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Updated: Jun 21, 2026
Protocol for Recombinant RBD-based SARS Vaccines: Protein Preparation, Animal Vaccination and Neutralization Detection
Published on: May 2, 2011
[Expression, purification and identification of recombinant SARS coronavirus membrane protein]
Xiao-Li Zhang1, Jing-Ru Wang, Yan Zhang
1Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, the Chinese Academy of Sciences, Shanghai 200032, China.
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.
Insights
Researchers expressed and purified the SARS-CoV M protein in E. coli. This recombinant M protein is crucial for developing SARS diagnostic methods and potential vaccines.
Area of Science:
- Virology
- Molecular Biology
- Biochemistry
Background:
- Severe Acute Respiratory Syndrome (SARS) is caused by a novel coronavirus, SARS-CoV.
- The SARS-CoV membrane protein (M protein) is a major structural component.
- Recombinant M protein holds potential as an antigen for antibody detection and vaccine development.
Purpose of the Study:
- To express and purify the SARS-CoV M protein.
- To facilitate structural studies of the M protein.
- To develop a diagnostic method for SARS.
Main Methods:
- Expression of SARS-CoV M protein in E. coli as a fusion protein.
- Purification using MBP affinity chromatography.
- Identification via Western blot and mass spectrometry.
Main Results:
- Successfully expressed and identified the recombinant SARS-CoV M protein.
- Partially purified the soluble M protein from E. coli crude extract.
- The purified protein is suitable for further structural and diagnostic studies.
Conclusions:
- The recombinant SARS-CoV M protein was successfully produced and purified.
- This protein serves as a valuable tool for SARS research.
- Further studies will focus on M protein structure and SARS diagnostics.
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