表達HCV-NS3抗原的重組腺病毒載體疫苗的實驗研究
發(fā)布時間:2018-10-11 07:42
【摘要】:目的 由于丙型肝炎病毒(hepatitis C virus,HCV)有多種基因型以及基因序列的高度變異性,使按傳統(tǒng)方式研制HCV疫苗面臨著諸多困難,而將編碼目的抗原的外源基因導入受接種者的各類新型疫苗可望成為一條新的思路。以重組復制缺陷型腺病毒(replication-deficient recombinant adenovirus,RAd)為載體的疫苗,能有效地將外源基因導入宿主細胞內并使之高效表達,誘導機體產生針對目的抗原的免疫應答,因此,我們構建了可表達HCV非結構蛋白3(non-structural protein 3,NS3)的重組腺病毒RAd-NS3,并觀察其免疫小鼠后,誘導機體產生特異性免疫應答的能力,為研制丙型肝炎疫苗探索新的途徑。 方法 以真核表達質粒pRc/NS3為模板,應用聚合酶鏈反應(polymerase chain reaction,PCR)技術擴增編碼NS3蛋白(302~935aa)的基因片段,定向克隆到重組腺病毒AdEasy-1系統(tǒng)的穿梭質粒
[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.
【學位授予單位】:四川大學
【學位級別】:博士
【學位授予年份】:2005
【分類號】:R392
本文編號: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.
【學位授予單位】:四川大學
【學位級別】:博士
【學位授予年份】:2005
【分類號】:R392
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相關期刊論文 前2條
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