Expression, purification and sublocalization of SARS-CoV nucleocapsid protein in insect cells.
Identifieur interne : 005354 ( Main/Exploration ); précédent : 005353; suivant : 005355Expression, purification and sublocalization of SARS-CoV nucleocapsid protein in insect cells.
Auteurs : Ai-Xia Ren [République populaire de Chine] ; You-Hua Xie ; Yu-Ying Kong ; Guan-Zhen Yang ; Yao-Zhou Zhang ; Yuan Wang ; Xiang-Fu WuSource :
- Acta biochimica et biophysica Sinica [ 1672-9145 ] ; 2004.
Descripteurs français
- KwdFr :
- Animaux, Conformation des protéines, Cytoplasme (), Escherichia coli (métabolisme), Humains, Lignée cellulaire, Microscopie confocale, Microscopie de fluorescence, Plasmides (métabolisme), Protéines nucléocapside (), Protéines nucléocapside (isolement et purification), Protéines nucléocapside (métabolisme), Protéines recombinantes (), Protéines recombinantes (métabolisme), Technique de Western, Virus du SRAS (métabolisme), Électrophorèse sur gel de polyacrylamide.
- MESH :
- isolement et purification : Protéines nucléocapside.
- métabolisme : Escherichia coli, Plasmides, Protéines nucléocapside, Protéines recombinantes, Virus du SRAS.
- Animaux, Conformation des protéines, Cytoplasme, Humains, Lignée cellulaire, Microscopie confocale, Microscopie de fluorescence, Protéines nucléocapside, Protéines recombinantes, Technique de Western, Électrophorèse sur gel de polyacrylamide.
English descriptors
- KwdEn :
- Animals, Blotting, Western, Cell Line, Cytoplasm (chemistry), Electrophoresis, Polyacrylamide Gel, Escherichia coli (metabolism), Humans, Insecta, Microscopy, Confocal, Microscopy, Fluorescence, Nucleocapsid Proteins (chemistry), Nucleocapsid Proteins (isolation & purification), Nucleocapsid Proteins (metabolism), Plasmids (metabolism), Protein Conformation, Recombinant Proteins (chemistry), Recombinant Proteins (metabolism), SARS Virus (metabolism).
- MESH :
- chemical , chemistry : Nucleocapsid Proteins, Recombinant Proteins.
- chemistry : Cytoplasm.
- chemical , isolation & purification : Nucleocapsid Proteins.
- metabolism : Escherichia coli, Nucleocapsid Proteins, Plasmids, Recombinant Proteins, SARS Virus.
- Animals, Blotting, Western, Cell Line, Electrophoresis, Polyacrylamide Gel, Humans, Insecta, Microscopy, Confocal, Microscopy, Fluorescence, Protein Conformation.
Abstract
The causative agent of severe acute respiratory syndrome (SARS) is a previously unidentified coronavirus, SARS-CoV. The nucleocapsid (N) protein of SARS-CoV is a major viral protein recognized by acute and early convalescent sera from SARS patients. To facilitate the studies on the function and structure of the N protein, this report describe the expression and purification of recombinant SARS-CoV N protein using the baculovirus expression system. Recombinant hexa-histidine-tagged N protein with a molecular mass of 47 kD was produced in insect cells. Recombinant N protein was purified to near homogeneity by Ni2+-NTA affinity chromatography. In addition, we examined the subcellular localization of the N protein by confocal microscopy in Trichoplusia ni BT1 Tn 5B1-4 cells infected with recombinant baculovirus. The N protein was found localized in the cytoplasm as well as in the nucleolus. The purified recombinant N protein can be used in further functional study of SARS-CoV.
