大肠杆菌热稳定肠毒素Ⅰ和卵清蛋白融合蛋白的表达及免疫原性分析  

Expression and Immunogenicity Analysis of 3 × STa-ovalbumin Fusion Protein

作　　者：王录军[1] 段强德[1] 冯丽丽 WANG Lujun1,DUAN Qiangde1,FENG Lili2(1. Weinan Vocational and Technical College, Weinan 714026, China;2. Institute of Agricultural Economics and Information, Henan Academy of Agricultural Scienccs, Zhengzhou 450002, Chin)

机构地区：[1]渭南职业技术学院,陕西渭南714026 [2]河南省农业科学院农业经济与信息研究所,河南郑州450002

出　　处：《河南农业科学》2018年第4期141-144,共4页Journal of Henan Agricultural Sciences

基　　金：渭南市科技计划项目(2017JCYJ-4-4)

摘　　要：为探究大肠杆菌热稳定肠毒素Ⅰ(Heat-stable toxin typeⅠ,STa)和鸡卵清蛋白(Chicken ovalbumin)串联表达的3×STa-ovalbumin重组融合蛋白的免疫原性,原核表达融合蛋白后,经SDS-PAGE和Western blot方法鉴定,将表达的融合蛋白纯化后免疫小鼠,利用ELISA方法测定免疫小鼠血清中抗STa抗体的滴度,应用体外中和试验检测抗体对STa毒素的中和作用。SDSPAGE检测结果显示,融合蛋白分子质量约42 ku;Western blot分析结果显示,融合蛋白可被STa阳性血清特异性识别。小鼠免疫试验结果表明,融合蛋白具有较强的免疫原性,诱导机体产生的抗STa抗体滴度达到2.33,并且产生的抗体能在体外中和STa的毒性作用。以上结果表明,3×STaovalbumin融合蛋白具有良好的免疫原性。To evaluate the immunogenicity of 3 × STa-ovalbumin fusion protein,the 3 × STa-ovalbumin was expressed in E. coli BL21 and verified by SDS-PAGE and Western blot. The titers of anti-STa antibody ofserum from the mice immunized with 3 × STa-ovalbumin fusion protein were detected by ELISA,and the neutralization activity of anti-STa antibody was tested by STa neutralization assay. SDS-PAGE result showed that the fusion protein was proved to be approximately 42 ku of molecular mass. Western blot assayindicated that the fusion protein could be specifically recognized by anti-STa serum. Furthermore,anti-STa serum titers were detected at 2. 33 from the immunized mice,and the anti-STa antibodies possessed neutralization activity in vitro. The results indicated the expressed fusion protein had a good immunogenicity,which could be a candidate for ETEC subunit vaccine.

关 键 词：产肠毒素性大肠杆菌 STa毒素 融合蛋白 免疫原性 疫苗 

分 类 号：S855.1[农业科学—临床兽医学]