中文摘要：

目的 制备人疱疹病毒7型（HHV-7）pp85蛋白及其抗体。方法 采用聚合酶链反应（PCR）方法自HHV-7型YY5分离株中扩增出pp85基因，经测序后构建原核表达质粒PGEX-6p-1＋ppSSb。重组质粒转化大肠埃希菌Rosetta，诱导蛋白表达。应用酶切鉴定、十二烷基硫酸钠．聚丙烯酰胺凝胶电泳（SDS-PAGE）及蛋白质印迹法等方法确保基因片段的正确性及表达蛋白的特异性。表达的蛋白经亲和层析纯化后，免疫动物制备多克隆抗体，并通过免疫荧光法鉴定抗体的特异性。结果 成功地获得了高纯度的pp85融合蛋白，纯度可达90％以上；免疫动物后制备的多克隆抗体效价可达1：102400，该抗体能特异识别HHV-7抗原，而不与HHV-6反应。结论 获得高纯度的HHV-7pp85融合蛋白及其高效价的抗体，将进一步应用于临床检验。

英文摘要：

Objective The goal of the current study was to express the pp85 gene of human herpesvirus 7 （HHV-7） and to prepare anti-pp85 polyclonal antibodies. Methods The pp85 gene of HHV-7 was amplified using PCR and this amplified gene was cloned into a prokaryotic expression vector and named as PGEX-6P-1 ＋ pp85b. The recombinant plasmid was then transformed into Escherichia coli Rosena. The accuracy of the inserted gene and the specificity of the proteins were verified by restriction enzyme digestion, SDS-PAGE, and Western blot. The expressed pp85 fusion protein was then purified using affinity chromatography and was used to immunize rabbits in order to obtain the anti- pp85 antibodies. The specificity of these antibodies was verified using immunofluorescence. Results The purity of the recombinant pp85 was above 90% and the titer of anti-pp85 antibodies was 1 ： 102 400. As an indication of the specificity of the antibodies produced, this polyclonal antibody was shown to react with HHV-7 infected cells but failed to react with human herpesviras 6 （HHV-6） infected cells as shown by IFA. Conclusion In the current study we were able to obtain a highly purified form of the HHV-7 pp85 fusion protein and the corresponding hightiter polyclonal antibodies were produced in rabbit.