摘要
目的克隆并表达恶性疟原虫重组复合抗原，为疟疾疫苗研究奠定基础。方法将恶性疟原虫MSP1中类表皮生长因子1（EGF—1）基因与人工化学合成的抗原复合基因PfCMR串接，插入高效非融合型蛋白表达载体PBV220，转化大肠杆菌DH5a．转化菌经42℃热诱导表达，表达产物用Westernblot和Dot—ELISA分析。结果构建成功重组质粒PBV220／PfCMR—EGF─1，经热诱导后表达出合外源基因产物的非融合蛋白质，分子量为16．5kDa，表达产物与恶性疟原虫抗原免疫鼠血清产生特异免疫反应。工程菌连续传代，未见重组质粒丢失，表达效率无明显改变。结论用表达载体pBV220表达出的恶性疟原虫重组复合抗原具有一定的免疫活性。该重组质粒可在宿主菌DH5a内长期稳定存在。
Aim Cloning and expressing the protective epitopes of Plasmodium falciparum antigens as malaria vaccine. Method The first EGF--like domain of MSPI and the chemically synthesized hybrid gene Pf CMR were recombined and inserted into expression vector pBV220,then the recombinant vector was transferred into E. colt DH5a. The genetically engineered bacteria shaked at 30>C and expressed at 42C. The expressed protein (non-fusion prolein )was isolated and analyzed by SDS--PAGE .Western blot and act -ELISA' Results Successfully constructing recombint plasmid pBV220/R/tMR--EGF-- 1. And after heating at 42oC .the bacteria harboring recombinant plasmid expressed the non fusion protein of exogenous gene products,its molecular weights was about 16. 5kDa. The non-fusion protein could strongly react with mouse anti -Plasmodium falciParum serum by Dot-- ELISA and Western blot. After 34 generations of continuous culture,we found that the recombinant plasmid was stable in DH5a and did not have obvious toxicity' Conclusion The non--fusion protein harbored the antigenic epitopes of Plasmodium jalciparum,it has brought the possibility of malaria vaccine a step further.
出处
《中国人兽共患病杂志》
CSCD
北大核心
1998年第3期9-12,共4页
Chinese Journal of Zoonoses
关键词
恶性
疟原虫
复合基因
非融合蛋白
疟疾疫苗
Plasmodium falciparum Hybrid gene Non-fusion protein Expression Malaria vaccine
