人乳头瘤病毒16型E_6基因表达产物单克隆抗体的制备研究  

Expression of HPV_(16) E_6 Gene and Preparation of Monoclonal Antibody against the Expression Product

作　　者：桂长云[1] 宋国兴[1] 李昆[1] 司静懿[1] 韩日才[1] 贾丽萍[1] 马钧 

机构地区：[1]中国医学科学院,中国协和医科大学基础医学研究所,北京医科大学第一附属医院

出　　处：《中国医学科学院学报》1994年第4期290-295,共6页Acta Academiae Medicinae Sinicae

摘　　要：将含有人乳头瘤病毒１６型（ＨＰＶ_（１６））Ｅ_６基因的重组质粒ＰＡＳ１－ＨＰＶ_（１６）Ｅ_６转染大肠杆菌ＡＲ１２０，在萘啶酮酸诱导下进行表达，表达产物经分离、纯化和鉴定获得一种分子量为１９０００的Ｅ_６表达蛋白。用纯化后的Ｅ６蛋白免疫ＢＡＬＢ／ｃ小鼠，免疫脾细胞与骨髓瘤细胞ＳＰ２／０－Ａｇ１４在ＰＥＧ_（４０００）作用下进行融合，经ＨＡＴ培养基筛选杂交瘤细胞株，甲基纤维素半固体培养基克隆，克隆筛选，ＥＬＩＳＡ检测和再克隆，得到一株持续稳定分泌抗ＨＰＶ_（１６）Ｅ_６蛋白特异性单克隆抗体的杂交瘤株ＲＡＣ_６。The recombinant plasmid PAS1-HPV16 E6 containing the HPV16 E6 gene was expressed in E.coli AR120 under nalidixic acid induction.A 19KD expression protein was isolated,puri fied and identified. Mice were immunized with the purified E6 expression protein.A murine hybridoma, RAC6,was obtained by fusing splenic cells from an immunized BABL/c mouse with mouse myeloma cell line SP2/0-Ag14 cells, fellovved by screening in HAT medium,cloning and recloning in methyl cellulose.The hybridoma RAC6 stably produces specific monoclonal antibody against the E6 expression protein.

关 键 词：乳头瘤病毒 基因表达 子宫肿瘤 

分 类 号：R737.33[医药卫生—肿瘤]