人乳头瘤病毒16型E2蛋白真核载体的构建与表达

目的构建人乳头瘤病毒16型（HPV16）E2蛋白真核载体,为研究其基因疫苗免疫活性奠定实验基础。方法 PCR扩增HPV16 E2基因片段,将其连接到真核表达载体pcDNA3.1（＋）,双酶切及测序鉴定。将质粒pcDNA3.1（＋）与pcDNA3.1（＋）/HPV16 E2分别转染HeLa细胞,RT-PCR鉴定E2基因在HeLa细胞中的表达。结果 HPV16 E2基因片段插入到pcDNA3.1（＋）相同酶切位点,即构建了真核表达载体pcDNA3.1（＋）/HPV16 E2;转染pcDNA3.1（＋）/HPV16 E2的细胞,检测到HPV16 E2 mRNA。结论构建的真核表达载体pcDNA3.1（＋）/HPV16 E2能在HeLa细胞内有效表达。

Expression and Construction of Eukaryotic Vector of HPV16 E2 Protein

ObjectiveTo provide the experimental basis for further researching immunological competence of human papillomavirus type 16 (HPV16) E2 gene vaccine,eukaryotic expression vector of HPV16 E2 protein is constructed.MethodsHPV16 E2 gene fragment amplified by PCR was inserted into eukaryotic expression vector of pcDNA3.1(+) with the same restriction sites.Enzymes digestion and DNA sequencing were used to identify E2 gene fragment.The eukaryotic expression vector of pcDNA3.1(+) or pcDNA3.1(+)/HPV16 E2 was transfected into HeLa cells,respectively.The expression of HPV16 E2 was analyzed using RT-PCR.ResultsThe gene fragment of HPV16 E2 was successfully insert into pcDNA3.1(+).The mRNA products of HPV16 E2 were detected in the cells transfected with pcDNA3.1(+)/HPV16 E2.ConclusionEukaryotic expression vector of pcDNA3.1(+)/HPV16 E2 successfully express HPV16 E2 mRNA in HeLa cells.