【摘要】 以大肠杆菌表达载体pET22b为载体,直接表达SARS病毒S蛋白425～569及894～1033等2片段。表达所获得的包涵体形式蛋白经尿素溶解后分别经过2次离子交换层析,获得初步纯化。在酸性和低尿素浓度环境中,2种S蛋白片段极易沉淀。Western印迹鉴定显示其与抗SARS病毒血清呈阳性反应。获得的纯化蛋白可用于检验受体结合能力等研究。更多还原

【Abstract】 Severe acute respiratory syndrome(SARS) is an emerging infectious disease associated with a novel coronavirus and causing worldwide outbreaks. SARS coronavirus(SARS-CoV) is an enveloped RNA virus, which contains several structural proteins. Among these proteins, spike(S) protein is responsible for binding to specific cellular receptors and is a major antigenic determinant, which induces neutralizing antibody. The N-terminus 425～569 and 894～1033 residuals of S glycoprotein of SARS-CoV have been expressed in Escherichia coli using pET22b expression vector. After lactose induction the truncated spike protein were expressed in inclusion body form. The positive reaction between purified truncated protein and anti-SARS sera is clear in Western blot. And we found difficult to renatured the purified protein in urea using common refolding system. The purified protein can be used to study receptor-binding ability.更多还原