淋病奈瑟菌候选外膜蛋白的免疫原性及膜定位的初步研究

A preliminary study on immunogenicity and membrane positioning of Neisseria gonorrhoeae candidate outer membrane proteins

目的对淋病奈瑟菌FA1090全基因组外膜蛋白进行免疫原性和膜定位的初步分析,以评价其作为候选疫苗的潜力。方法采用基因合成的方法构建重组质粒,转化感受态细胞,利用大肠杆菌表达系统对重组蛋白进行诱导表达,通过镍柱层析法纯化重组蛋白;用重组蛋白免疫BALB/c小鼠,制备免疫血清,通过间接ELISA检测抗体效价初步评价蛋白的免疫原性及其在外膜的定位。结果构建了15个重组质粒,获得了相应的纯化蛋白,通过免疫小鼠获得了免疫血清。免疫原性分析蛋白WP_003689500.1诱导小鼠产生相应IgG抗体的能力较强,膜定位分析显示有4个蛋白在外膜表达的可能性较高。结论蛋白WP_003689500.1诱导产生的抗体效价较高,与全细胞抗原的结合能力较强,可作为淋病奈瑟菌候选疫苗。

Objective A preliminary analysis of immunogenicity and membrane positioning of Neisseria gonorrhoeae FA1090 outer membrane proteins was carried out to evaluate their potential as candidate vaccines.Methods Using gene synthesis method to construct recombinant plasmids,transform receptor cells.The recombinant protein was induced by Escherichia coli expression system,and the recombinant protein was purified by nickel column tomography.The recombinant protein was obtained from the immune serum in BALB/c mice,and the antibody titer was detected by indirect ELISA to preliminarily evaluate the immunogenicity of the protein and its localization in the outer membrane.Results Fifteen recombinant plasmids were constructed,the corresponding purified proteins were obtained,and immune serums were obtained through immune mice.Immunogenic analysis found that protein WP_003689500.1 has a strong ability to induce mice to produce corresponding IgG antibodies,membrane positioning analysis found that 4 proteins in the outer membrane expression is more likely.Conclusion The antibody produced by protein WP_003689500.1 induction had high titer and strong binding ability with whole-cell antigens.It can be used as a candidate vaccine for N.gonorrhoeae.