摘要
【目的】研究重组鼠疫耶尔森氏菌(简称鼠疫菌)YscF蛋白的免疫原性。【方法】将含重组质粒YscF的E.coliBL21-YscF阳性克隆接种于LB液体培养基进行诱导表达;采用Ni2+亲和层析法对表达产物进行纯化;以蛋白体为佐剂吸附重组YscF蛋白,对Balb/c小鼠滴鼻免疫后,采用间接ELISA法检测血清中抗YscF的IgG和鼻、肺、小肠冲洗液中sIgA的抗体效价。【结果】重组YscF蛋白获得了高效表达,并以可溶性蛋白形式存在于上清中;滴鼻免疫小鼠后不但可以提高血清中总IgG效价,还可以诱导呼吸道分泌IgA型抗体;对腹腔注射100LD50强毒鼠疫菌攻击有一定保护效力,存活率为60%。【结论】YscF是鼠疫菌的一个有免疫原性的蛋白,能诱导较强的系统免疫和黏膜免疫;重组鼠疫菌YscF抗原可以作为改进的F1+V亚单位疫苗的辅助成分。
[Objective] Research on immunogenicity of plague recombinant protein YscF was conducted. [Method] The recombinant YscF protein of Yersinia pestis was expressed in E. coli BL21 cells,then it was purified by Ni^2＋ affinity chromatography. An proteosome- adsorbed vaccine of this YscF protein was intranarally immunized into experimental mice. The specific IgG and IgA to plague in sera, nasal solution, lung solution of the mice were detected by indirect ELISA. [Result] The recombined YscF protein could be expressed in the form of soluble protein at a high level. Intranasal immunization not only could increase the serum total IgG, but also induce secretory IgA antibodies of respiratory tract. It was able to induce an effective immunoprotection against intraperitoneal challenge of 100 LD50 ,and the survival rate was 60% . [Conclusion] Recombinant YscF has immunogenicity,can induce stronger systematic immune response and mucosal immune response,and is a possible alternative or adjunct protective vaccine of F1＋V against plague.
出处
《西北农林科技大学学报（自然科学版）》
CSCD
北大核心
2009年第12期68-72,共5页
Journal of Northwest A&F University(Natural Science Edition)
