Construction of plasmids expressing Sars-CoV encoding proteins and their effects on transcription of hfgl2 prothrombinase.
Identifieur interne : 001F39 ( Ncbi/Merge ); précédent : 001F38; suivant : 001F40Construction of plasmids expressing Sars-CoV encoding proteins and their effects on transcription of hfgl2 prothrombinase.
Auteurs : Hongwu Wang [République populaire de Chine] ; Meifang Han ; Huaning Yao ; Zhanhui Wang ; Dong Xi ; Weiming Yan ; Jinlin Hou ; Xiaoping Luo ; Qin NingSource :
- Journal of Huazhong University of Science and Technology. Medical sciences = Hua zhong ke ji da xue xue bao. Yi xue Ying De wen ban = Huazhong keji daxue xuebao. Yixue Yingdewen ban [ 1672-0733 ] ; 2009.
Descripteurs français
- KwdFr :
- Animaux, Cellules CHO, Cricetinae, Cricetulus, Fibrinogène (biosynthèse), Fibrinogène (génétique), Glycoprotéine de spicule des coronavirus, Glycoprotéines membranaires (biosynthèse), Glycoprotéines membranaires (génétique), Humains, Plasmides, Protéines de l'enveloppe virale (biosynthèse), Protéines de l'enveloppe virale (génétique), Protéines de la matrice virale (biosynthèse), Protéines de la matrice virale (génétique), Protéines nucléocapside (biosynthèse), Protéines nucléocapside (génétique), Protéines recombinantes (biosynthèse), Syndrome respiratoire aigu sévère (virologie), Thromboplastine (génétique), Thromboplastine (métabolisme), Transcription génétique (génétique), Transfection, Vecteurs génétiques (génétique), Virus du SRAS (génétique), Virus du SRAS (métabolisme).
- MESH :
- biosynthèse : Fibrinogène, Glycoprotéines membranaires, Protéines de l'enveloppe virale, Protéines de la matrice virale, Protéines nucléocapside, Protéines recombinantes.
- génétique : Fibrinogène, Glycoprotéines membranaires, Protéines de l'enveloppe virale, Protéines de la matrice virale, Protéines nucléocapside, Thromboplastine, Transcription génétique, Vecteurs génétiques, Virus du SRAS.
- métabolisme : Thromboplastine, Virus du SRAS.
- virologie : Syndrome respiratoire aigu sévère.
- Animaux, Cellules CHO, Cricetinae, Cricetulus, Glycoprotéine de spicule des coronavirus, Humains, Plasmides, Transfection.
English descriptors
- KwdEn :
- Animals, CHO Cells, Cricetinae, Cricetulus, Fibrinogen (biosynthesis), Fibrinogen (genetics), Genetic Vectors (genetics), Humans, Membrane Glycoproteins (biosynthesis), Membrane Glycoproteins (genetics), Nucleocapsid Proteins (biosynthesis), Nucleocapsid Proteins (genetics), Plasmids, Recombinant Proteins (biosynthesis), SARS Virus (genetics), SARS Virus (metabolism), Severe Acute Respiratory Syndrome (virology), Spike Glycoprotein, Coronavirus, Thromboplastin (genetics), Thromboplastin (metabolism), Transcription, Genetic (genetics), Transfection, Viral Envelope Proteins (biosynthesis), Viral Envelope Proteins (genetics), Viral Matrix Proteins (biosynthesis), Viral Matrix Proteins (genetics).
- MESH :
- chemical , biosynthesis : Fibrinogen, Membrane Glycoproteins, Nucleocapsid Proteins, Recombinant Proteins, Viral Envelope Proteins, Viral Matrix Proteins.
- chemical , genetics : Fibrinogen, Membrane Glycoproteins, Nucleocapsid Proteins, Thromboplastin, Viral Envelope Proteins, Viral Matrix Proteins.
- genetics : Genetic Vectors, SARS Virus, Transcription, Genetic.
- metabolism : SARS Virus, Thromboplastin.
- virology : Severe Acute Respiratory Syndrome.
- Animals, CHO Cells, Cricetinae, Cricetulus, Humans, Plasmids, Spike Glycoprotein, Coronavirus, Transfection.
