【摘要】 目的 研究百日咳毒素S1(Pertussis toxin s1 subunit)亚单位在大肠杆菌中的表达及其免疫学特性。方法 采用PCR法从百日咳杆菌染色体中获得S1亚单位的结构基因并克隆到质粒pCR2.1-TOPO中,转化宿主菌DH5α和TOPO10。用抗S1的单抗1B7检测表达产物。用粗制的重组S1(rS1)免疫小鼠,做主动免疫保护实验。rS1免疫家兔,检测rS1的免疫原性。并以家兔免疫血清在小鼠进行被动免疫保护实验。结果 rS1可在大肠杆菌中稳定高效表达,表达量分别占细胞总蛋白量的14％(DH5α)和24.2％(TOPO10)。rS1能被1B7识别,在小鼠主动免疫实验中,保护率可达56.3％。家兔免疫血清ELISA效价为1:32 000。抗rS1血清对小鼠被动的免疫保护率为100％。结论 rS1具有良好免疫原性,为百日咳基因工程疫苗或亚单位疫苗研制及百日咳免疫机理研究提供了依据。更多还原

【Abstract】 Objective To express pertussis toxin S1 subunit in E. coli and study its immunogenicity. Methods The structural gene encoding pertussis toxin SI subunit of Bordetella pertussis was amplified by PCR, cloned into vector pCR2.1 - TOPO, and transformed to E. coli strain DH5α and TOP010 respectively. The expressed product was detected by anti - S1 McAb 1B7. The active immunity of crude rS1 was tested in mice by intraceiebral challenge, and the immunogenicity of it was tested in rabbits. The immune rabbit sera were used for passive immunity test in mice. Results rS1 was stably and highly expressed in E. coli. The expressed product in DH5α and in TOP010 contained 14% and 24.2% of total cellular protein respectively. The rSl was recognized by McAb 1B7, and the protective rate of it in acitive immunity test was 56.3% . The ELISA titer of rabbit serum immunized with rSl was 1:32000, and the protective rate of it in passive immunity test was 100% .Conclusion The expressed rSl showed good immunogenicity. The study provided a basis for the preparation of recombinant pertussis or subunit vaccine, as well as research of immune mechanism of pertussis .更多还原