共表达HPV16L1、L2、E67蛋白的重组非复制型痘苗病毒的构建

CONSTRUCTION AND IDENTIFICATION OF NON-REPLICATION RECOMBINANT VACCINIA VIRUS CO-EXPRESSING HPV TYPE 16 L1/L2/E6/E7 PROTEINS

目的:构建共表达人乳头瘤病毒16型(HPV16)L1、L2、E67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、E67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、E67蛋白。结论:非复制型重组痘苗病毒NTVJE67CKL1L2可作为预防和治疗HPV16相关肿瘤及其癌前病变的候选疫苗。

Objective:To generate an HPV16 prophylactic and theraperutic vaccine candidate for cervical cancer.Methods:HPV16 major capsid protein L1 gene/minor capsid protein L2 gene and HPV16 early E6/E7 genes were inserted into a vaccinia virus expression vector. A strain of non-recombinant vaccinia virus containing the sequences was obtained through a homologous recombination and identification.Result:DNA hybridization confirmed that the HPV16L1/L2/E67 genes were integrated into vaccinia virus DNA . Western blot result showed that full-length L1/L2/E67 proteins were co-expressed in CEF cells infected with the recombinant virus.Conclusion:NTVJE67CKL1L2 could be taken as a candidate of prophylactic and theraperutic vaccine for HPV-associated tumors and their precancerous transformations.