本文選題：SARS + S蛋白　； 參考：《蘇州大學(xué)》2005年碩士論文

【摘要】：目的:用分子對接技術(shù)預(yù)測HIV融合抑制劑是否能對SARS起作用;原核表達SARS S蛋白S2結(jié)構(gòu)域,制備抗S2的多克隆抗體。方法:搜尋HIV gp41融合抑制劑,用DOCK軟件分析其與SARS S2結(jié)構(gòu)域HR1三聚體晶體結(jié)構(gòu)的親和力;用PCR技術(shù)得到S2結(jié)構(gòu)域編碼序列,亞克隆至原核表達載體pGEX-5X-3,轉(zhuǎn)化大腸桿菌BL21,IPTG誘導(dǎo)表達融合蛋白。所得S2融合蛋白經(jīng)電泳后KCl染色割膠,免疫家兔制備多抗,分離血清并用瓊脂糖雙向擴散實驗測定抗體效價。結(jié)果:預(yù)測了23個小分子,ADS-J1、ADS-J2、XTT Formazan等幾個小分子得分較高,提示有親和力。構(gòu)建了pGEX-5X-3-S2原核表達載體,SDS-PAGE電泳顯示誘導(dǎo)出84.2KD融合蛋白。融合蛋白免疫家兔,瓊脂糖雙向擴散實驗證實血清有一定的抗體效價。結(jié)論:發(fā)現(xiàn)抗HIV的融合抑制劑確能對SARS起作用。成功表達了S2結(jié)構(gòu)域,并制備了抗S2結(jié)構(gòu)域的多克隆抗體。
[Abstract]:Objective: to predict whether HIV fusion inhibitor can act on SARS by molecular docking technique, and to express S 2 domain of SARS protein in prokaryotic expression and to prepare polyclonal antibody against S 2. Methods: gp41 fusion inhibitor was searched, and its affinity to HR1 trimer crystal structure of S2 domain was analyzed by dock software, and the coding sequence of S2 domain was obtained by PCR. Subcloned into prokaryotic expression vector pGEX-5X-3 and transformed into E. coli BL21 and IPTG to express fusion protein. The S 2 fusion protein was stained with KCl to prepare polyclonal antibodies. The antibody titers were determined by two dimensional agarose diffusion assay. Results: the scores of 23 small molecules ADS-J1 and ADS-J2 + XTT Formazan were higher, indicating that they had affinity. A prokaryotic expression vector pGEX-5X-3-S2 was constructed. SDS-PAGE showed that 84.2 KD fusion protein was induced by SDS-PAGE. Rabbits were immunized with the fusion protein, and the agarose diffusion assay confirmed that the serum had a certain antibody titer. Conclusion: the fusion inhibitor of anti-HIV can play an important role in SARS. The S2 domain was successfully expressed and the polyclonal antibody against S2 domain was prepared.
【學(xué)位授予單位】：蘇州大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2005
【分類號】：R373;R392

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本文編號：2075367