单纯疱疹病毒2型感染细胞蛋白27重组真核表达质粒的构建及表达  

Construction and Expression of Eukaryotic Expression Plasmid Targeting Herpes Simplex Virus Type 2 Infection Cell Protein 27

作　　者：李慎秋[1] 梁宁[1] 赵红磊[2] 邹婕 谢春丽[1] 蒋思[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院皮肤科,武汉430030 [2]浙江大学医学院附属第一医院北仑分院皮肤科,宁波315806 [3]武汉市皮肤病防治研究所,武汉430031

出　　处：《华中科技大学学报（医学版）》2011年第6期719-723,共5页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：湖北省自然科学基金资助项目(No.2006ABA115)

摘　　要：目的构建单纯疱疹病毒2型(HSV-2)感染细胞蛋白27(ICP27)基因与pcDNA3.1(+)真核表达质粒的重组质粒pcDNA3.1(+)/ICP27。方法通过RT-PCR方法从HSV-2培养物中提取RNA制备ICP27的编码基因UL54DNA,将其与载体pcDNA3.1(+)连接。通过双酶切和测序鉴定重组质粒是否构建成功,Western blot法鉴定重组质粒在真核细胞COS-7中表达。结果重组质粒pcDNA3.1(+)/ICP27构建成功,可在COS-7中表达。结论成功构建了可在真核细胞内表达的pcDNA3.1(+)/ICP27重组质粒,为下一步研究该重组质粒作为DNA疫苗的免疫学效应奠定了基础。Objective To construct recombinant plasmid of herpes simplex virus type 2（HSV-2）virus-expressing infection cell protein 27（ICP27）gene and eukaryotic expression plasmid pcDNA3.1（＋）,pcDNA3.1（＋）/ICP27.Methods RNA was extracted from HSV-2 cultures by using RT-PCR to prepare UL54 DNA encoding ICP27,and the UL54 DNA was connected with the vector pcDNA3.1（＋）.The recombined plasmid was identified by restriction analysis and sequencing.The expression of recombinant plasmid was detected by using Western blot in eukaryotic cells COS-7.Results The recombinant plasmid pcDNA3.1（＋）/ICP27 was successfully constructed and could be expressed in COS-7 cells.Conclusion We successfully constructed a recombinant plasmid,pcDNA3.1（＋）/ ICP27,which can be expressed in eukaryotic cells,which lays a solid foundation for further studies on immunological effects of the recombinant plasmid as DNA vaccine.

关 键 词：单纯疱疹病毒2型 感染细胞蛋白27 重组质粒 DNA疫苗 

分 类 号：R349.6[医药卫生—基础医学]