SARS冠状病毒S2基因原核表达及免疫学特性

Prokaryotic expression of SARS-CoV S2 gene and preliminary study on its immune characteristics

目的研究严重急性呼吸道综合征(SARS)冠状病毒S2蛋白表达并对其免疫学特性。方法克隆SARS冠状病毒S2基因,并在大肠埃希菌中表达谷胱甘肽硫转移酶(GST-S2)融合蛋白。通过免疫印迹(Westernblot)和酶联免疫吸附试验(ELISA)方法鉴定表达GST-S2融合蛋白的活性。结果通过原核系统表达的GST-S2融合蛋白可与谷胱甘肽硫转移酶(GST)多克隆抗体结合,并在85千道尔顿(KD)处出现特异性结合条带。GST-S2蛋白能与SARS患者恢复期血清反应,而不与正常人血清反应。结论本项研究获得了SARS冠状病毒GST-S2融合蛋白,它可与GST多克隆抗体特异性结合,并与SARS患者恢复期血清发生特异性反应。

Objective To express the SARS-CoV S2 protein in E coli and to study on its immune characteristics preliminarily. Methods SARS-CoV S2 gene was cloned and the GST-S2 fusion protein was expressed in E coli. The activity of the expressed GST-S2 fusion protein was identified by Western blot and enzyme linked immunosorbent assay （ELISA）. Results Prokaryotie expressed GST-S2 fusion protein could bind the GST polyclone antibody and an 85KD immune band could be detected. Expressed GST-S2 protein bad response to convalescent serum from SARS patients, but not to control serum. Conclusion The obtained GST-S2 protein could be provided for the study on the infection process of host cells by SARS-CoV and the preparation of the recombination vaccine.