PVAX1-HPV58mE6E7FcGB复合基因疫苗的构建及真核表达  

Construction and Eukaryotic Expression of PVAX1-HPV58mE6E7FcGB Composite Gene Vaccine

作　　者：王鹤[1] 于继云[2] 李力[1] 

机构地区：[1]广西医科大学附属肿瘤医院,南宁530021 [2]军事医学科学院基础医学研究所,北京100850

出　　处：《生物医学工程学杂志》2013年第5期1102-1107,共6页Journal of Biomedical Engineering

基　　金：国家自然科学基金资助项目(81260386);广西卫生厅重点课题资助项目(重200970)

摘　　要：为构建和表达人乳头瘤病毒(HPV)58型相关宫颈癌治疗用疫苗,将HPV58mE6E7融合基因片段插入真核表达载体pCI-Fc-GPI中,构建重组真核表达质粒pCI-sig-HPV58mE6E7-Fc-GPI。再将得到的sigHPV58mE6E7-Fc-GPI片段插入新型疫苗载体PVAX1-IRES-GM/B7中,构建PVAX1-HPV58mE6E7FcGB复合基因疫苗。经限制性内切酶鉴定及测序分析证实PVAX1-HPV58mE6E7FcGB复合基因疫苗构建成功。流式细胞术及免疫荧光检测表明该疫苗中的融合抗原sig-HPV58mE6E7-Fc-GPI及分子佐剂GM-CSF/B7.1能够同时实现真核表达。表明PVAX1-HPV58mE6E7FcGB复合基因疫苗可作为治疗HPV58相关肿瘤及其癌前病变的候选疫苗。To construct and express a composite gene vaccine for human papillomavirus 58（HPV58）-associated cervi- cal cancer, we inserted HPV58mE6E7 fusion gene into pCI-Fc-GPI eukaryotic expression vector, constructing a re- combinant plasmid named pCI-sig-HPV58mE6E7-Fc-GPI. Then we further inserted fragment of sig-HPV58mE6E7- Fc GPI into the novel vaccine vector PVAXI-IRES-GM/BT, constructing PVAX1-HPV58mE6E7FcGB composite gene vaccine. PVAXl-HPV58mE6E7FcGB vaccine was successfully constructed and identified by restriction endonu- clease and sequencing analysis. Eukaryotic expression of fusion antigen sig HPV58mE6E7-Fc-GPI and molecular ad- juvant GM-CSF and B7.1 were proved to be realized at the same time by flow cytometry and immunofluorescence. So PVAX1-HPV58mE6E7FcGB can be taken as a candidate of therapeutic vaccine for HPV58-associated tumors and their precancerous transformations.

关 键 词：人乳头瘤病毒58型 E6基因 E7基因 DNA疫苗 真核表达 

分 类 号：R737.33[医药卫生—肿瘤]