微小隐孢子虫TSP3蛋白的定位分析  

Localization of Cryptosporidium parvum TSP3 Protein

作　　者：王科杰[1] 邵清延 张天宇[1] 王东强[1] 尹继刚[1] 

机构地区：[1]吉林大学人兽共患病研究所,人兽共患病研究教育部重点实验室,长春130062

出　　处：《吉林农业大学学报》2017年第2期199-203,共5页Journal of Jilin Agricultural University

基　　金：国家自然科学基金项目(31272549)

摘　　要：通过对微小隐孢子虫TSP3氨基酸序列进行生物信息学分析设计多肽片段,偶联KLH载体蛋白免疫家兔制备多克隆抗体,采用间接ELISA方法检测血清效价并利用特异性亲和层析的方法纯化抗体。以该抗体作为一抗,利用Western blot检测虫体天然蛋白质在体内的表达情况,通过间接免疫荧光方法进行亚细胞定位分析。结果表明:制备的多克隆抗体能够特异性识别虫体天然蛋白,亚细胞定位发现其位于子孢子以及裂殖子的顶端。Bioinformatics analysis of the Cryptosporidium parvum TSP3 amino acid sequence was conducted to design peptide fragments.Polyclonal antibody was obtained from rabbits immunnized with KLH-Peptide.Indirect ELISA was adopted to detect serum titer and antibody was purified by specific affinity chromatography.Polyclonal antibody was used as primary antibody to detect the expression of TSP3 by Western blot and subcellular localization by indirect immunofluorescence assay.The results showed that the prepared polyclonal antibody could specifically recognize the native protein of the parasite,and the subcellular localization was found to be on the top of sporozoites and merozoites.The study has laid foundation for the follow-up researches on the protein.

关 键 词：微小隐孢子虫 多肽 TSP3 多克隆抗体 

分 类 号：S855.99[农业科学—临床兽医学]