摘要
[目的]研究淋病奈瑟菌标准株(NG 29403和NG 29400)外膜蛋白NspA基因和NspA融合蛋白结构特点,为淋病奈瑟菌疫苗靶蛋白的选择提供依据。[方法]PCR扩增淋病奈瑟菌标准株NG 29400和NG 29403 NspA基因,与表达质粒pET-30c(g+)构建重组子NspA-pET-30c(+),阳性克隆子经双酶切鉴定后进行基因测序。以生物软件对NspA基因进行生物信息分析,并在线预测其蛋白质结构。[结果]成功构建NspA-pET-30c(+)重组质粒,双酶切获得0.5kb和5.4kb预期产物。基因序列分析表明淋病奈瑟菌NspA基因有较高同源性,两标准株与GenBank已有序列均有98%同源性,且与脑膜炎奈瑟菌NspA基因高度近似。蛋白预测显示NspA融合蛋白有40%以上为环状结构,可能为主要功能区。三级结构与脑膜炎奈瑟菌NspA相似。[结论]淋病奈瑟菌NspA基因具有高度保守性,与脑膜炎奈瑟菌NspA基因同源性高,蛋白结构相似,有望成为淋病奈瑟菌疫苗的候选抗原。
[Objective] To study the NspA gene of Neisseria Gonorrhoeae from standard strains （ggNG 29400 and NG 29403） and the characteristics of NspA fusion protein, so as to provide basis for the selection of target protein of gonorrhea vaccine. [Methods] The NspA gene fragments were obtained from two standard strains by PCR amplification and cloned into pET- 30c （gg＋） plasmid to generate NspA-pET-30c （gg＋） recombinants. The resulting recombinants were confirmed by Sac I and Hind III digestion and sequence analysis. Bioinformation analysis was conducted by using corresponding software and online sources. [ Results] NspA-pET-30c （gg＋） recombinants were constructed successfully and two expected bands of 0.5 kb and 5.4 kb were observed after digestion with Sac I and Hind Ⅲ. Sequences analysis confirmed the conservation of NspA among Neisseria Gonorrhoeae with the identity of 98% between two standard strains and available NspA gene and much similar to Neisseria meningitides. More than 40% of loop structure was observed in the predicted protein which might be the main function motifs. Similar tertiary structure was noticed in both NspA of Neisseria Gonorrhoeae and Neisseria meningitides. [Conclusion] The NspA gene of Neisseria Gonorrhoeae is of high conservation and much similar to Neisseria meningitides in both gene and protein, providing the possibility of NspA as a potential antigen for the vaccine of gonorrhea.
出处
《现代预防医学》
CAS
北大核心
2008年第22期4456-4460,4463,共6页
Modern Preventive Medicine
基金
四川大学科技创新基金项目(g2004CF02)
