本文選題：SARS冠狀病毒 + S蛋白　； 參考：《中國醫(yī)科大學(xué)》2006年碩士論文

【摘要】：目的 嚴(yán)重急性呼吸道綜合征(severe acute respiratory syndrome,SARS)是一種新型急性傳染性疾病,它的高死亡率嚴(yán)重威脅著人類的健康,在2004年仍有病例散發(fā)。其病原體是一種新的冠狀病毒(SARS Coronavirus,SARS-CoV),來源于野生動物,SARS-CoV是單股正鏈RNA病毒,基因組全長約29.7kb,纖突蛋白(S)、囊膜蛋白(E)、基質(zhì)蛋白(M)、及核衣殼蛋白(N)是它的主要結(jié)構(gòu)蛋白。其中S蛋白是SARS-CoV主要的膜結(jié)合蛋白,為Ⅰ型跨膜糖蛋白,在致病過程中S蛋白是主要的毒力因子,在病毒吸附細(xì)胞受體、細(xì)胞融合及誘導(dǎo)中和抗體產(chǎn)生過程中發(fā)揮關(guān)鍵作用。為了預(yù)防SARS的再次流行,人們迫切需要一種安全有效的疫苗早日研制成功。目前有許多候選疫苗進(jìn)入臨床前研究,例如:滅活疫苗,DNA疫苗,多肽疫苗等。我們的實驗是通過網(wǎng)絡(luò)數(shù)據(jù)庫結(jié)合生物信息學(xué)軟件分析的方法,確定S蛋白中可能在SARS-CoV感染過程中起重要作用并具有較好抗原性參數(shù)的結(jié)構(gòu)區(qū)域作為候選抗原表位,人工合成嵌合基因,體外表達(dá)后免疫家兔,為抗SARS基因工程疫苗的深入研究提供實驗依據(jù)。 方法 1.抗原表位預(yù)測 運(yùn)用生物信息學(xué)對SARS-CoV S蛋白的二級結(jié)構(gòu),親水性,穿膜螺旋,N-糖基化位點,氨基酸序列的親疏水性,表面可及性,分子剛性及抗原性參數(shù)進(jìn)行預(yù)測分析,設(shè)計抗原表位多肽。 2.基因合成和測序鑒定 設(shè)計串聯(lián)多表位的嵌合基因,命名為CAG,全長1274 bp由大連TaKaRa公司合成及測序驗證。 3.CAG嵌合基因的擴(kuò)增
[Abstract]:Purpose Severe acute respiratory syndrome (SARS) is a new type of acute infectious disease. Its high mortality is a serious threat to human health. The pathogen of SARS-CoV is a new coronavirus, SARS-CoV. SARS-CoV is a single-strand positive strand RNA virus. Its genome length is about 29.7 kb. The main structural proteins of SARS-CoV are the envelope protein, matrix protein, nucleocapsid protein and nucleocapsid protein. Among them, S protein is the main membrane binding protein of SARS-CoV, which is type I transmembrane glycoprotein. S protein is the main virulence factor in the pathogenesis of SARS-CoV. It plays a key role in the process of virus adsorbing cell receptor, cell fusion and inducing neutralizing antibody. In order to prevent the re-prevalence of SARS, it is urgent to develop a safe and effective vaccine as soon as possible. At present, there are many candidate vaccines, such as inactivated DNA vaccine, polypeptide vaccine and so on. Through the method of network database and bioinformatics software analysis, we identified the structural regions of S protein that may play an important role in the process of SARS-CoV infection and have good antigenicity parameters as candidate epitopes. The chimeric gene was synthesized and expressed in vitro to immunize rabbits. It provides experimental basis for further study of anti SARS gene engineering vaccine. Method 1. Antigen epitope prediction The secondary structure, hydrophilicity, N-glycosylation site of SARS-CoV S protein, hydrophobicity, surface accessibility, molecular rigidity and antigenicity parameters of amino acid sequence were predicted and analyzed by bioinformatics, and antigen epitope peptides were designed. 2. Gene synthesis and sequencing A chimeric gene named CAG was designed and its length of 1274 BP was synthesized and sequenced by Dalian TaKaRa Company. Amplification of 3.CAG Chimeric Gene
【學(xué)位授予單位】：中國醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2006
【分類號】：R392

【參考文獻(xiàn)】

相關(guān)期刊論文 前6條

1 廣東省防治非典型肺炎科技攻關(guān)專題組,徐安龍;SARS相關(guān)冠狀病毒基因工程重組蛋白質(zhì)疫苗的研制[J];廣東醫(yī)學(xué);2003年S1期

2 劉國棟;傳染性非典型肺炎病毒和疫苗的研究[J];中國國境衛(wèi)生檢疫雜志;2003年05期

3 董關(guān)木,王軍志;SARS疫苗的研究進(jìn)展[J];中國生物制品學(xué)雜志;2004年01期

4 張建中;傳染性非典型性肺炎的病原學(xué)和病理學(xué)研究現(xiàn)狀[J];實用醫(yī)學(xué)雜志;2003年07期

5 董學(xué)員,楊小昂,李燕,徐國賓,馮珍如,王月丹,陳慰峰;SARS病毒S2基因的克隆、表達(dá)、純化及其免疫學(xué)特性研究[J];細(xì)胞與分子免疫學(xué)雜志;2004年03期

6 趙雨杰,何群,馬佳明,王彤,呂昌龍,何欽成;SARS冠狀病毒基因組及其編碼蛋白生物信息學(xué)分析[J];中國醫(yī)科大學(xué)學(xué)報;2003年03期

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