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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。
The Escherichia coli expression vector pET22b was used as a vector to directly express the 2 fragments of SARS virus S protein 425 ~ 569 and 894 ~ 1033. The expressed protein in inclusion body was dissolved in urea and subjected to 2 times of ion exchange chromatography to obtain preliminary purification. In acidic and low urea concentration environment, two kinds of S protein fragments easily precipitate. Western blotting showed that it was positive with anti-SARS virus serum. The purified protein can be used to test the binding ability of the receptor.