Lei Li-ping, Zhu Xin-ping, Yang Jing, Yang Ya-ping, Ding Li
Department of Parasitology, Capital University of Medical Sciences, Beijing 100054, China.
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2004 Oct;22(5):257-61.
To obtain the recombinant protein of an antigen gene Ts88 of Trichinella spiralis and identify the characteristics of the recombinant protein.
Ts88 cDNA obtained by immunoscreening the cDNA library of adult T. spiralis was subcloned into the pET-28c(+) expression vector and expressed in E. coli. Mice were immunized with the fusion protein incorporated into Freund' s adjuvant and the immune sera were collected. The titers of the Ts88 immune sera and the antigenicity of the recombinant protein were detected by ELISA and Western blotting. Immuno-fluorescence test was performed in order to confirm the distribution of Ts88 protein in the worm.
The fragment of Ts88 gene was expressed successfully in E. coli and a highly purified fusion protein was obtained. Immunization with the recombinant protein in mice produced high titers of antibodies, which recognized some components of native antigens of soluble proteins from adult worm of T. spiralis. Western blotting analysis showed that Ts88 recombinant antigen was recognized by all the positive sera, such as the sera from infected or immunized rabbits, from infected swine and from patients of trichinosis. Immuno-fluorescence test confirmed that Ts88 protein mainly distributed in the cuticle surface of the worm.
The Ts88 antigen gene from T. spiralis was successfully expressed. The recombinant protein presented antigenicity.
获取旋毛虫抗原基因Ts88的重组蛋白并鉴定其特性。
通过免疫筛选旋毛虫成虫cDNA文库获得的Ts88 cDNA亚克隆至pET-28c(+)表达载体并在大肠杆菌中表达。用掺入弗氏佐剂的融合蛋白免疫小鼠并收集免疫血清。采用ELISA和Western印迹法检测Ts88免疫血清的效价及重组蛋白的抗原性。进行免疫荧光试验以确定Ts88蛋白在虫体中的分布。
Ts88基因片段在大肠杆菌中成功表达，获得了高度纯化的融合蛋白。用重组蛋白免疫小鼠产生了高效价抗体，该抗体可识别旋毛虫成虫可溶性蛋白天然抗原的某些成分。Western印迹分析表明，Ts88重组抗原可被所有阳性血清识别，如来自感染或免疫兔、感染猪及旋毛虫病患者的血清。免疫荧光试验证实Ts88蛋白主要分布在虫体的角质层表面。
旋毛虫Ts88抗原基因成功表达，重组蛋白具有抗原性。
