SARS冠状病毒膜蛋白膜内区的表达、纯化及抗原性初步研究

Expression, purification and antigenicity of SARS-CoV membrane protein＇ s fragrant from inside membrane

目的对SARS冠状病毒膜蛋白膜内区基因片段进行克隆表达和表达产物的纯化复性，探讨该表达蛋白的抗原特性。方法克隆SARS冠状病毒GD322株膜蛋白膜内区，在原核表达系统中表达His-融合蛋白，采用Western blot和酶联免疫吸附试验（ELISA）分析His-融合蛋白抗原特性。结果7份SARS临床诊断患者血清中5份血清能与His-融合蛋白反应，2份不与之反应。兔抗OCA3、229E血清及20份健康人血清皆不与His-融合蛋白反应。His-融合蛋白免疫动物可产生多克隆抗体。结论本研究获得的His-融合蛋白可与SARS临床诊断患者血清特异性结合，可免疫动物获得特异性多抗；但不与健康人血清及兔抗OCA3、229E血清发生特异性结合。

Objective To test the antigenicity of SARS-CoV M protein. Methods After cloning SARS-CoV membrane protein＇s bases which locate inside the membrane, the His-fusion protein was expressed in E. coli and was analyzed. Results Among the 7 clinically diagnosed patients samples, 5 reacted with His-fusion protein; all of the 20 healthy persons＇ serum and rabbit anti-OC43 and 229E did not react with His-fusion protein; the animals immunized with His-fusion protein produced antibodies. Conclusion The His-fusion protein specially reacts with clinically diagnosed SAILS patients＇s serum and the animals immunized with His-fusion protein produce specific antibodies.