| 汉坦病毒H8205部分核壳蛋白基因在E.coli中的表达 |
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| 引用本文： | 沈昌贤,李钟铎,杨瑞馥,祝庆余.汉坦病毒H8205部分核壳蛋白基因在E.coli中的表达[J].中国病毒学,1999,14(2):geMap1-PH04. |
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| 作者姓名： | 沈昌贤 李钟铎 杨瑞馥 祝庆余 |
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| 作者单位： | 军事医学科学院微生物流行病研究所,北京100850 |
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| 摘 要： | 根据汉坦病毒H8205株NP基因的序列,设计一对引物,扩增NP前292个氨基酸多肽基因片段,克隆于表达载体pGEX-3X,与载体中26kD的谷胱苷肽巯基转移酶(GST)融合表达.SDS-PAGE显示表达产物(GST-NP)主要以包涵体形式存在.Western-blotting表明此融合蛋白有抗原性.包涵体经分离、洗涤、溶解后,Sepharose 6B层析纯化,用此融合蛋白作抗原,进行ELISA法检测临床HFRS病人标本的IgG和IgM,有很好的特异性和敏感性.有生物活性的汉坦病毒H8205 NP的体外表达成功,为汉坦病毒基因工程抗原的大量制备奠定了基础,也为汉坦病毒的临床检测和流行病学调查提供了一种廉价、安全、可靠的抗原.
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| 关 键 词： | 汉坦病毒，基因工程核衣壳蛋白，酶联免疫吸附试验(ELISA) |
| 修稿时间： | 1998－03－24 |
Expression of the S genome segment of Hantavirus H8205 and Application of the Expressed Polypeptide for Diagnosis of HFRS |
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| Shen Changxian,Li Zhongduo,Yang Ruifu,Zhu Qingyu.Expression of the S genome segment of Hantavirus H8205 and Application of the Expressed Polypeptide for Diagnosis of HFRS[J].Virologica Sinica,1999,14(2):geMap1-PH04. |
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| Authors: | Shen Changxian Li Zhongduo Yang Ruifu Zhu Qingyu |
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| Abstract: | The S genome segment encoding the
nucleocapsid protein (truncated) of Hantavirus H8205 was amplified by PCR and cloned into
the expression vector pGEX-3X and expressed in E coli. SDS-PAGE showed that the expressed
fusion protein (GST-NP) was mainly in the form of inclusion body. The expressed protein
was used as diagnostic antigen in solid-phase enzyme immunoassay. The assay was used to
detect IgG-and IgM-antibody in sera of HFRS patients originated from different geographic
regions of China. The results revealed highly sensitive and specific. The successful
expression and application of recombinant nucleocapsid protein of Hantavirus provides
cheap, safe and sensitive antigen for the diagnosis of HFRS in China. |
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