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【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。
【Objective】 SARS coronavirus envelope (M) protein and glutathione S-transferase (GST) fusion protein were expressed in Escherichia coli and identified by serological method. 【Method】 The gene encoding the M protein of SARS coronavirus was cloned and the GST-M fusion protein was expressed in prokaryotic system. The GST-M fusion protein was analyzed by Western blot and enzyme-linked immunosorbent assay (ELISA). 【Results】 All the 10 SARS patients could recognize M-GST fusion protein in the serum of convalescent phase and showed specific binding band around Mr = 52000. However, 10 normal human serum did not react with M-GST fusion protein. 【Conclusion】 SARS-CoV GST-M fusion protein was obtained in this study. It can produce specific binding reaction with convalescent SARS patients. It provides the conditions for the study of SARS-CoV-infected host cells and preparation of recombinant vaccines.