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Identification of mitochondria epitope-related protein of Schistosoma japonicum with phage-displayed random peptide library
Author(s): 
Pages: 180-184
Year: Issue:  2
Journal: CHINESE JOURNAL OF MICROBIOLOGY AND IMMUNOLOGY

Keyword:  噬菌体肽库表位日本血吸虫线粒体融合蛋白;
Abstract: 目的筛选和鉴定重组的日本血吸虫(中国大陆株)线粒体相关蛋白rSj338的表位.方法用纯化的rSj338/26GST免疫家兔获得抗rSj338/26GST的多克隆抗体IgG,将抗体进一步纯化,获得抗rSj338单特异多克隆抗体IgG.用纯化抗rSj338抗体对噬菌体12肽库进行5轮免疫学筛选,挑取克隆;采用Western blot免疫识别,核苷酸序列测定分析其获得的表位并与rSj338/26GST进行同源性比较.将获得的不同表位的阳性克隆分别免疫小鼠,并采用Western blot及dot-ELISA方法筛选能刺激小鼠产生较高滴度抗rSj338抗体的阳性克隆.并将阳性噬菌体免疫的小鼠血清对纯化的rSj338/26GST、26GST、日本血吸虫成虫及虫卵抗原进行Western blot识别.结果经5轮免疫学筛选后挑取的32个克隆,用Western blot方法30个克隆能被抗rSj338抗体识别.核苷酸序列分析发现共有11种表位,与rSj338无一级结构的同源性.经动物免疫初步实验筛选,共获得4个免疫原性较强的阳性克隆,其免疫鼠血清均可识别rSj338/26GST、日本血吸虫成虫及虫卵抗原.结论获得了4种日本血吸虫中国大陆株线粒体相关蛋白rSj338的表位,均为模拟表位,这将为日本血吸虫病的疫苗研究开辟新的途径.
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