| His-tag不影响RSV重组蛋白G1F/M2的免疫原性 |
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| 引用本文： | 曾瑞红,王卫华,房桂珍,龚伟,梅兴国,魏林.His-tag不影响RSV重组蛋白G1F/M2的免疫原性[J].中国免疫学杂志,2008,24(12). |
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| 作者姓名： | 曾瑞红 王卫华 房桂珍 龚伟 梅兴国 魏林 |
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| 作者单位： | [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 |
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| 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。
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| 关 键 词： | 呼吸道合胞病毒 His-tag 重组蛋白G1F/M2 免疫原性 |
His-tag does not change immunogenicity of recombinant protein G1F/M2 of respiratory syncytial virus |
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| ZENG Rui-Hong,WANG Wei-Hua,FANG Gui-Zhen,GONG Wei,MEI Xing-Guo,WEI Lin.His-tag does not change immunogenicity of recombinant protein G1F/M2 of respiratory syncytial virus[J].Chinese Journal of Immunology,2008,24(12). |
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| Authors: | ZENG Rui-Hong WANG Wei-Hua FANG Gui-Zhen GONG Wei MEI Xing-Guo WEI Lin |
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| Affiliation: | ZENG Rui-Hong,WANG Wei-Hua,FANG Gui-Zhen,GONG Wei,MEI Xing-Guo,WEI Lin.Department of Immunology,Hebei Medical University,Shijiazhuang 050017,China |
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| Abstract: | 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 purif... |
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| Keywords: | Respiratory syncytial virus His-tag Recombinant protein G1F/M2 Immunogenicity |
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