摘要
目的克隆表达恶性疟原虫网状细胞结合蛋白同源体5(PfRh5)F1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PfRh5F1/pET28a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WesternBlot检测其抗原性。结果成功构建了PfRh5F1/pET28a原核表达系统,并在大肠杆菌中以包涵体形式高效表达,表达产物能被恶性疟原虫感染患者血清识别,而不能被间日疟原虫感染患者及正常人血清识别。结论恶性疟原虫PfRh5F1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。
In order to clone and express reticulocyte-binding protein homologue 5（PfRh5）F1 fragment gene of Plasmodium falciparum,the 429bp PfRh5 gene（91-519）was specifically amplified by polymerase chain reaction and cloned into pET28a（＋）vector.The recombinant plasmid was then transformed and induced to express in E.coli Rosetta.The expressed product was analyzed by SDS-PAGE and Western Blot respectively.And the expressed protein was insoluble with a size of about 21.9 kDa as predicted.Also,it exhibited a specific reaction with immune sera obtained from patients with Pf maleria.These results demonstrate that the PfRh5 F1 fragment has been successfully expressed and the expressed protein has certain antigenicity.
出处
《中国人兽共患病学报》
CAS
CSCD
北大核心
2010年第7期624-626,共3页
Chinese Journal of Zoonoses
基金
国家重点基础研究发展计划973专项(No.2007CB513101)
国家自然科学基金(No.30200238)联合资助
关键词
恶性疟原虫
网状细胞结合蛋白同源体5
克隆
表达
Plasmodium falciparum
reticulocyte-binding protein homologue 5
clone
expression
