共表达HPV16L1、L2、E6、E7蛋白的重组非复制型痘苗病毒构建及鉴定  

Construction and identification of non-replication recombinant vaccinia virus co-expressing human papillomavirus type 16 L1/L2/E6/E7 proteins

作　　者：黄薇[1] 田厚文[2] 任皎[2] 范江涛[2] 赵莉[2] 边涛[2] 陆振华 阮力[2] 

机构地区：[1]广西医科大学肿瘤医院妇瘤科,南宁530021 [2]中国疾病预防控制中心病毒病预防控制所应急技术中心

出　　处：《中华实验和临床病毒学杂志》2005年第3期240-243,共4页Chinese Journal of Experimental and Clinical Virology

基　　金：国家高技术研究发展计划863项目资助(2002AA216041)

摘　　要：目的构建共表达人乳头瘤病毒16型(HPV16)L1、L2、E6、E7蛋白的非复制型重组痘苗病毒人用疫苗株。方法以痘苗病毒为载体、利用同源重组技术筛选共表达HPV16L1、L2、E6、E7蛋白的重组痘苗病毒并对其进行鉴定。结果该病毒在CEF细胞上连续传至第15代,经斑点杂交结果表明重组病毒基因组中有L1、L2、E6、E7基因插入;经WesternBlot检测,重组病毒能稳定表达HPV16L1、L2、E6、E7蛋白。结论非复制型重组痘苗病毒NTVJE6E7CKL1L2可作为预防和治疗HPV16相关肿瘤及其癌前病变候选疫苗。Objective To generate a human papilomavirus （HPV16） prophylactic and therapeutic vaccine candidate for cervical cancer. Methods HPV16 major capsid protein L1 gene/minor capsid protein 12 gene and HPV16 early E6/E7 genes were inserted into a vaeeinia virus expression vector. A strain of non-recombinant vaeeinia virus containing the sequences was obtained through a homologous recombination and identified. Results DNA hybridization confirmed that the HPV16L1/L2/E6/E7 genes were integrated into vaeeinia virus DNA. Western Blot result showed that full-length L1/L2/E6/E7 proteins were coexpressed in CEF cells infected with the recombinant virus. Conclusion NTVJE6E7CKL1L2 could be taken as a candidate of prophylactic and therapeutic vaccine for HPV-associated tumors and their precancerous transformations.

关 键 词：乳头状瘤病毒 人 宫颈肿瘤 痘苗病毒 疫苗 非复制型重组痘苗病毒 非复制型痘苗病毒 HPV16L1 E7蛋白 共表达 E6 

分 类 号：R392[医药卫生—免疫学]