Expression of the S genom e segm ent of H antavirus H8205 and Application of the Expressed Polypeptide for Diagnosis of HFRS

CHEN Chang-Xian; LI Zhong-Duo; YANG Rui-Bi; CHU Qiang-Tu

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April 1999

CHEN Chang-Xian, LI Zhong-Duo, YANG Rui-Bi and CHU Qiang-Tu. Expression of the S genom e segm ent of H antavirus H8205 and Application of the Expressed Polypeptide for Diagnosis of HFRS[J]. Virologica Sinica, 1999, 14(2): 110-115.

Citation: CHEN Chang-Xian, LI Zhong-Duo, YANG Rui-Bi, CHU Qiang-Tu. Expression of the S genom e segm ent of H antavirus H8205 and Application of the Expressed Polypeptide for Diagnosis of HFRS .VIROLOGICA SINICA, 1999, 14(2) : 110-115.

汉坦病毒H8205部分核壳蛋白基因在E.coli中的表达

1.
军事医学科学院散生物流行病研究所，北京100850

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ｂｌｏｔｉｎｇ表明此融合蛋白有抗原性。包涵体经分离、洗涤、溶解后，Ｓｅｐｈａｒｏｓｅ６Ｂ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Ｈ８２０５ＮＰ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。
- 汉坦病毒
- , 基因工程核衣壳蛋白
- , 酶联免疫吸附试验（ELISA）

Expression of the S genom e segm ent of H antavirus H8205 and Application of the Expressed Polypeptide for Diagnosis of HFRS

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.
- Han tavirus
- , Recombinant nucleocapsid protein
- , Enz yme-linked imm unosorhent assay(ELISA)

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Expression of the S genom e segm ent of H antavirus H8205 and Application of the Expressed Polypeptide for Diagnosis of HFRS

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.