Wu Y S, Li M, Dong W Q, Li Y J
Institute of Tropical Medicine, First Military Medical University, Guangzhou 510515.
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2001;19(2):80-3.
To express lactate dehydrogenase (LDH) gene of Plasmodium falciparum FCC1/HN in the E. coli TG1 and analyse its immunocompetence.
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.
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.
The LDH gene of P. falciparum has been successfully expressed in the E. coli TG1 and the expressed protein has high antigencity.
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。
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。
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。
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。
