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

Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai). 2003 Dec;35(12):1140-4.
[Article in Chinese]

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Blotting, Western
  • Carrier Proteins / genetics
  • Carrier Proteins / isolation & purification
  • Carrier Proteins / metabolism
  • Coronavirus M Proteins
  • Gene Expression Regulation, Viral
  • Maltose-Binding Proteins
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / isolation & purification
  • Recombinant Fusion Proteins / metabolism
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism
  • Sequence Analysis, Protein
  • Severe acute respiratory syndrome-related coronavirus / genetics*
  • Severe acute respiratory syndrome-related coronavirus / metabolism
  • Viral Matrix Proteins / chemistry
  • Viral Matrix Proteins / genetics*
  • Viral Matrix Proteins / metabolism

Substances

  • Carrier Proteins
  • Coronavirus M Proteins
  • M protein, SARS-CoV
  • Maltose-Binding Proteins
  • Recombinant Fusion Proteins
  • Recombinant Proteins
  • Viral Matrix Proteins