甲肝病毒表位抗原与乙肝病毒核心抗原融合基因表达的研究

发布时间：2018-03-19 21:27

本文选题：甲肝病毒表位抗原　切入点：乙肝病毒核心抗原　出处：《吉林大学》2005年硕士论文　论文类型：学位论文

【摘要】：本研究为甲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HAV 表位抗原与HBV 核心抗原(HBcAg)的融合基因,命名为CVPX。实验进行过程中,设立HBc 基因(C 基因)平行对照组。将CVPX定向克隆于pET-28a(+) 原核表达载体, 构建原核表达质粒pET-28a(+)-CVPX,转化入大肠杆菌DH5α感受态细胞中,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,表达产物以包涵体形式存在。表达产物能与抗-HAV 抗体发生特异性的抗原-抗体反应。同时将CVPX定向克隆于pVAX1 真核表达载体,成功构建真核表达质粒pVAX1-CVPX,利用脂质体转染CHO 细胞,mRNA 水平上检测到目的基因进行了有效转录,蛋白质水平上检测到表达的蛋白具有HAV 抗原的反应原性。
[Abstract]:In this study, the fusion gene of HAV epitope antigen and HBV core antigen HBcAgwas obtained by using overlapping extended PCR technique. The fusion gene was named CVPX during the experiment. The CVPX was cloned into the prokaryotic expression vector pET-28a (pET-28a) and transformed into Escherichia coli DH5 伪 competent cells. IPTG was used to induce the expression of the target gene in Escherichia coli, and the expression product was in the form of inclusion body. The expressed product could react with the anti-HAV antibody specifically. At the same time, the CVPX was cloned into the eukaryotic expression vector of pVAX1. The eukaryotic expression plasmid pVAX1-CVPXwas successfully constructed, and the target gene was detected by liposome transfection into CHO cells for effective transcription. At the protein level, the expressed protein showed the reactivity of HAV antigen.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R392.1

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