SARS病毒S1蛋白重组C端片段免疫效果的实验研究

为获得纯化的重组SARS病毒S1蛋白C端 ,研究其刺激机体产生针对SARS病毒免疫应答的规律和机制 ,将编码SARS病毒S1蛋白C端 311个氨基酸残基的基因克隆 ,并在原核表达系统中表达 ,纯化获得了的重组蛋白。利用SARS患者恢复期的血清 ,对纯化的重组S1蛋白进行血清学分析。结果表明 ,本研究中克隆表达的重组蛋白序列与公布的SARS病毒S1蛋白C端的序列相同 ,其编码的重组蛋白相对分子质量约为 5 90 0 0Mr。SARS患者恢复期的血清均与重组蛋白反应 ,在5 9 0 0 0Mr处形成特异性的反应条带 ,而来自SARS流行前的正常人对照血清则不能与重组蛋白反应。在本研究中获得的重组蛋白可以为研究SARS病毒识别宿主细胞受体的过程及其机制提供条件。

Role of C-Terminal Fragment of SARS S1 Subunit in Anti-SARS Virus Immune Response

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.