乙型肝炎病毒多表位抗原基因真核表达载体的构建及其在哺乳动物细胞中的表达

将自行设计、合成的乙型肝炎病毒多表位抗原基因BPT ,克隆入真核表达载体pCDNA3 1,构建重组质粒pCDNA3 1/BPT。后者经脂质体法转染小鼠BALB/c 3T3细胞 ,G4 18加压筛选出阳性转染细胞 ,并用间接免疫荧光法鉴定其在小鼠BALB/c 3T3细胞的表达。该重组质粒经胫骨前肌注射小鼠 ,10 0 μg/次每只小鼠 ,间隔 2周加强一次 ,共 3次。免疫诱发了特异性抗体和淋巴细胞增殖反应 ,并用PCR法在小鼠的心脏、肝脏及肺组织中检测到pCDNA3 1/BPT ,这为研究HBV多表达抗原的核酸疫苗提供了初步的资料

Construction of Eukaryotic Expression Vector for Multi-Epitope Gene of Hepatitis B Virus and Its Expression in Mammalian Cells

The self-designed and synthesized mul ti-epitope gene BPT of hepatitis B virus(HBV) was cloned into eukaryotic expression vector pcDNA3.1 to construct the recombinant plasmid pcDNA3.1-BPT. The recombinant plasmid was transfected into BALB/c 3T3 cells by liposome method, and the positive transfected cells were screened through G418. By using the indirect immunofluorescence technique, its expression in BALB/c 3T3 cells was identified. This recombinant plasmid was injected into mice through anterior tibia muscle, 100 μg per mouse, and mice were challenged three times with two weeks interval. It was found that the immunization with this recombinant plasmid could induce the specific antibody responses and the proliferative responses of lymphocytes. The recombinant plasmid pcDNA3.1-BPT could be detected in heart, liver and lung tissues of mice by means of PCR. These results provide a preliminary data for the further study of multi-epitope gene nucleic acid vaccine.