论文部分内容阅读
目的克隆表达恶性疟原虫网状细胞结合蛋白同源体5(PfRh5)F2片段,并评价其抗原性。方法 PCR扩增目的基因片段,克隆到表达载体pTGub21b中,构建PfRh5F2/pTGub21b原核表达载体。IPTG诱导表达目的基因,SDS-PAGE电泳分析表达产物,并用Western blot检测其抗原性。结果成功构建了PfRh5F2/pTGub21b原核表达系统,并在大肠杆菌中可溶性高效表达,表达产物能被恶性疟患者血清识别,而重组抗原免疫鼠血清也能识别恶性疟患者血样中的相应抗原。结论恶性疟原虫PfRh5 F2片段在大肠杆菌中获得高效表达且表达产物具有良好的抗原性和免疫原性。
Objective To clone and express the F2 fragment of Plasmodium falciparum reticulocyte binding protein 5 (PfRh5) and to evaluate its antigenicity. Methods The target gene fragment was amplified by PCR and cloned into the expression vector pTGub21b to construct the prokaryotic expression vector PfRh5F2 / pTGub21b. IPTG induced expression of the target gene, SDS-PAGE electrophoresis analysis of expression products, and Western blot detection of antigenicity. Results The prokaryotic expression system of PfRh5F2 / pTGub21b was successfully constructed and expressed in E.coli. The expressed product was recognized by the serum of falciparum malaria patients, while the serum of recombinant murine immunized mice could recognize the corresponding antigens in blood samples of patients with falciparum malaria. Conclusion The P. falciparum PfRh5 F2 fragment is highly expressed in E. coli and the expressed product has good antigenicity and immunogenicity.