIAN Guoying;ZHANG Haiqi;XU Jiehao(College of Biological and Environmental Sciences,Zhejiang Wanli University,Ningbo 315100,China;Key Laboratory of Healthy Freshwater Aquaculture,Ministry of Agriculture and Rural Affairs,Key Laboratory of Fish Health and Nutrition of Zhejiang Province,Zhejiang Institute of Freshwater Fisheries,Huzhou 313001,China)

机构地区：[1]浙江万里学院生物与环境学院,浙江宁波315100 [2]浙江省淡水水产研究所,农业农村部淡水渔业健康养殖重点实验室,浙江省鱼类健康与营养重点实验室,浙江湖州313001

出　　处：《水产学报》2024年第6期169-178,共10页Journal of Fisheries of China

基　　金：浙江省淡水水产研究所,农业农村部淡水渔业健康养殖重点实验室开放基金(ZJK202011);浙江省生物工程一流学科(A类)自设课题项目(CX2020030);浙江省大学生科技创新活动计划(新苗人才计划)(2020R419012);浙江省大学生创新创业训练计划项目(S202010876083)。

摘　　要：为研究趋化因子基因cxcl10的表达特征及其对外源病原微生物的响应,实验以中华鳖为对象,利用生物信息学分析CXCL10蛋白的分子特征及系统进化,通过荧光定量PCR(qPCR)技术检测该基因在健康中华鳖各组织中的表达分布及其在革兰氏阳性菌金黄色葡萄球菌和革兰氏阴性菌嗜水气单胞菌胁迫下mRNA的表达规律。结果显示,该蛋白分子质量为10.89 ku,等电点为9.62,存在信号肽和磷酸化位点,单链包含1个N端的loop区、1个α螺旋和3个反向平行β折叠;该基因在健康中华鳖的肝脏和心脏中表达量最高,在血细胞中表达量最低;两种细菌的刺激能促使中华鳖组织器官中cxcl10及炎症相关因子基因tnf-ɑ和il-1βmRNA表达量的显著上调。研究表明,cxcl10可能在中华鳖机体抗菌免疫过程中发挥重要作用。本实验可为中华鳖病害防治提供理论基础。Chemokines are small molecular secreted proteins with chemoattractive properties.They have bactericidal activity and the ability to recruit leukocytes and activate immune cells.CXCL10 is a kind of CXC chemokine with specificity for CXCR3 receptors,which has been shown to mediate the generation of Th1-type immune responses and have antibactericidal effects similar to defensins.Therefore,CXCL10 might be used as an alternative to antibiotics for bacterial diseases in aquaculture industry.In this study,Pelodiscus sinensis was used to study the expression characteristics of cxcl10 and its response to gram-positive bacteria(Staphylococcus aureus)and gram-negative bacteria(Aeromonas hydrophila).The molecular characteristics and systematic evolution of CXCL10 protein in P.sinensis were analyzed by bioinformatics software.The expression and distribution of cxcl10 mRNA in healthy P.sinensis tissues and that after S.aureus or A.hydrophila challenge were detected by quantitative PCR(qPCR).The results showed that the open reading frame of CXCL10 was 303 bp,encoding a protein of 100 amino acids.The molecular weight was 10.89 ku and the estimated isoelectric point was 9.62.P.sinensis cxcl10 had 8 phosphorylation modification sites and a signal peptide.Each single strand of P.sinensis CXCL10 contains one N-terminal loop region,oneαhelix and three reverse parallelβ-folds.Phylogenetic analysis showed a close relationship between P.sinensis CXCL10 with avian and reptile CXCL10s.P.sinensis cxcl10 basal levels were higher in the liver and heart than that in other organs of health turtles.The stimulation of A.hydrophila and S.aureus led to significant upregulation of P.sinensis cxcl10 and inflammation-related factors tnf-ɑand il-1βin liver,spleen and blood cells.These results suggested that P.sinensis CXCL10 had a similar protein molecular structure with the cationic antimicrobial peptide.P.sinensis cxcl10 was widely expressed in various tissues of P.sinensis,and up-regulated in response to the stimulation of gram-negative bacteri

关 键 词：中华鳖 CXCL10 MRNA表达 生物信息学 

分 类 号：Q786[生物学—分子生物学] S947.1[农业科学—水产养殖]