發(fā)布時間：2018-07-18 09:09

【摘要】：SARS (Severe Acute Respiratory Syndrome)是一種傳染性強、病死率高、嚴(yán)重危害人民生命健康的傳染病。世界衛(wèi)生組織于2003年4月6日宣布引起SARS的病原體是一種新的病毒,屬于冠狀病毒家族,稱為SARS相關(guān)冠狀病毒(SAPS-Associated coronavirus, SARS-CoV)。SARS-CoV表現(xiàn)出許多不同于冠狀病毒的特性,為冠狀病毒中一個新的分支。目前,病毒性疾病尚無特效治療方法,對于SARS也是一樣。所以,控制SARS的最好辦法是預(yù)防,而疫苗的開發(fā)則是建立有效防范措施的關(guān)鍵環(huán)節(jié)。DNA疫苗是繼病原體疫苗、亞單位疫苗之后的第三代疫苗,具有許多的優(yōu)越性。所以本研究選擇了SARS病毒DNA疫苗這一方向,以SARS-CoV主要結(jié)構(gòu)蛋白之一——M蛋白基因作為研究對象。構(gòu)建針對SARS-CoVM蛋白基因的重組核酸疫苗,觀察其免疫小鼠后機體的特異性免疫應(yīng)答的情況,探討其作為抗SARS-CoV疫苗的可能性。 本研究由兩部分組成: 第一部分為 SARS-CoV M蛋白基因重組核酸疫苗的構(gòu)建。從GenBank上獲取SARS病毒株SIN2774的M蛋白基因序列,使用引物設(shè)計軟件Primer Premier 5.0進(jìn)行引物設(shè)計,從中選擇最佳引物,并在引物5’端分別添加真核表達(dá)質(zhì)粒pcDNA3.1(+)多克隆位點中所包含的EcoRⅠ和Xho Ⅰ酶切位點及保護(hù)堿基,以從新加坡獲得的SIN2774 cDNA為模
[Abstract]:SARS (severe Acute Respiratory Syndrome) is a kind of infectious disease with high mortality and serious harm to people's life and health. The World Health Organization (WHO) announced on April 6, 2003 that the pathogen causing SARS is a new virus, belonging to the coronavirus family, called SARS-associated coronavirus (SARS-CoV). SARS-CoV shows many characteristics different from coronavirus. It is a new branch of coronavirus. At present, there is no special treatment for viral diseases, and the same is true for SARS. Therefore, the best way to control SARS is prevention, and the development of vaccine is the key link to establish effective preventive measures. DNA vaccine is the third generation vaccine after pathogen vaccine and subunit vaccine, which has many advantages. In this study, we chose the SARS-CoV DNA vaccine as the direction, and took M protein gene, one of the main structural proteins of SARS-CoV, as the research object. The recombinant nucleic acid vaccine against SARS-CoVM protein gene was constructed to observe the specific immune response of mice after immunization, and to explore the possibility of the recombinant nucleic acid vaccine as an anti-SARS-CoV vaccine. This study consists of two parts: the first part is the construction of SARS-CoV M gene recombinant nucleic acid vaccine. The M protein gene sequence of SARS virus strain SIN2774 was obtained from GenBank. Primer design software Primer Premier 5.0 was used to design the primers, and the best primers were selected. The eukaryotic expression plasmids pcDNA3.1 () polyclonal sites containing Ecor 鈪，

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