HPV16 L1基因的原核表达及免疫反应性鉴定  

Prokaryotic expression of HPV 16 L1 gene and identification of its immune activity

作　　者：郝东风[1] 马正海[1] 王艳萍[1] 热西旦[1] 张富春[1] 

机构地区：[1]新疆大学生命科学与技术学院新疆生物资源基因工程重点实验室,新疆乌鲁木齐830046

出　　处：《细胞与分子免疫学杂志》2005年第4期428-431,共4页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金项目(No.39960079);新疆维吾尔自治区自然科学基金项目(No.200221103);教育部重点项目(No.02171)

摘　　要：目的:在大肠杆菌中表达人乳头瘤病毒16型(HPV16)主要衣壳蛋白L1,并鉴定其免疫反应性。方法:将HPV16L1基因克隆入原核表达载体pThioHisC中,构建重组表达载体。以重组载体分别转化大肠杆菌Top10和DH5α,在IPTG诱导下表达外源基因,用SDSPAGE和Westernblot对表达产物进行鉴定和分析。结果:构建了HPV16L1基因的原核表达质粒pThioHisC/HPV16L1,并在大肠杆菌中表达出相对分子质量(Mr)约为70800的蛋白。表达的蛋白能与抗HPV16L1抗体发生特异性反应。结论:在原核细胞中成功地表达HPV16L1基因,为HPV16L1疫苗的研制提供了必要的基础。AIM: To express the major capsid protein of human papillomavirus type 16 L1(HPV 16 L1) in E.coli and identify its immune activity. METHODS: The L1 gene of HPV 16 was cloned into the expression vector pThioHisC. The recombinant expression vector was transformed into E.coli , and the HisC-L1 protein was expressed under IPTG induction. The fusion protein was characterized by SDS-PAGE and Western blot. RESULTS: The HPV16-L1 gene in plasmid pThioHisC was expressed in E.coli as a fusion protein with M_r about 70 800. The fusion protein reacted specifically with antibodies against HPV16-L1. CONCLUSION: The HPV-16 L1 gene was expressed successfully in E.coli, which provides necessary basis for preparing HPV16 L1 vaccine in human.

关 键 词：人乳头状瘤病毒16型 主要衣壳蛋白 原核表达 免疫反应性 

分 类 号：R392.12[医药卫生—免疫学]