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目的 筛选适合于宫颈癌治疗性疫苗研制的人乳头瘤病毒 1 6型 (HPV1 6)E6和E7突变基因。方法 采用定点诱变技术对E6和E7基因进行改造 ,得到几种E6和E7基因的突变体 ;构建成表达野生型和突变型E6与E7的重组痘苗病毒 ,对这些重组痘苗病毒表达的E6和E7蛋白的稳定性及其免疫原性进行了比较研究。结果 E6蛋白第 50位氨基酸的突变、E7蛋白第 2 4和 2 6位氨基酸的突变都不影响蛋白的稳定性和抗原性 ,但E7蛋白第 91位氨基酸的突变使E7蛋白的稳定性和抗原性大大减弱。结论 证实了E7蛋白C 末端的锌指结构对E7蛋白结构的完整性和稳定性是非常重要的。
Objective To screen human papillomavirus type 1 (HPV16) E6 and E7 mutant genes which are suitable for the therapeutic vaccine of cervical cancer. Methods The E6 and E7 genes were modified by site-directed mutagenesis, and several E6 and E7 gene mutants were obtained. Recombinant vaccinia virus expressing wild type and mutant E6 and E7 was constructed. The recombinant vaccinia virus E6 and E7 E7 protein stability and immunogenicity were compared. Results The amino acid mutation at position 50 of E6 protein and amino acid mutations at positions 2-4 and 26 of E7 protein did not affect the stability and antigenicity of the protein. However, mutation of amino acid 91 of E7 protein caused the stability of E7 protein and antigen Sex is greatly reduced. The conclusion confirms that the C-terminal zinc finger structure of E7 protein is very important for the structural integrity and stability of the E7 protein.