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.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
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)
