用NcoⅠ和KpnⅠ双酶切克隆质粒pMD-18T-NspA及可以在乳酸菌与大肠杆菌之间穿梭表达的载体质粒pW425et,将NspA基因与pW425et表达载体用T4DNA连接酶相连,构建出重组质粒pW425et-NspA,将其转入至E.coliDH5α感受态细胞中,筛选阳性重组子进行SDS-PAGE和Western-blot检测,经蛋白电泳可见约18 kD的融合蛋白,Western-blot分析表明该蛋白具有与脑膜炎奈瑟氏菌抗体的反应原性。 The recombinant plasmid pW425et-NspA was constructed by digesting the cloned plasmid pMD-18T-NspA with NcoI and KpnI and the shuttle vector pW425et, which can shuttle between Lactobacillus and E.coli. The NspA gene and pW425et expression vector were ligated with T4 DNA ligase to construct the recombinant plasmid pW425et-NspA , Which was then transferred into E. coli DH5α competent cells. The positive recombinant was screened by SDS-PAGE and Western-blot. The fusion protein was about 18 kD by protein electrophoresis. Western-blot analysis showed that the recombinant protein has the same immunogenicity as meningitis Neisserial antibody.