SARS冠状病毒M基因在大肠杆菌中的表达及其免疫学特性初步研究

【目的】在大肠杆菌中表达SARS冠状病毒包膜(M)蛋白与谷胱甘肽S-转移酶(GST)融合蛋白,并对其进行血清学鉴定。【方法】克隆编码SARS冠状病毒M蛋白的基因,并在原核系统中表达GST-M融合蛋白,用Westernblot和酶联免疫吸附试验(ELISA)分析GST-M融合蛋白。【结果】10份SARS患者恢复期血清均能识别M-GST融合蛋白,并在Mr=52000附近出现特异性结合带;而10份正常人血清不与M-GST融合蛋白起反应。【结论】本研究获得了SARS冠状病毒GST-M融合蛋白,它可与SARS患者的恢复期血清产生特异性的结合反应,为研究SARS冠状病毒感染宿主细胞的过程和制备重组疫苗提供了条件。

Expression of SARS Coronavirus M Gene in E.Coli and Its Immunological Characteristics

To express membrane protein (M) of SARS coronavirus (SARS-CoV) fused with glutathione S-transferase (GST) and then detect its reactivity in the sera from convalescent SARS patients. The GST-M fusion protein was expressed in E.Coli, and then detected by Western blot and ELISA with 10 serum samples of convalescent SARS patients and 10 serum samples of healthy donors. According to the SDS-PAGE and ELISA analysis, the relative molecular mass (Mr) of the GST-M fusion protein is about 52 000. The fusion protein could react with all the sera from convalescent SARS patients, but not with the sera from healthy donors. Conclusion] The GST-M fusion protein provides a basis for the research on its role in the course of SARS coronavirus infection of host cells and preparation of recombinant vaccine against SARS coronavirus.