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单纯疱疹病毒 1 ICP27 蛋白:其表达、纯化和特异性抗血清的制备。

Herpes simplex virus type 1 ICP27 protein: its expression, purification and specific antiserum production.

机构信息

State Key Laboratory, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China.

出版信息

Virol Sin. 2010 Jun;25(3):199-205. doi: 10.1007/s12250-010-3116-2. Epub 2010 Jun 6.

Abstract

Herpes simplex virus type 1 (HSV-1) is the causative agent of cold sores and other more serious diseases. HSV-1 infected-cell protein 27 (ICP27) is an immediate-early regulatory phosphoprotein homologous to gene products identified in all classes of herpesviruses so far. To raise the antiserum to ICP27 for further characterization of its biological function, the ICP27 gene was cloned into the pET-28a (+) vector, then ICP27 protein was expressed in E. coli and purified by nickel-nitrilotriacetic acid (Ni(2+)-NTA) affinity resin column, finally the purified protein was used to raise antiserum. Western blot analysis demonstrated that the antiserum recognized the recombinant protein, and the antiserum was able to probe the ICP27 in HSV-1 infected cells with high specificity by immunofluorescence assay (IFA). Therefore, the specific antiserum will provide a valuable tool for further studies investigating ICP27's biological function during HSV-1 infection.

摘要

单纯疱疹病毒 1 型(HSV-1)是唇疱疹和其他更严重疾病的病原体。HSV-1 感染细胞蛋白 27(ICP27)是一种早期调节磷酸蛋白,与迄今为止所有疱疹病毒类别的基因产物同源。为了制备抗 ICP27 的血清,以进一步研究其生物学功能,将 ICP27 基因克隆到 pET-28a(+)载体中,然后在大肠杆菌中表达并通过镍-亚氨基二乙酸(Ni(2+)-NTA)亲和树脂柱纯化,最后用纯化的蛋白制备抗血清。Western blot 分析表明,该抗血清识别重组蛋白,并且该抗血清通过免疫荧光分析(IFA)能够特异性探测 HSV-1 感染细胞中的 ICP27。因此,该特异性抗血清将为进一步研究 ICP27 在 HSV-1 感染过程中的生物学功能提供有价值的工具。

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