表达HPV16 L1、L2E7蛋白的非复制重组痘苗病毒的构建和鉴定

CONSTRUCTION AND IDENTIFICATION OF NON-REPLICATING RECOMBINANT VACCINIA VIRUS EXPRESSING HPV16 L1,L2E7 PROTEINS

目的:构建防治宫颈癌的表达人乳头瘤病毒16型、HPV16)L1、L2E7蛋白的非复制重组痘苗病毒疫苗株。方法:以痘苗病毒为载体,利用同源重组技术筛选共表达HPV16L1、L2E7蛋白的重组痘苗病毒,用PCR和Western-blot技术进行鉴定。结果:该病毒在鸡胚成纤维细胞(CEF)中连续传代15代,经PCR检测证实病毒基因组中有HPV16L1、L2E7目的基因插入,Western-blot检测证实病毒可以稳定表达L1、L2E7蛋白。结论:构建的病毒NTVJL1L2E7可以在真核细胞中表达HPV16L1、L2E7蛋白,可以作为治疗和预防HPV16相关疾病的候选疫苗。

Objective：To construct a non-replicating vaccinia virus expressing HPV16 L1, L2E7 proteins as a candidate vaccine for cervical cancer. Methods： Using vaccinia virus vector, we generated a strain of nonreplicating recombinant vaccinia virus vaccine expressing HPV16 L1, L2E7 proteins by homologous recombination and identified by PCR and Western-hloting. Result：We demonstrated that the L1, L2E7 gene of HPV16 were integrated into vaccinia genosome and could express L1, L2E7 protein stably when infecting the CEF using PCR and Western-blot assay. Conclusion： NTVJLI/L2E7 can express L1, L2E7 protein of HPV16 and can be taken as a candidate vaccine for HPV16 associated diseases.