脑膜炎奈瑟氏菌NspA基因的克隆及序列分析

摘要

为了获得脑膜炎奈瑟氏菌(Nesseria meningitidis,Nm)表面蛋白A(NspA)基因,从脑膜炎患者的血液中分离出脑膜炎奈瑟氏菌并对其进行鉴定.然后根据NspA基因的核苷酸序列,设计合成一对特异性引物,通过聚合酶链式反应(PCR)技术体外扩增得到NspA基因.进行克隆、测序及序列分析.结果显示,其核苷酸序列同源性与Genbank报道的标准菌株相比同源性为100%.克隆获得的基因保持了脑膜炎奈瑟氏菌表面蛋白NspA基因的完整性,保留了其抗原性,为以后进一步开发脑膜炎基因工程疫苗奠定了实验基础.%For amplification ofthe Neisseria meningitis surface protein A gene (NspA) , Nesseriameningitidiswasisolated and identified from the blood of patient with meningitis. Using the genome DNA as template, NapA gene was amplified by PCR technique, sub-cloned into pMD18-T vector, and the ligation product were transformed into E.coli DH5α. After purification of the recombinant plasmid, and identification using PCR and restriction enzyme, the interest gene was identified by sequence analysis. The results showed that the nucleotide sequence homology with the standard strains reported was 100％. Theinterest gene, NspA, retained its antigenicity, providing the foundation for the development of genetic engineering vaccine against Neisseria Meningitis.