表达HCV-NS3抗原的重组腺病毒载体疫苗的实验研究
发布时间：2018-10-11 07:42
【摘要】：目的 由于丙型肝炎病毒(hepatitis C virus,HCV)有多种基因型以及基因序列的高度变异性,使按传统方式研制HCV疫苗面临着诸多困难,而将编码目的抗原的外源基因导入受接种者的各类新型疫苗可望成为一条新的思路。以重组复制缺陷型腺病毒(replication-deficient recombinant adenovirus,RAd)为载体的疫苗,能有效地将外源基因导入宿主细胞内并使之高效表达,诱导机体产生针对目的抗原的免疫应答,因此,我们构建了可表达HCV非结构蛋白3(non-structural protein 3,NS3)的重组腺病毒RAd-NS3,并观察其免疫小鼠后,诱导机体产生特异性免疫应答的能力,为研制丙型肝炎疫苗探索新的途径。 方法 以真核表达质粒pRc/NS3为模板,应用聚合酶链反应(polymerase chain reaction,PCR)技术扩增编码NS3蛋白(302～935aa)的基因片段,定向克隆到重组腺病毒AdEasy-1系统的穿梭质粒
[Abstract]:Objective because hepatitis C virus (hepatitis C virus,HCV) has a variety of genotypes and high gene sequence variability, it is difficult to develop HCV vaccine in traditional way. The introduction of foreign gene encoding target antigen into inoculated new vaccines is expected to be a new way of thinking. The recombinant replication-deficient adenovirus (replication-deficient recombinant adenovirus,RAd) vaccine can effectively transfer foreign genes into host cells and express them efficiently, and induce the body to produce an immune response against the target antigen. We constructed the recombinant adenovirus RAd-NS3, expressing HCV nonstructural protein 3 (non-structural protein 3 NS3) and observed its ability to induce specific immune response after immunizing mice, and to explore a new approach for the development of hepatitis C vaccine. Methods using eukaryotic expression plasmid pRc/NS3 as template, the gene fragment encoding NS3 protein (302~935aa) was amplified by polymerase chain reaction (polymerase chain reaction,PCR) and cloned into shuttle plasmid of recombinant adenovirus AdEasy-1 system.
【学位授予单位】：四川大学
【学位级别】：博士
【学位授予年份】：2005
【分类号】：R392
本文编号：2263396
[Abstract]:Objective because hepatitis C virus (hepatitis C virus,HCV) has a variety of genotypes and high gene sequence variability, it is difficult to develop HCV vaccine in traditional way. The introduction of foreign gene encoding target antigen into inoculated new vaccines is expected to be a new way of thinking. The recombinant replication-deficient adenovirus (replication-deficient recombinant adenovirus,RAd) vaccine can effectively transfer foreign genes into host cells and express them efficiently, and induce the body to produce an immune response against the target antigen. We constructed the recombinant adenovirus RAd-NS3, expressing HCV nonstructural protein 3 (non-structural protein 3 NS3) and observed its ability to induce specific immune response after immunizing mice, and to explore a new approach for the development of hepatitis C vaccine. Methods using eukaryotic expression plasmid pRc/NS3 as template, the gene fragment encoding NS3 protein (302~935aa) was amplified by polymerase chain reaction (polymerase chain reaction,PCR) and cloned into shuttle plasmid of recombinant adenovirus AdEasy-1 system.
【学位授予单位】：四川大学
【学位级别】：博士
【学位授予年份】：2005
【分类号】：R392
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相关期刊论文 前2条
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