细粒棘球蚴(中国大陆株)抗原zw-5基因的表达、纯化及免疫学特性初步分析

目的对细粒棘球蚴(Echinococcus granulosus,Eg)抗原zw-5重组质粒进行原核表达、纯化,并初步分析重组蛋白的免疫学特性。方法从重组质粒Eg.zw-5/pGEM-T中获取抗原zw-5基因,亚克隆于表达载体pET-28a构建基因工程菌株,并表达、纯化重组蛋白;用Eg.zw-5免疫ICR小鼠,通过Western-blot、ELISA对Eg.zw-5的免疫学特性进行初步研究。结果构建原核重组表达基因工程菌株Eg.zw-5/pET-28a/BL21(DE3)plysS,并纯化出分子量为28kD的重组蛋白。West-ern-blot显示,Eg.zw-5免疫的小鼠血清能识别重组蛋白和天然抗原原头蚴中约28KD处的条带。ELISA检测显示,用Eg.zw-5免疫小鼠,可诱导产生特异性抗体IgG。结论成功表达细粒棘球蚴重组抗原Eg.zw-5,该重组蛋白有较好的免疫原性及抗原性。

Expression and purification of antigen zw-5 recombinant protein from Echiococcus granulosus (Chinese mainland strain) and the preliminary analysis on its immunogenicity

In order to exress and urify antigen zw-5 recombinant lasmid of Echinococcus granulosus（Eg） rokaryotically,as well as identify the immunogenicity of recombinant zw-5 rotein,Eg.zw-5 gene was obtained from Eg.zw-5/GEM-T recombinant lasmid and subcloned into high-level exression vector ET28a.Then the ICR mice were immunized by the urified rotein,and Western-blot and ELISA were alied to identify the immunological characteristic of the recombinant rotein.Results demonstrated that Eg.zw-5/ET28a/BL21（DE3） lysS recombinant lasmid was constructed and a 28kD recombinant rotein was obtained from urification.Western-blot showed that serum from recombinant rotein immunized mice could recognize the 28kD band of recombinant rotein and native antigen of rotoscolex.ELISA showed that the secific IgG antibody could be induced to roduct in Eg.zw-5 immunized mice and the Eg.zw-5/ET28a recombinant lasmid was constructed and exressed efficiently.These results indicate that recombiant Eg.zw-5 rotein ossesses good immunogenicity and antigenicity.