两种人肠道病毒病毒样颗粒的制备与鉴定

发布时间：2018-01-11 04:26

本文关键词：两种人肠道病毒病毒样颗粒的制备与鉴定　出处：《北京工业大学》2012年硕士论文　论文类型：学位论文

【摘要】：肠道病毒属（enterovirus，EV）属于小RNA病毒科，，包括柯萨奇病毒（coxsackievirus，Cox）、新型肠道病毒（EV68-102）、脊髓灰质炎病毒（poliovirus，PV）和埃柯病毒（Echovirus）。其中，EV72目前被重命名为甲肝病毒(hepatitis virus A，HAV)。甲肝病毒是引起以肝脏损害为主的肠道传染病。我国是甲型肝炎的高发流行区，政府对甲肝预防亦是高度重视。目前，在我国应用的甲肝预防性疫苗有减毒活疫苗和灭活疫苗。这些疫苗的推广和应用对控制甲肝的传播与流行做出了重要贡献。CoxA16是引起患者手足口病（hand footmouth disease，HFMD）的主要病原体之一，同时该病毒还能引起疱疹性咽峡炎、脑膜脑炎等严重疾病，威胁了幼儿的生命健康。据报导，2012年第一季度北京市报告手足口病例比去年同期上升了216.67%，并且病原学监测中以CoxA16为主。目前，还没有针对CoxA16感染的特效药或疫苗上市。因此，依据当前HAV和CoxA16均没有基因工程重组疫苗的现状，本论文针对HAV和CoxA16开展基于病毒样颗粒的基因工程疫苗的探索研究。本论文选用了两种表达系统来制备HAV和CoxA16病毒样颗粒（virus-likeparticle，VLP）。第一，选择昆虫杆状病毒表达系统来表达HAV的VLP。将HAV的P1和3ABC基因重组到杆状病毒中并在昆虫细胞中共表达，电镜观察在昆虫细胞中可产生HAV VLP。第二，选择了酿酒酵母表达系统发酵表达CoxA16的VLP。将CoxA16的P1和3CD蛋白在酿酒酵母中共同表达，电镜观察可产生CoxA16的VLP。本论文得到了HAV和CoxA16的VLP。为研究预防甲肝病毒和科萨奇A16病毒感染的基因工程疫苗进行有益的探索。
[Abstract]:Enteroviruses belong to the small RNA virus family, including CoxsackievirusCox (CoxsackievirusCox.coxsackievirusCox.coxsackievirus. cox.coxsackievirus). New enterovirus EV68-102, poliovirusPVV) and Eco virus Echovirus. among them. EV72 has been renamed hepatitis virus A. Hepatitis A virus (HAV) is an enteric infectious disease mainly caused by liver damage. China is a high prevalence area of hepatitis A. the government also attaches great importance to the prevention of hepatitis A. at present. Live attenuated vaccines and inactivated vaccines are used in China. The promotion and application of these vaccines have made an important contribution to the control of the spread and epidemic of hepatitis A. CoxA16 is a cause of hand, foot and mouth disease (HFMD) in patients. Hand footmouth disease. HFMD is one of the main pathogens, and the virus can also cause herpes isthmus, meningoencephalitis and other serious diseases, threatening the life and health of young children. In in the first quarter of 2012, Beijing reported HFMD cases increased by 216.67 compared with the same period last year, and CoxA16 is the main etiological surveillance. There is no specific drug or vaccine for CoxA16 infection. Therefore, there is no current status of genetically engineered recombinant vaccine based on HAV and CoxA16. In this paper, the genetic engineering vaccine based on virus-like particles was developed for HAV and CoxA16. In this paper, two expression systems were selected to prepare virus-like particles of HAV and CoxA16 virus. Insect baculovirus expression system was selected to express HAV. The P1 and 3ABC genes of HAV were recombined into baculovirus and co-expressed in insect cells. Electron microscopic observation showed that HAV VLP could be produced in insect cells. The yeast expression system was selected to ferment and express CoxA16. The P1 and 3CD proteins of CoxA16 were co-expressed in Saccharomyces cerevisiae. Electron microscope was used to observe the formation of CoxA16. In this paper, HAV and CoxA16 VLPs have been obtained, which are useful for the study of genetic engineering vaccine against hepatitis A virus and Kosachi A16 virus infection.
【学位授予单位】：北京工业大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R373

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