丙肝病毒复合多表位抗原与乙肝病毒核心抗原融合基因的克隆及表达

发布时间：2018-05-21 04:09

本文选题：丙肝病毒表位抗原 + 乙肝病毒核心抗原　；参考：《吉林大学》2007年硕士论文

【摘要】： 本研究为丙肝核酸疫苗的基础性研究工作。应用重叠延伸PCR技术,获得了HCV复合多表位抗原与HBV核心抗原(HBcAg)的融合基因,命名为CMCe。实验进行过程中,设立HBc基因(C基因)平行对照组。将CMCe定向克隆于pET-28a(+)原核表达载体,构建原核表达质粒pET-28a(+)-CMCe,转化入大肠杆菌DH5α感受态细胞中,测序正确后筛选阳性克隆,转化至BL21(DE3)宿主菌中,利用IPTG诱导目的基因在大肠杆菌中表达。经过western blot分析后,表达产物能与慢性丙型肝炎病人血清中的抗体发生特异性的抗原-抗体反应。表明融合基因所表达的蛋白有良好的免疫原性。
[Abstract]:This study is the basic research work of hepatitis C nucleic acid vaccine. The fusion gene of HCV complex polyepitope antigen and HBV core antigen HBcAgwas obtained by overlapping extension PCR technique and named CMCe. During the course of the experiment, HBc gene C gene was set up in parallel control group. CMCe was cloned into pET-28a () prokaryotic expression vector, and the expression plasmid pET-28a (pET-28a) was transformed into Escherichia coli DH5 伪 competent cells. IPTG was used to induce the expression of the target gene in Escherichia coli. After western blot analysis, the expressed product could react with antibody in serum of patients with chronic hepatitis C. The results showed that the protein expressed by the fusion gene had good immunogenicity.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R392;Q78

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