High expression level of soluble SARS spike protein mediated by adenovirus in HEK293 cells.
Identifieur interne : 002302 ( PubMed/Corpus ); précédent : 002301; suivant : 002303High expression level of soluble SARS spike protein mediated by adenovirus in HEK293 cells.
Auteurs : Fei Zhong ; Zhen-Yu Zhong ; Shuang Liang ; Xiu-Jin LiSource :
- World journal of gastroenterology [ 1007-9327 ] ; 2006.
English descriptors
- KwdEn :
- Adenoviridae (genetics), Blotting, Western, Cell Line, Cell Survival, Gene Expression Regulation, Viral, Genetic Vectors, Humans, Membrane Glycoproteins (biosynthesis), Membrane Glycoproteins (genetics), Plasmids, Recombinant Proteins (analysis), SARS Virus, Spike Glycoprotein, Coronavirus, Viral Envelope Proteins (biosynthesis), Viral Envelope Proteins (genetics).
- MESH :
- chemical , analysis : Recombinant Proteins.
- chemical , biosynthesis : Membrane Glycoproteins, Viral Envelope Proteins.
- genetics : Adenoviridae, Membrane Glycoproteins, Viral Envelope Proteins.
- Blotting, Western, Cell Line, Cell Survival, Gene Expression Regulation, Viral, Genetic Vectors, Humans, Plasmids, SARS Virus, Spike Glycoprotein, Coronavirus.
Abstract
To develop a highly efficacious method for preparation of soluble SARS S-protein using adenovirus vector to meet the requirement for S-protein investigation.
DOI: 10.3748/wjg.v12.i9.1452
PubMed: 16552820
Links to Exploration step
pubmed:16552820Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">High expression level of soluble SARS spike protein mediated by adenovirus in HEK293 cells.</title><author><name sortKey="Zhong, Fei" sort="Zhong, Fei" uniqKey="Zhong F" first="Fei" last="Zhong">Fei Zhong</name><affiliation><nlm:affiliation>Department of Basic Veterinary Medicine, School of Animal Science and Technology, Hebei Agricultural University, Baoding 071001, Hebei Province, China. feizhong2000@yahoo.com</nlm:affiliation></affiliation></author><author><name sortKey="Zhong, Zhen Yu" sort="Zhong, Zhen Yu" uniqKey="Zhong Z" first="Zhen-Yu" last="Zhong">Zhen-Yu Zhong</name></author><author><name sortKey="Liang, Shuang" sort="Liang, Shuang" uniqKey="Liang S" first="Shuang" last="Liang">Shuang Liang</name></author><author><name sortKey="Li, Xiu Jin" sort="Li, Xiu Jin" uniqKey="Li X" first="Xiu-Jin" last="Li">Xiu-Jin Li</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2006">2006</date><idno type="RBID">pubmed:16552820</idno><idno type="pmid">16552820</idno><idno type="doi">10.3748/wjg.v12.i9.1452</idno><idno type="wicri:Area/PubMed/Corpus">002302</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">002302</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">High expression level of soluble SARS spike protein mediated by adenovirus in HEK293 cells.</title><author><name sortKey="Zhong, Fei" sort="Zhong, Fei" uniqKey="Zhong F" first="Fei" last="Zhong">Fei Zhong</name><affiliation><nlm:affiliation>Department of Basic Veterinary Medicine, School of Animal Science and Technology, Hebei Agricultural University, Baoding 071001, Hebei Province, China. feizhong2000@yahoo.com</nlm:affiliation></affiliation></author><author><name sortKey="Zhong, Zhen Yu" sort="Zhong, Zhen Yu" uniqKey="Zhong Z" first="Zhen-Yu" last="Zhong">Zhen-Yu Zhong</name></author><author><name sortKey="Liang, Shuang" sort="Liang, Shuang" uniqKey="Liang S" first="Shuang" last="Liang">Shuang Liang</name></author><author><name sortKey="Li, Xiu Jin" sort="Li, Xiu Jin" uniqKey="Li X" first="Xiu-Jin" last="Li">Xiu-Jin Li</name></author></analytic><series><title level="j">World journal of gastroenterology</title><idno type="ISSN">1007-9327</idno><imprint><date when="2006" type="published">2006</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Adenoviridae (genetics)</term><term>Blotting, Western</term><term>Cell Line</term><term>Cell Survival</term><term>Gene Expression Regulation, Viral</term><term>Genetic Vectors</term><term>Humans</term><term>Membrane Glycoproteins (biosynthesis)</term><term>Membrane Glycoproteins (genetics)</term><term>Plasmids</term><term>Recombinant Proteins (analysis)</term><term>SARS Virus</term><term>Spike Glycoprotein, Coronavirus</term><term>Viral Envelope Proteins (biosynthesis)</term><term>Viral Envelope Proteins (genetics)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="analysis" xml:lang="en"><term>Recombinant Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="biosynthesis" xml:lang="en"><term>Membrane Glycoproteins</term><term>Viral Envelope Proteins</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Adenoviridae</term><term>Membrane Glycoproteins</term><term>Viral Envelope Proteins</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Blotting, Western</term><term>Cell Line</term><term>Cell Survival</term><term>Gene Expression Regulation, Viral</term><term>Genetic Vectors</term><term>Humans</term><term>Plasmids</term><term>SARS Virus</term><term>Spike Glycoprotein, Coronavirus</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">To develop a highly efficacious method for preparation of soluble SARS S-protein using adenovirus vector to meet the requirement for S-protein investigation.