SARS病毒S1蛋白重组C端片段免疫效果的实验研究

為獲得純化的重組SARS病毒S1蛋白C端 ,研究其刺激機體產生針對SARS病毒免疫應答的規律和機制 ,將編碼SARS病毒S1蛋白C端 311個氨基酸殘基的基因克隆 ,并在原核表達系統中表達 ,純化獲得了的重組蛋白。利用SARS患者恢復期的血清 ,對純化的重組S1蛋白進行血清學分析。結果表明 ,本研究中克隆表達的重組蛋白序列與公布的SARS病毒S1蛋白C端的序列相同 ,其編碼的重組蛋白相對分子質量約為 5 90 0 0Mr。SARS患者恢復期的血清均與重組蛋白反應 ,在5 9 0 0 0Mr處形成特異性的反應條帶 ,而來自SARS流行前的正常人對照血清則不能與重組蛋白反應。在本研究中獲得的重組蛋白可以為研究SARS病毒識別宿主細胞受體的過程及其機制提供條件。To express the recombinant C-terminal fragment of SARS virus S1 protein and to study its role in SARS immune response. The gene encoding C-terminal 311 amino acid residuals of SARS virus S1 subunit was cloned and expressed in E.coli. After purification, the recombinant protein was identified from serum sample of patients recovered from SARS. The serum sample of health donor, which was collected befove outbreak of SARS, was used as negative control. Sequencing analysis confirmed that the recombinant protein has the same sequence of natural N-terminal of SARS virus S1 subunit. The recombinant S1 protein could react with the sample from recovered SARS patients, and showed a specific band at 59 000 Mr, but not the control sample in the result of Western blots. The recombinant N-terminal of SARS virus S1 subunit may provide us a good tool in the research of immune response to SARS virus in vivo and in vitro.