恶性疟原虫杂合抗原基因的表达及其产物免疫功能的研究  

A Study on the Expression of Human P. Falciparum Hybrid Poly-petide Antigen Gene and the Immunological Function of the Expressed Products

作　　者：王昌才[1] 钟雄林[1] 陈仕荣[1] 胡亚芳[1] 马维芳[1] 王启松[2] 闵永洁 

机构地区：[1]第一军医大学分子生物学研究所,广州510515 [2]复旦大学遗传学研究所,上海200433

出　　处：《上海免疫学杂志》1997年第3期139-143,共5页Shanghai Journal of Immunology

基　　金：军队重点资助项目

摘　　要：化学合成的人恶性疟原虫杂合多肽抗原 AB 基因,编码子孢子 CSP重复序列 NANP、CSPTh2R和红内期spf35.1、spf83.1、spf55.1、spf83.18、pf155/RESA-5’端重复区共7个不同免疫功能表位,与大肠杆菌质粒pWR450-1重组,构建成pWR450-1/AB表达载体。在乳糖操纵子调控下以β-半乳糖苷酶-恶性疟原虫杂合多肽抗原融合蛋白形式,在大肠杆菌中高效表达。表达产物经SDS-PAGE电泳分离纯化后,加福氏佐剂免疫家兔,诱导产生较高滴度的抗血清。抗血清对人恶性疟原虫体外生长有明显的抑制作用,24h抑制率为65.92％;72h的抑制率为72.63％。对照血清无抑制作用。分离纯化的表达产物能与鼠抗恶性疟原虫和疟疾患者抗血清起免疫反应。这些结果表明合成基因的表达产物具有较好的免疫原性。A human P.falciparum hybrid polypeptide antigen gene encoding seven different immudominant epitopes of sporozoitic stage (ig. CSP repeat NANP and CSP TH2R) and asexsual crythroeyte stage (ig. 8pf35.1,spf83.1,Spf55.1,spf83.18 and spf 155 /RESA-5' repeat regions) have been recombined with the expression plasmid pwR450-1 and transformed into E.coli JM103. Under the Lac promoter control, the hybrid fusion gene was expressed at high level in E.coli and the expression products were a fusion protein of p-galactosidase-hybrid polypeptide antigen.The expression products were purified by SDS-PAGE preparation electrophoresis. The purified products were used to immunize rabbits with Freund's adjuvant and induced high titres of anti-serum to this products. The antiserum could not only specially react to the fussion protein, but also obviously inhibit the growth of P. falciparum in vitro. The inhibition rate after 24 hours was 65.9% and that after 72 hours was 72.9%. The purified products also could react wfnrthe mouse antiserum against P. falciparum and the sera of malaria patients. These results indicate that the expression products of the synthetic hybrid gene have a good immunogenicity against P. falciparum antigen.

关 键 词：恶性疟原虫 基因重组表达 抗原 

分 类 号：R531.303[医药卫生—内科学]