Suppr超能文献

[恶性疟原虫乳酸脱氢酶的表达及免疫活性特征]

[Expression and immunocompetence characterization of Plasmodium falciparum lactate dehydrogenase].

作者信息

Wu Y S, Li M, Dong W Q, Li Y J

机构信息

Institute of Tropical Medicine, First Military Medical University, Guangzhou 510515.

出版信息

Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2001;19(2):80-3.

Abstract

OBJECTIVE

To express lactate dehydrogenase (LDH) gene of Plasmodium falciparum FCC1/HN in the E. coli TG1 and analyse its immunocompetence.

METHODS

The LDH gene of the P. falciparum was specifically amplified by polymerase chain reaction, and the recovered gene fragment was cloned into pGEX-4T-1 vector for expression of fusion protein with glutathione S-transferase(GST). The recombinant plasmid was transformed into the E. coli TG1. Four mice (Kunming strain) were immunized with purified expressed protein(antigen) and the polyclonal antibodies were collected. The immunocompetence of recombinant protein was analysed by ELISA and Western-blot.

RESULTS

The LDH gene of P. falciparum was successfully expressed in the E. coli TG1. The expressed protein exhibited a specific reaction with immune sera obtained from rabbits immunized with P. falciparum. The specific humoral responses were induced in mice and the titer of the specific antibody was 1:16 by two-dimensional diffusion assay.

CONCLUSION

The LDH gene of P. falciparum has been successfully expressed in the E. coli TG1 and the expressed protein has high antigencity.

摘要

目的

在大肠杆菌TG1中表达恶性疟原虫FCC1/HN乳酸脱氢酶(LDH)基因并分析其免疫活性。

方法

采用聚合酶链反应特异性扩增恶性疟原虫的LDH基因,回收基因片段并克隆至pGEX-4T-1载体,用于与谷胱甘肽S-转移酶(GST)融合蛋白的表达。将重组质粒转化至大肠杆菌TG1。用纯化的表达蛋白(抗原)免疫4只小鼠(昆明种),收集多克隆抗体。通过酶联免疫吸附测定(ELISA)和蛋白质印迹法分析重组蛋白的免疫活性。

结果

恶性疟原虫的LDH基因在大肠杆菌TG1中成功表达。表达的蛋白与用恶性疟原虫免疫的兔获得的免疫血清呈现特异性反应。在小鼠中诱导了特异性体液反应,二维扩散试验显示特异性抗体效价为1:16。

结论

恶性疟原虫的LDH基因已在大肠杆菌TG1中成功表达,表达的蛋白具有高抗原性。

文献检索

告别复杂PubMed语法,用中文像聊天一样搜索,搜遍4000万医学文献。AI智能推荐,让科研检索更轻松。

立即免费搜索

文件翻译

保留排版,准确专业,支持PDF/Word/PPT等文件格式,支持 12+语言互译。

免费翻译文档

深度研究

AI帮你快速写综述,25分钟生成高质量综述,智能提取关键信息,辅助科研写作。

立即免费体验