HPV16 L1基因的原核表达及免疫反应性鉴定

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HPV-16L1基因克隆人原核表达载体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SDS—PAGE和Western blot对表达产物进行鉴定和分析。结果：构建了HPV—16L1基因的原核表达质粒pThioHisC／HPV—16L1，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HPV—16L1抗体发生特异性反应。结论：在原核细胞中成功地表达HPV—16L1基因，为HPV—16L1疫苗的研制提供了必要的基础。

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

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.