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">16552820</PMID><DateCompleted><Year>2006</Year><Month>08</Month><Day>09</Day></DateCompleted><DateRevised><Year>2020</Year><Month>04</Month><Day>15</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Print">1007-9327</ISSN><JournalIssue CitedMedium="Print"><Volume>12</Volume><Issue>9</Issue><PubDate><Year>2006</Year><Month>Mar</Month><Day>07</Day></PubDate></JournalIssue><Title>World journal of gastroenterology</Title><ISOAbbreviation>World J. Gastroenterol.</ISOAbbreviation></Journal><ArticleTitle>High expression level of soluble SARS spike protein mediated by adenovirus in HEK293 cells.</ArticleTitle><Pagination><MedlinePgn>1452-7</MedlinePgn></Pagination><Abstract><AbstractText Label="AIM" NlmCategory="OBJECTIVE">To develop a highly efficacious method for preparation of soluble SARS S-protein using adenovirus vector to meet the requirement for S-protein investigation.</AbstractText><AbstractText Label="METHODS" NlmCategory="METHODS">The human adenovirus vector was used to express the soluble S-protein (corresponding to 1 approximately 1190 amino acids) fused with Myc/His tag using codon-optimized gene construct in HEK239 cells. The recombinant adenovirus bearing S-protein gene was generated by ligation method. The expressed S-protein with Myc/His tag was purified from culture medium with Ni-NTA agarose beads followed by dialysis. The S-protein was detected by Western blot and its biologic activity was analyzed by binding to Vero cells.</AbstractText><AbstractText Label="RESULTS" NlmCategory="RESULTS">Under the conditions of infection dose (MOI of 50) and expression time (48 h), the high-level expression of S-protein was obtained. The expression level was determined to be approximately 75 microg/10(6) cells after purification. Purified soluble S-protein was readily detected by Western blot with anti-Myc antibody and showed the ability to bind to surface of Vero cells,demonstrating that the soluble S-protein could remain the biologic activity in the native molecule.</AbstractText><AbstractText Label="CONCLUSION" NlmCategory="CONCLUSIONS">The high-level expression of S-protein in HEK293 cells mediated by adenovirus can be achieved under the optimized expression conditions. The proteins possess the biologic activity, which lays a foundation for further investigation of S-protein biological function.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Zhong</LastName><ForeName>Fei</ForeName><Initials>F</Initials><AffiliationInfo><Affiliation>Department of Basic Veterinary Medicine, School of Animal Science and Technology, Hebei Agricultural University, Baoding 071001, Hebei Province, China. feizhong2000@yahoo.com</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Zhong</LastName><ForeName>Zhen-Yu</ForeName><Initials>ZY</Initials></Author><Author ValidYN="Y"><LastName>Liang</LastName><ForeName>Shuang</ForeName><Initials>S</Initials></Author><Author ValidYN="Y"><LastName>Li</LastName><ForeName>Xiu-Jin</ForeName><Initials>XJ</Initials></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType></PublicationTypeList></Article><MedlineJournalInfo><Country>United States</Country><MedlineTA>World J Gastroenterol</MedlineTA><NlmUniqueID>100883448</NlmUniqueID><ISSNLinking>1007-9327</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="C578553">MHV surface projection glycoprotein</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D008562">Membrane Glycoproteins</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D011994">Recombinant Proteins</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D064370">Spike Glycoprotein, Coronavirus</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D014759">Viral Envelope Proteins</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="C578557">spike glycoprotein, SARS-CoV</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D000256" MajorTopicYN="N">Adenoviridae</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D015153" MajorTopicYN="N">Blotting, Western</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D002460" MajorTopicYN="N">Cell Line</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D002470" MajorTopicYN="N">Cell Survival</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D015967" MajorTopicYN="N">Gene Expression Regulation, Viral</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D005822" MajorTopicYN="Y">Genetic Vectors</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D006801" MajorTopicYN="N">Humans</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D008562" MajorTopicYN="N">Membrane Glycoproteins</DescriptorName><QualifierName UI="Q000096" MajorTopicYN="Y">biosynthesis</QualifierName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D010957" MajorTopicYN="N">Plasmids</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D011994" MajorTopicYN="N">Recombinant Proteins</DescriptorName><QualifierName UI="Q000032" MajorTopicYN="N">analysis</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D045473" MajorTopicYN="Y">SARS Virus</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D064370" MajorTopicYN="N">Spike Glycoprotein, Coronavirus</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D014759" MajorTopicYN="N">Viral Envelope Proteins</DescriptorName><QualifierName UI="Q000096" MajorTopicYN="Y">biosynthesis</QualifierName><QualifierName UI="Q000235" 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