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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。
Chemical synthesis of human P. falciparum hybrid polypeptide antigen AB gene, encoding sporozoite CSP repeats NANP, CSPTh2R and red-phase spf35.1, spf83.1, spf55.1, spf83.18, pf155 / RESA-5 ’ A total of 7 different immunofunctional epitopes in the repeat region were recombined with E. coli plasmid pWR450-1 to construct pWR450-1 / AB expression vector. It is highly expressed in Escherichia coli under the control of lactose operon in the form of β-galactosidase-Plasmodium falciparum hybrid polypeptide antigen fusion protein. After the expressed product was purified by SDS-PAGE electrophoresis, rabbits were immunized with Fortune’s adjuvant to induce higher titers of antiserum. Antiserum inhibited the growth of human P. falciparum in vitro with the inhibitory rate of 65.92% at 24 h and 72.63% at 72 h. Control serum no inhibitory effect. The isolated and purified expression product can react immunologically with the antisera of the anti-Plasmodium falciparum and malaria patients. These results show that the synthetic gene expression product has better immunogenicity.