甲型流感病毒NA基因核酸疫苗的构建及表达

发布时间：2018-11-08 06:47

【摘要】： 流行性感冒是由流感病毒引起的人和多种动物的一种急性、热性、高度接触性呼吸道传染病,其流行范围广泛,可引起人和多种动物的发病,甚至死亡。传统的流感病毒灭活疫苗刺激机体产生IgG抗体,通过抗体介导的免疫来抵抗流感病毒的感染,流感病毒的漂移和转变常常使接种流感灭活疫苗失败,导致流感的流行。核酸疫苗以天然构象形式免疫,可刺激机体产生细胞和体液免疫双重免疫效果而达到抗病毒的作用,成为目前流感疫苗的研究热点。本研究将甲型流感病毒(H1N1)接种鸡胚后收获尿囊液,提取流感病毒总RNA,RT-PCR法扩增HA基因。经亚克隆后获得质粒pMD-T/NA,将其与质粒pVAX1分别双酶切后连接,构建出甲型流感病毒表达载体pVAX/NA,酶切鉴定表明成功构建出甲型流感病毒表达载体pVAX/NA,Western blot检测证明了流感病毒NA蛋白的表达。
[Abstract]:Influenza is an acute, feverish and highly contact respiratory infectious disease caused by influenza virus in humans and many animals. It has a wide epidemic range and can cause morbidity and even death of people and many animals. The traditional inactivated influenza virus vaccine stimulates the body to produce IgG antibody and resists the influenza virus infection through the antibody mediated immunity. The drift and transformation of the influenza virus often make the influenza inactivated vaccine fail and lead to the influenza epidemic. Nucleic acid vaccine immunized in the form of natural conformation can stimulate both cellular and humoral immunity to achieve the antiviral effect and become the research hotspot of influenza vaccine at present. In this study, the allantoic fluid of influenza A virus (H1N1) was harvested after inoculation of chicken embryo, and the HA gene was amplified by total RNA,RT-PCR of influenza virus. After subcloning, the plasmid pMD-T/NA, was digested with the plasmid pVAX1, respectively, and ligated with the plasmid pVAX1. The expression vector of influenza A virus, pVAX/NA, was constructed by restriction endonuclease digestion. The results showed that the expression vector pVAX/NA, of influenza A virus was successfully constructed. Western blot assay confirmed the expression of NA protein of influenza virus.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R392

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