DOI: 10.1093/abbs/36.11.754
PubMed: 15514849
Affiliations:
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Le document en format XML
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uniqKey="Kong Y" first="Yu-Ying" last="Kong">Yu-Ying Kong</name></author><author><name sortKey="Yang, Guan Zhen" sort="Yang, Guan Zhen" uniqKey="Yang G" first="Guan-Zhen" last="Yang">Guan-Zhen Yang</name></author><author><name sortKey="Zhang, Yao Zhou" sort="Zhang, Yao Zhou" uniqKey="Zhang Y" first="Yao-Zhou" last="Zhang">Yao-Zhou Zhang</name></author><author><name sortKey="Wang, Yuan" sort="Wang, Yuan" uniqKey="Wang Y" first="Yuan" last="Wang">Yuan Wang</name></author><author><name sortKey="Wu, Xiang Fu" sort="Wu, Xiang Fu" uniqKey="Wu X" first="Xiang-Fu" last="Wu">Xiang-Fu Wu</name></author></analytic><series><title level="j">Acta biochimica et biophysica Sinica</title><idno type="ISSN">1672-9145</idno><imprint><date when="2004" type="published">2004</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term><term>Blotting, Western</term><term>Cell Line</term><term>Cytoplasm 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purification)</term><term>Protéines nucléocapside (métabolisme)</term><term>Protéines recombinantes ()</term><term>Protéines recombinantes (métabolisme)</term><term>Technique de Western</term><term>Virus du SRAS (métabolisme)</term><term>Électrophorèse sur gel de polyacrylamide</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Nucleocapsid Proteins</term><term>Recombinant Proteins</term></keywords><keywords scheme="MESH" qualifier="chemistry" xml:lang="en"><term>Cytoplasm</term></keywords><keywords scheme="MESH" type="chemical" qualifier="isolation & purification" xml:lang="en"><term>Nucleocapsid Proteins</term></keywords><keywords scheme="MESH" qualifier="isolement et purification" xml:lang="fr"><term>Protéines nucléocapside</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Escherichia coli</term><term>Nucleocapsid Proteins</term><term>Plasmids</term><term>Recombinant Proteins</term><term>SARS 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polyacrylamide</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The causative agent of severe acute respiratory syndrome (SARS) is a previously unidentified coronavirus, SARS-CoV. The nucleocapsid (N) protein of SARS-CoV is a major viral protein recognized by acute and early convalescent sera from SARS patients. To facilitate the studies on the function and structure of the N protein, this report describe the expression and purification of recombinant SARS-CoV N protein using the baculovirus expression system. Recombinant hexa-histidine-tagged N protein with a molecular mass of 47 kD was produced in insect cells. Recombinant N protein was purified to near homogeneity by Ni2+-NTA affinity chromatography. In addition, we examined the subcellular localization of the N protein by confocal microscopy in Trichoplusia ni BT1 Tn 5B1-4 cells infected with recombinant baculovirus. The N protein was found localized in the cytoplasm as well as in the nucleolus. The purified recombinant N protein can be used in further functional study of SARS-CoV.</div></front></TEI><affiliations><list><country><li>République populaire de Chine</li></country><region><li>Zhejiang</li></region><settlement><li>Hangzhou</li></settlement><orgName><li>Université de Zhejiang</li></orgName></list><tree><noCountry><name sortKey="Kong, Yu Ying" sort="Kong, Yu Ying" uniqKey="Kong Y" first="Yu-Ying" last="Kong">Yu-Ying Kong</name><name sortKey="Wang, Yuan" sort="Wang, Yuan" uniqKey="Wang Y" first="Yuan" last="Wang">Yuan Wang</name><name sortKey="Wu, Xiang Fu" sort="Wu, Xiang Fu" uniqKey="Wu X" first="Xiang-Fu" last="Wu">Xiang-Fu Wu</name><name sortKey="Xie, You Hua" sort="Xie, You Hua" uniqKey="Xie Y" first="You-Hua" last="Xie">You-Hua Xie</name><name sortKey="Yang, Guan Zhen" sort="Yang, Guan Zhen" uniqKey="Yang G" first="Guan-Zhen" last="Yang">Guan-Zhen Yang</name><name sortKey="Zhang, Yao Zhou" sort="Zhang, Yao Zhou" uniqKey="Zhang Y" first="Yao-Zhou" last="Zhang">Yao-Zhou Zhang</name></noCountry><country name="République populaire de Chine"><region name="Zhejiang"><name sortKey="Ren, Ai Xia" sort="Ren, Ai Xia" uniqKey="Ren A" first="Ai-Xia" last="Ren">Ai-Xia Ren</name></region></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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