兩種人腸道病毒病毒樣顆粒的制備與鑒定
本文關(guān)鍵詞:兩種人腸道病毒病毒樣顆粒的制備與鑒定 出處:《北京工業(yè)大學(xué)》2012年碩士論文 論文類型:學(xué)位論文
更多相關(guān)文章: 疫苗 病毒樣顆粒 甲型肝炎病毒 柯薩奇A16病毒 手足口病
【摘要】:腸道病毒屬(enterovirus,EV)屬于小RNA病毒科,,包括柯薩奇病毒(coxsackievirus,Cox)、新型腸道病毒(EV68-102)、脊髓灰質(zhì)炎病毒(poliovirus,PV)和�?虏《荆‥chovirus)。其中,EV72目前被重命名為甲肝病毒(hepatitis virus A,HAV)。甲肝病毒是引起以肝臟損害為主的腸道傳染病。我國是甲型肝炎的高發(fā)流行區(qū),政府對甲肝預(yù)防亦是高度重視。目前,在我國應(yīng)用的甲肝預(yù)防性疫苗有減毒活疫苗和滅活疫苗。這些疫苗的推廣和應(yīng)用對控制甲肝的傳播與流行做出了重要貢獻(xiàn)。CoxA16是引起患者手足口�。╤and footmouth disease,HFMD)的主要病原體之一,同時(shí)該病毒還能引起皰疹性咽峽炎、腦膜腦炎等嚴(yán)重疾病,威脅了幼兒的生命健康。據(jù)報(bào)導(dǎo),2012年第一季度北京市報(bào)告手足口病例比去年同期上升了216.67%,并且病原學(xué)監(jiān)測中以CoxA16為主。目前,還沒有針對CoxA16感染的特效藥或疫苗上市。因此,依據(jù)當(dāng)前HAV和CoxA16均沒有基因工程重組疫苗的現(xiàn)狀,本論文針對HAV和CoxA16開展基于病毒樣顆粒的基因工程疫苗的探索研究。 本論文選用了兩種表達(dá)系統(tǒng)來制備HAV和CoxA16病毒樣顆粒(virus-likeparticle,VLP)。第一,選擇昆蟲桿狀病毒表達(dá)系統(tǒng)來表達(dá)HAV的VLP。將HAV的P1和3ABC基因重組到桿狀病毒中并在昆蟲細(xì)胞中共表達(dá),電鏡觀察在昆蟲細(xì)胞中可產(chǎn)生HAV VLP。第二,選擇了釀酒酵母表達(dá)系統(tǒng)發(fā)酵表達(dá)CoxA16的VLP。將CoxA16的P1和3CD蛋白在釀酒酵母中共同表達(dá),電鏡觀察可產(chǎn)生CoxA16的VLP。 本論文得到了HAV和CoxA16的VLP。為研究預(yù)防甲肝病毒和科薩奇A16病毒感染的基因工程疫苗進(jìn)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.
【學(xué)位授予單位】:北京工業(yè)大學(xué)
【學(xué)位級別】:碩士
【學(xué)位授予年份】:2012
【分類號】:R373
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