[Expression, purification and identification of recombinant SARS coronavirus membrane protein].
Identifieur interne : 003045 ( PubMed/Corpus ); précédent : 003044; suivant : 003046[Expression, purification and identification of recombinant SARS coronavirus membrane protein].
Auteurs : Xiao-Li Zhang ; Jing-Ru Wang ; Yan Zhang ; Mao-Lin Chen ; Wei Zhang ; Sheng Yang ; Wei-Hong JiangSource :
- Sheng wu hua xue yu sheng wu wu li xue bao Acta biochimica et biophysica Sinica [ 0582-9879 ] ; 2003.
English descriptors
- KwdEn :
- Amino Acid Sequence, Blotting, Western, Carrier Proteins (genetics), Carrier Proteins (isolation & purification), Carrier Proteins (metabolism), 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), SARS Virus (genetics), SARS Virus (metabolism), Sequence Analysis, Protein, Viral Matrix Proteins (chemistry), Viral Matrix Proteins (genetics), Viral Matrix Proteins (metabolism).
- MESH :
- chemical , chemistry : Viral Matrix Proteins.
- chemical , genetics : Carrier Proteins, Recombinant Fusion Proteins, Viral Matrix Proteins.
- chemical , isolation & purification : Carrier Proteins, Recombinant Fusion Proteins, Recombinant Proteins.
- chemical , metabolism : Carrier Proteins, Recombinant Fusion Proteins, Recombinant Proteins, Viral Matrix Proteins.
- genetics : SARS Virus.
- metabolism : SARS Virus.
- Amino Acid Sequence, Blotting, Western, Gene Expression Regulation, Viral, Maltose-Binding Proteins, Sequence Analysis, Protein.
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.
PubMed: 14673508
Links to Exploration step
pubmed:14673508Le document en format XML
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<author><name sortKey="Zhang, Xiao Li" sort="Zhang, Xiao Li" uniqKey="Zhang X" first="Xiao-Li" last="Zhang">Xiao-Li Zhang</name>
<affiliation><nlm:affiliation>Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, the Chinese Academy of Sciences, Shanghai 200032, China.</nlm:affiliation>
</affiliation>
</author>
<author><name sortKey="Wang, Jing Ru" sort="Wang, Jing Ru" uniqKey="Wang J" first="Jing-Ru" last="Wang">Jing-Ru Wang</name>
</author>
<author><name sortKey="Zhang, Yan" sort="Zhang, Yan" uniqKey="Zhang Y" first="Yan" last="Zhang">Yan Zhang</name>
</author>
<author><name sortKey="Chen, Mao Lin" sort="Chen, Mao Lin" uniqKey="Chen M" first="Mao-Lin" last="Chen">Mao-Lin Chen</name>
</author>
<author><name sortKey="Zhang, Wei" sort="Zhang, Wei" uniqKey="Zhang W" first="Wei" last="Zhang">Wei Zhang</name>
</author>
<author><name sortKey="Yang, Sheng" sort="Yang, Sheng" uniqKey="Yang S" first="Sheng" last="Yang">Sheng Yang</name>
</author>
<author><name sortKey="Jiang, Wei Hong" sort="Jiang, Wei Hong" uniqKey="Jiang W" first="Wei-Hong" last="Jiang">Wei-Hong Jiang</name>
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<author><name sortKey="Zhang, Xiao Li" sort="Zhang, Xiao Li" uniqKey="Zhang X" first="Xiao-Li" last="Zhang">Xiao-Li Zhang</name>
<affiliation><nlm:affiliation>Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, the Chinese Academy of Sciences, Shanghai 200032, China.</nlm:affiliation>
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<author><name sortKey="Wang, Jing Ru" sort="Wang, Jing Ru" uniqKey="Wang J" first="Jing-Ru" last="Wang">Jing-Ru Wang</name>
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<author><name sortKey="Zhang, Yan" sort="Zhang, Yan" uniqKey="Zhang Y" first="Yan" last="Zhang">Yan Zhang</name>
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<author><name sortKey="Chen, Mao Lin" sort="Chen, Mao Lin" uniqKey="Chen M" first="Mao-Lin" last="Chen">Mao-Lin Chen</name>
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<author><name sortKey="Zhang, Wei" sort="Zhang, Wei" uniqKey="Zhang W" first="Wei" last="Zhang">Wei Zhang</name>
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<author><name sortKey="Yang, Sheng" sort="Yang, Sheng" uniqKey="Yang S" first="Sheng" last="Yang">Sheng Yang</name>
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<series><title level="j">Sheng wu hua xue yu sheng wu wu li xue bao Acta biochimica et biophysica Sinica</title>
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<term>Blotting, Western</term>
<term>Carrier Proteins (genetics)</term>
<term>Carrier Proteins (isolation & purification)</term>
<term>Carrier Proteins (metabolism)</term>
<term>Gene Expression Regulation, Viral</term>
<term>Maltose-Binding Proteins</term>
<term>Recombinant Fusion Proteins (genetics)</term>
<term>Recombinant Fusion Proteins (isolation & purification)</term>
<term>Recombinant Fusion Proteins (metabolism)</term>
<term>Recombinant Proteins (isolation & purification)</term>
<term>Recombinant Proteins (metabolism)</term>
<term>SARS Virus (genetics)</term>
<term>SARS Virus (metabolism)</term>
<term>Sequence Analysis, Protein</term>
<term>Viral Matrix Proteins (chemistry)</term>
<term>Viral Matrix Proteins (genetics)</term>
<term>Viral Matrix Proteins (metabolism)</term>
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<keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Viral Matrix Proteins</term>
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<keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Carrier Proteins</term>
<term>Recombinant Fusion Proteins</term>
<term>Recombinant Proteins</term>
<term>Viral Matrix Proteins</term>
</keywords>
<keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>SARS Virus</term>
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<keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>SARS Virus</term>
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<keywords scheme="MESH" xml:lang="en"><term>Amino Acid Sequence</term>
<term>Blotting, Western</term>
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<front><div type="abstract" xml:lang="en">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.</div>
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<Title>Sheng wu hua xue yu sheng wu wu li xue bao Acta biochimica et biophysica Sinica</Title>
<ISOAbbreviation>Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao</ISOAbbreviation>
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<ArticleTitle>[Expression, purification and identification of recombinant SARS coronavirus membrane protein].</ArticleTitle>
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<Abstract><AbstractText>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.</AbstractText>
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