[Expression, purification and identification of recombinant SARS coronavirus membrane protein].

Xiao-Li Zhang, Jing-Ru Wang, Yan Zhang, Mao-Lin Chen, Wei Zhang, Sheng Yang, Wei-Hong Jiang
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Abstract

A novel coronavirus (SARS-coronavirus, SARS-CoV) was discovered as the pathogen of the severe acute respiratory syndrome (SARS). According to studies with other coronaviruses, the membrane protein (M protein) is the main structural protein and the recombinant M protein may be useful as an antigen for detecting antibodies against coronavirus and for preparing vaccine. In this work, the M protein of SARS-CoV was expressed in E. coli as fusion protein with maltose binding protein at N-terminus and MxeGyrA intein CBD at C-terminus. The recombinant protein was identified by Western blot and mass spectrometry. The soluble parts of the cell crude extract were then partially purified by MBP affinity chromatography. The purified protein will be used for the studies on M protein's structure and the development of diagnostic method of SARS.

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重组SARS冠状病毒膜蛋白的表达、纯化与鉴定
一种新型冠状病毒(SARS-冠状病毒,SARS- cov)被发现为严重急性呼吸综合征(SARS)的病原体。根据与其他冠状病毒的研究,膜蛋白(M蛋白)是其主要结构蛋白,重组M蛋白可作为检测冠状病毒抗体和制备疫苗的抗原。在本研究中,SARS-CoV的M蛋白以融合蛋白的形式在大肠杆菌中表达,融合蛋白在n端与麦芽糖结合蛋白(maltose binding protein)结合,在c端与MxeGyrA intin CBD结合。重组蛋白经Western blot和质谱鉴定。然后用MBP亲和层析对细胞粗提物的可溶性部分进行部分纯化。纯化后的蛋白将用于M蛋白的结构研究和SARS诊断方法的开发。
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