恶性疟原虫网状细胞结合蛋白同源体5F1段的克隆、表达及初步鉴定.pdfVIP

624 中同人兽共患病学报 2010. 26 (7) Chinese Journal of Zoonoses 文章编号: 1002-2694(2010 )07- 0624 - 03 恶性症原虫网状细胞结合蛋白同源体5 Fl 段的克隆、表达及初步鉴忘赞 郝文泼，李宏，邓小英，廖小青，李 明，罗树红 摘 要:目的 克隆表达恶性疙原虫网状细胞结合蛋白同源体 5CPfRh5)Fl 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 pET28a( +)中，构建 PfRh5Fl/pET28a 原核表达载体。 IPTG 诱导表达目的基因 .SDS 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 PfRh5Fl / pET28a 原核表达系统.并在大肠杆菌中 以包涵体形式高效表达，表达产物能被恶性芜原虫感染患者血清识别，而不能被问日芜原虫感染患者及正常人血清识别。结 论 恶性芜原虫 PfRh5 F1 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。 关键词:恶性在原虫 g 网状细胞结合蛋白同源体 5; 克隆 z 表达 中图分类号:R382.3 文献标识码:A Cloning , expression and identification of reticulocyte-binding protein homologue 5 Fl fragment in Plasmodium falciparum HAO Wen-bo ,Ll Hong ,DENG Xiao-ying ,LlAO Xiao-qing ,Ll Ming.LUO Shu-hong (Institute of Antibody Engineering. Schοο1 of Biotechnology. Southern Medical University. Guangzhou 510515. China) ABSTRACT:ln order to clone and express reticulocyt e- binding protein homologue 5 CPfRh5) F1 fragment gene of Plasmo- dium falci parum. the 429bp PfRh5 gene C91-519) was specifically amplified by polymerase chain reaction and cloned into pET28aC +) vector. The recombinant plasmid was then transformed and induced to express in E. coli Rosetta. The expressed product was analyzed by SD5and Western Blot respectively. And the expressed protein was insoluble with a size of a- bout 2 1. 9 kDa as predicted. Also. it exhibited a specific reaction with immune sera obtained from patients with Pf maleria. These results demonstrate that the PfRh5 Fl fragment has been successfully expressed and the expressed protein has certain an- t1 gemclty. KEY WORDS: Plasmodium falciparum; reticulocyt e-binding protein homologue 5; clone; expression