人乳头瘤病毒16型E6和E7基因突变体表达蛋白的稳定性和抗原性分析  

Modification of HPV type 16 E6 and E7 genes,and analysis of stability and immunogenicity of the modified proteins

作　　者：职慧军[1] 韩立群[1] 任皎[1] 田厚文[1] 骆卫锋[1] 梁雨[1] 阮力[1] 

机构地区：[1]中国预防医学科学院病毒学研究所遗传免疫研究室,北京100052

出　　处：《中华实验和临床病毒学杂志》2002年第2期124-127,共4页Chinese Journal of Experimental and Clinical Virology

基　　金：国家"九五"科技攻关重点项目基金资助( 96 90 6 0 111)

摘　　要：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 select the mutants of HPV type 16 E6 and E7 genes suitable for construction of vaccine for treatment of cervical cancer.Methods E6 and E7 genes were modified by site\|directed mutagenesis.Several recombinant vaccina viruses were constructed by inserting the E6 or E7 mutants into the genome of vaccina virus Tiantan strain and employed to study their antigenicity.Results Western blot assay showed that the E6 mutant with substitution of Gly for Leu at amino acid site 50 and E7 mutant with substitution of Gly for Cys\|24 and Glu\|26 had no effect on their stability and antigenicity,but change of the Cys at position 91 of E7 dramatically reduced its stability and antigencity.Conclusion The results confirmed that the Zinic\|finger structure at the E7 C\|terminal plays an important role in the integrity and stability of E7 protein.

关 键 词：稳定性 抗原性分析 人乳头状瘤病毒 宫颈肿瘤 疫苗 插入诱变 

分 类 号：R373[医药卫生—病原生物学]