His-tag不影响RSV重组蛋白G1F/M2的免疫原性  

His-tag does not change immunogenicity of recombinant protein G1F/M2 of respiratory syncytial virus

作　　者：曾瑞红[1] 王卫华[2] 房桂珍[2] 龚伟[3] 梅兴国[3] 魏林[1] 

机构地区：[1]河北医科大学免疫教研室,石家庄050017 [2]河北医科大学生物医学工程中心,石家庄050017 [3]军事医学科学院毒物药物研究所,北京100850

出　　处：《中国免疫学杂志》2008年第12期1066-1070,共5页Chinese Journal of Immunology

摘　　要：目的:观察His-tag是否影响RSV重组蛋白G1F/M2的免疫原性。方法:PCR扩增G1和F/M2基因片段,插入表达载体pET-His和pET-DsbA-His中,转化E.coli BL21(DE3),IPTG诱导表达,采用Ni+螯合亲和层析法纯化得His-G1F/M2和DsbA-His-G1F/M2,将后者用凝血酶消化,再经Ni+螯合亲和层析法纯化得G1F/M2,将His-G1F/M2和G1F/M2免疫BALB/c小鼠,用ELISA测定抗体滴度,MTT法测定细胞毒性T细胞活性(CTL)。结果:两种蛋白在BALB/c小鼠中诱导的RSV特异性抗体和CTL活性无显著差异。结论:His-tag不影响RSV重组蛋白G1F/M2的免疫原性。Objective：To investigate whether His-tag to change the immunogenicity of recombinant protein G1F/M2 of respiratory syncytial virus.Methods：The G1 and F/M2 gene fragments were amplified by PCR method and then ligated into the expressing vector pET-His or pET-DsbA-His.Each recombinant plasmid was transferred into E.coli BL21（DE3） and the expression was induced by IPTG.The expressed His-G1F/M2 or DsbA-His-G1F/M2 was purified by affinity chromatography.The latter was digested with thrombase and G1F/M2 was purified by affinity chromatography.His-G1F/M2 or G1F/M2 was used to immunize BALB/c mice.Anti-RSV antibody was measured by ELISA and RSV-specific CTL responses by MTT.Results：No significant difference was observed between the level of anti-RSV antibody or RSV-specific CTL response induced by G1F/M2 and that by His-G1F/M2.Conclusion：His-tag does not change the immunogenicity of recombinant protein G1F/M2 of respiratory syncytial virus.

关 键 词：呼吸道合胞病毒 HIS-TAG 重组蛋白G1F/M2 免疫原性 

分 类 号：R392.11[医药卫生—免疫学]