人类8型疱疹病毒K8.1N基因编码包膜糖蛋白的原核表达及其抗原特异性分析  

The prokaryotic expression of HHV-8 envelope glycoprotein K8.1N gene and analysis of its antigenic specification

作　　者：赵淑君[1] 何方平[2] 王星[3] 陈晓[1] 何斌[4] 温浩[3] 

机构地区：[1]新疆医科大学基础医学院病理解剖学教研室,新疆乌鲁木齐830054 [2]新疆医科大学第一附属医院肝病中心,新疆乌鲁木齐830054 [3]新疆医科大学第一附属医院中心实验室,新疆乌鲁木齐830054 [4]美国芝加哥大学伊利诺医学院免疫与微生物系病毒中心实验室,美国芝加哥il606127344

出　　处：《新疆医科大学学报》2007年第2期97-100,共4页Journal of Xinjiang Medical University

基　　金：新疆维吾尔自治区自然科学基金资助项目(200421122)

摘　　要：目的:制备人类8型疱疹病毒(HHV-8)K8.1N基因编码包膜糖蛋白的原核表达融合蛋白,并鉴定其抗原特异性。方法:将重组质粒pET41a-K8.1N转化大肠杆菌BL21感受态细胞,异丙基硫代-β-D-半乳糖苷(IPTG)诱导表达谷胱甘肽S转移酶(GST)/K8.1N融合蛋白,经Glutathione Sepharose4B亲和层析柱纯化。采用Western blot法观察GST/K8.1N融合蛋白(作为抗原)与新疆地区、法国卡波西肉瘤(KS)患者血清以及四川健康儿童血清的血清学反应。结果:IPTG诱导后的菌体裂解蛋白,经12%聚丙烯酰胺凝胶电泳(SDS-PAGE)后,显示有一个46kD的融合蛋白表达,经Westernblot法鉴定此融合蛋白能被KS患者血清特异性地识别,新疆地区KS患者血清检出率为78.6%,法国KS为12.0%,四川健康儿童血清无一例与GST/K8.1N融合蛋白发生反应。结论:HHV-8包膜糖蛋白编码基因在大肠杆菌BL21中获得正确表达,并与KS患者血清具有特异识别性。Objective： To express the envelope glycoprotein K8. 1N coding gene of human herpesvirus 8 （HHV-8） in E. coli , and further identify its antigenic specification. Methods. The recombinant plasmid pET41a-KS. 1N was transferred to E. coli competent cells BL21 （DH3） and the expression of the GST/KS. 1N fusion protein was induced by IPTG. The fusion protein was further purified with glutathione sepharose 4B affinity chromatography. Its serologic reaction with the sera of Xinjiang Kaposi＇s sarcoma （KS）, French KS patients and Sichuan healthy children was observed by Western blot. Results： A 46 KD of fusion protein was exhibited on a 12% of sodium dodecyl sulfate-polyacrylamide gel electrophoresis （SDS-PAGE） when the recombinant bacteria was induced by IPTG. The fusion protein was recognized by KS patient sera in Western blot, the detection rate of Xinjiang was 78.6%, French was 12.0% and the specific reaction wasn＇t observed in Sichuan healthy children. Conclusion： The envelope glycoprotein KS. 1N coding gene of HHV-8 was correctly expressed in E. coli BL21 cells. Western blot analysis demonstrated that the fusion protein was specifically recognized by KS patient sera.

关 键 词：人类疱疹病毒8型 包膜糖蛋白K8.1N基因 原核表达 抗原特异性分析 

分 类 号：R733.3[医药卫生—肿瘤]