Expression, purification and sublocalization of SARS-CoV nucleocapsid protein in insect cells.
Identifieur interne : 002A74 ( PubMed/Corpus ); précédent : 002A73; suivant : 002A75Expression, purification and sublocalization of SARS-CoV nucleocapsid protein in insect cells.
Auteurs : Ai-Xia Ren ; 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.
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
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pubmed:15514849Le document en format XML
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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><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">15514849</PMID><DateCompleted><Year>2005</Year><Month>04</Month><Day>18</Day></DateCompleted><DateRevised><Year>2020</Year><Month>04</Month><Day>03</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Print">1672-9145</ISSN><JournalIssue CitedMedium="Print"><Volume>36</Volume><Issue>11</Issue><PubDate><Year>2004</Year><Month>Nov</Month></PubDate></JournalIssue><Title>Acta biochimica et biophysica Sinica</Title><ISOAbbreviation>Acta Biochim. Biophys. Sin. (Shanghai)</ISOAbbreviation></Journal><ArticleTitle>Expression, purification and sublocalization of SARS-CoV nucleocapsid protein in insect cells.</ArticleTitle><Pagination><MedlinePgn>754-8</MedlinePgn></Pagination><Abstract><AbstractText>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.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Ren</LastName><ForeName>Ai-Xia</ForeName><Initials>AX</Initials><AffiliationInfo><Affiliation>Institute of Biochemistry and Biotechnology, College of Life Science, Zhejiang University, Hangzhou 310029, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Xie</LastName><ForeName>You-Hua</ForeName><Initials>YH</Initials></Author><Author ValidYN="Y"><LastName>Kong</LastName><ForeName>Yu-Ying</ForeName><Initials>YY</Initials></Author><Author ValidYN="Y"><LastName>Yang</LastName><ForeName>Guan-Zhen</ForeName><Initials>GZ</Initials></Author><Author ValidYN="Y"><LastName>Zhang</LastName><ForeName>Yao-Zhou</ForeName><Initials>YZ</Initials></Author><Author ValidYN="Y"><LastName>Wang</LastName><ForeName>Yuan</ForeName><Initials>Y</Initials></Author><Author ValidYN="Y"><LastName>Wu</LastName><ForeName>Xiang-Fu</ForeName><Initials>XF</Initials></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType></PublicationTypeList></Article><MedlineJournalInfo><Country>China</Country><MedlineTA>Acta Biochim Biophys Sin (Shanghai)</MedlineTA><NlmUniqueID>101206716</NlmUniqueID><ISSNLinking>1672-9145</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D019590">Nucleocapsid Proteins</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D011994">Recombinant Proteins</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="C099602">nucleocapsid protein, Coronavirus</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D000818" MajorTopicYN="N">Animals</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D015153" MajorTopicYN="N">Blotting, Western</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D002460" MajorTopicYN="N">Cell Line</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D003593" MajorTopicYN="N">Cytoplasm</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="N">chemistry</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D004591" MajorTopicYN="N">Electrophoresis, Polyacrylamide Gel</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D004926" MajorTopicYN="N">Escherichia coli</DescriptorName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D006801" MajorTopicYN="N">Humans</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D007313" MajorTopicYN="N">Insecta</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D018613" MajorTopicYN="N">Microscopy, Confocal</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D008856" MajorTopicYN="N">Microscopy, Fluorescence</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D019590" MajorTopicYN="N">Nucleocapsid Proteins</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="Y">chemistry</QualifierName><QualifierName UI="Q000302" MajorTopicYN="Y">isolation & purification</QualifierName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D010957" MajorTopicYN="N">Plasmids</DescriptorName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D011487" MajorTopicYN="N">Protein Conformation</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D011994" MajorTopicYN="N">Recombinant Proteins</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="N">chemistry</QualifierName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D045473" MajorTopicYN="N">SARS Virus</DescriptorName><QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName></MeshHeading></MeshHeadingList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="pubmed"><Year>2004</Year><Month>10</Month><Day>30</Day><Hour>9</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2005</Year><Month>4</Month><Day>19</Day><Hour>9</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2004</Year><Month>10</Month><Day>30</Day><Hour>9</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">15514849</ArticleId><ArticleId IdType="doi">10.1093/abbs/36.11.754</ArticleId><ArticleId IdType="pmc">PMC7110235</ArticleId></ArticleIdList></PubmedData></pubmed></record>Pour manipuler ce document sous Unix (Dilib)
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