SARS病毒S蛋白片段的克隆表达和纯化  

The Expression and Purification of Truncated S Protein of SARS-CoV

作　　者：陈璟[1,2] 李彦英[3] 周育森[4] 李玉玲[3] 周长林[1] 方宏清[3] 陈惠鹏[3] 

机构地区：[1]中国药科大学生命科学与技术学院 [2]军事医学科学院生物工程研究所北京100071 [3]军事医学科学院生物工程研究所 [4]军事医学科学院微生物流行病研究所

出　　处：《生物技术通讯》2005年第3期249-251,共3页Letters in Biotechnology

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。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.

关 键 词：SARS病毒 S蛋白 纯化 克隆表达 片段 WESTERN印迹 离子交换层析 表达载体 大肠杆菌 尿素浓度 阳性反应 结合能力 体形 血清 受体 

分 类 号：Q78[生物学—分子生物学] TS941.724[轻工技术与工程—服装设计与工程]