【摘要】 目的旨在获得无转化活性而保留抗原性的人乳头瘤病毒１６型（ＨＰＶ１６）Ｅ７Ｃ端，为进一步研制抗ＨＰＶ１６疫苗打下基础。方法应用自行设计合成的引物进行ＰＣＲ扩增ＨＰＶ１６Ｅ７Ｃ端亚基因片段，并克隆入真核表达载体ｐＬＮＣＸ。结果准确获得含有Ｅ７Ｃ亚基因片段的重组质粒ｐＬＮＣＥ７Ｃ，将ｐＬＮＣＥ７Ｃ转染Ｂ１６细胞。对Ｓｏｕｔｈｅｒｎ杂交阳性克隆应用Ｅ７多抗进行免疫组织化学方法检测，检出Ｅ７Ｃ的表达。结论所构建的表达质粒适于ＤＮＡ疫苗的制备。更多还原

【Abstract】 To develop vaccine against HPV16 is the hotspot in this field, this study was topave the rudiment for this purpose. Methods E7C subgene (encoding aa39 to aa98) was amplified by PCR, and then cloned into PLNCX plasmid. Results The recombinant PLNCE7C was obtained and identified by gel analysis of the restricted-endonuclease-digested fragments. It wasshown that PLNCE7C was expressed in transfected B16 cells. The expression product was identified in the positive cells by Southern blotting and immunohistochemistry method. It was localizedmainly in cytoplasm of B16 cells. Conclusions The plasmid PLNCE7C containing subgene ofHPV16E7C not only remains the antigenests of E7 gene but also eliminates the transforming activity of it. Meanwhile the expression of PLNCE7C is quite good. Therefore the plasmidpLNCE7C is suitable to develop DNA vaccine against HPV16.更多还原