[Expression, purification and identification of recombinant SARS coronavirus membrane protein].
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.
Zhang
Xiao-Li
XL
Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, the Chinese Academy of Sciences, Shanghai 200032, China.
Wang
Jing-Ru
JR
Zhang
Yan
Y
Chen
Mao-Lin
ML
Zhang
Wei
W
Yang
Sheng
S
Jiang
Wei-Hong
WH
D000595:Amino Acid Sequence; D015153:Blotting, Western; D002352:Carrier Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D000086862:Coronavirus M Proteins; D015967:Gene Expression Regulation, Viral; D058619:Maltose-Binding Proteins; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D045473:Severe acute respiratory syndrome-related coronavirus / Q000235:genetics* / Q000378:metabolism; D020539:Sequence Analysis, Protein; D014763:Viral Matrix Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
D002352:Carrier Proteins; D000086862:Coronavirus M Proteins; C000706237:M protein, SARS-CoV; D058619:Maltose-Binding Proteins; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D014763:Viral Matrix Proteins