中华鲟B细胞活化因子的分子表征及与免疫反应相关的潜在功能  

Molecular Characterization of B Cell-Activating Factor from Chinese Sturgeon ( Acipenser Sinensis) and Its Potential Function Related to Immune Response

作　　者：夏翱文 刘榴[1] 李小花 聂东宋 廖志勇[1] XIA Ao-wen;LIU Liu;LI Xiao-hua(College of Life and Environmental Science,Wenzhou University,Wenzhou,Zhejiang 325035)

机构地区：[1]温州大学生命与环境科学学院,浙江温州325035 [2]湖南科技学院化学化工系,湖南岳阳414006

出　　处：《安徽农业科学》2021年第18期105-112,共8页Journal of Anhui Agricultural Sciences

基　　金：中国国家基础研究项目(2013CB835301)。

摘　　要：[目的]探究中华鲟B细胞活化因子(CsBAFF)的分子表征及其与免疫反应相关的潜在功能。[方法]通过免疫刺激后获得中华鲟各组织。结合生物信息学分析,构建和筛选CsBAFF的cDNA文库,并采用定量RT-PCR进行验证分析。采用分子生物学的方法进行CsBAFF的质粒构建及重组sCsBAFF的制备。采用MTT试验检测细胞的增殖,以及结合免疫印迹分析相关蛋白的表达。[结果]克隆了CsBAFF的cDNA,包含765个核苷酸的开放阅读框,编码255个氨基酸,分子量为28.9 kD。CsBAFF具有跨膜结构域、TNF结构域、furin蛋白酶裂解位点、长D-E环、1个潜在的N-连接糖基化位点和2个保守的半胱氨酸残基。CsBAFF基因主要表达于头肾和脾脏,且三价灭活疫苗和聚肌苷酸均能诱导CsBAFF基因表达。重组CsBAFF能促进中华鲟和小鼠淋巴细胞的增殖,其中保守的半胱氨酸残基Cys201和Cys214是CsBAFF发挥生理功能必不可少的位点。[结论]B细胞活化因子(BAFF)是肿瘤坏死因子(TNF)家族成员之一,在B淋巴细胞的增殖、存活、分化和成熟过程中起着重要作用。[Objective]To explore the molecular characterization of Chinese sturgeon B cell activating factor(CsBAFF)and its potential functions related to immune response.[Method]Various tissues of Chinese sturgeon were obtained after immune stimulation.Combined with bioinformatics analysis,a cDNA library of CsBAFF was constructed and screened,and quantitative RT-PCR was used for verification and analysis.Using molecular biology methods,the plasmid construction of CsBAFF and the preparation of recombinant sCsBAFF were carried out.The MTT experiment was used to detect cell proliferation,and the expression of related proteins was analyzed in combination with western blotting.[Result]In this study,the cDNA of CsBAFF was cloned.It contains an open reading frame of 765 nucleotides,encodes 255 amino acids,and has a molecular weight of 28.9 kDa.CsBAFF has a transmembrane domain,a TNF domain,a furin protease cleavage site,a long D-E loop,a potential N-linked glycosylation site and two conserved cysteine residues.CsBAFF gene is mainly expressed in head kidney and spleen,and both trivalent inactivated vaccine and polyinosinic acid can induce CsBAFF gene expression.Recombinant CsBAFF can promote the proliferation of Chinese sturgeon and mouse lymphocytes,and the conservative cysteine residues Cys201 and Cys214 are essential sites for CsBAFF to exert its physiological functions.[Conclusion]B cell activating factor(BAFF)is a member of the tumor necrosis factor(TNF)family,which plays an important role in the proliferation,survival,differentiation and maturation of B lymphocytes.

关 键 词：中华鲟 B细胞活化因子 免疫应答 

分 类 号：S917.4[农业科学—水产科学]