Abstract
SARS coronavirus (SARS-CoV) is the etiologic agent of severe acute respiratory syndrome. The aim of this study was to construct Sars-CoV membrane (M), nucleocapsid (N) and spike 2 (S2) gene eukaryotic expression plasmids, and identify their expression in vitro. Gene fragments encoding N protein, M protein and S2 protein of SARS-CoV were amplified by PCR using cDNA obtained from lung samples of SARS patients as template, and subcloned into pcDNA3.1 vector to form eukaryotic expression plasmids. SARS-CoV protein eukaryotic expression plasmids were transfected respectively into CHO cells. Immunohistochemistry was employed to detect the expression of the structural proteins of SARS-CoV in transfected cells. SARS-CoV protein eukaryotic expression plasmids were successfully constructed by identification with digestion of restriction enzymes and sequencing. M, N and S2 proteins of SARS-CoV were detected in the cytoplasm of transfected CHO cells. It was concluded that these recombinant eukaryotic expression plasmids were constructed successfully, and SARS-CoV encoding proteins could activate transcription and expression of hfgl2 gene.
DOI: 10.1007/s11596-009-0311-1
PubMed: 19513614
Links toward previous steps (curation, corpus...)
- to stream PubMed, to step Corpus: 001886
- to stream PubMed, to step Curation: 001886
- to stream PubMed, to step Checkpoint: 001919
Links to Exploration step
pubmed:19513614Le document en format XML
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<term>CHO Cells</term>
<term>Cricetinae</term>
<term>Cricetulus</term>
<term>Fibrinogen (biosynthesis)</term>
<term>Fibrinogen (genetics)</term>
<term>Genetic Vectors (genetics)</term>
<term>Humans</term>
<term>Membrane Glycoproteins (biosynthesis)</term>
<term>Membrane Glycoproteins (genetics)</term>
<term>Nucleocapsid Proteins (biosynthesis)</term>
<term>Nucleocapsid Proteins (genetics)</term>
<term>Plasmids</term>
<term>Recombinant Proteins (biosynthesis)</term>
<term>SARS Virus (genetics)</term>
<term>SARS Virus (metabolism)</term>
<term>Severe Acute Respiratory Syndrome (virology)</term>
<term>Spike Glycoprotein, Coronavirus</term>
<term>Thromboplastin (genetics)</term>
<term>Thromboplastin (metabolism)</term>
<term>Transcription, Genetic (genetics)</term>
<term>Transfection</term>
<term>Viral Envelope Proteins (biosynthesis)</term>
<term>Viral Envelope Proteins (genetics)</term>
<term>Viral Matrix Proteins (biosynthesis)</term>
<term>Viral Matrix Proteins (genetics)</term>
</keywords>
<keywords scheme="KwdFr" xml:lang="fr"><term>Animaux</term>
<term>Cellules CHO</term>
<term>Cricetinae</term>
<term>Cricetulus</term>
<term>Fibrinogène (biosynthèse)</term>
<term>Fibrinogène (génétique)</term>
<term>Glycoprotéine de spicule des coronavirus</term>
<term>Glycoprotéines membranaires (biosynthèse)</term>
<term>Glycoprotéines membranaires (génétique)</term>
<term>Humains</term>
<term>Plasmides</term>
<term>Protéines de l'enveloppe virale (biosynthèse)</term>
<term>Protéines de l'enveloppe virale (génétique)</term>
<term>Protéines de la matrice virale (biosynthèse)</term>
<term>Protéines de la matrice virale (génétique)</term>
<term>Protéines nucléocapside (biosynthèse)</term>
<term>Protéines nucléocapside (génétique)</term>
<term>Protéines recombinantes (biosynthèse)</term>
<term>Syndrome respiratoire aigu sévère (virologie)</term>
<term>Thromboplastine (génétique)</term>
<term>Thromboplastine (métabolisme)</term>
<term>Transcription génétique (génétique)</term>
<term>Transfection</term>
<term>Vecteurs génétiques (génétique)</term>
<term>Virus du SRAS (génétique)</term>
<term>Virus du SRAS (métabolisme)</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="biosynthesis" xml:lang="en"><term>Fibrinogen</term>
<term>Membrane Glycoproteins</term>
<term>Nucleocapsid Proteins</term>
<term>Recombinant Proteins</term>
<term>Viral Envelope Proteins</term>
<term>Viral Matrix Proteins</term>
</keywords>
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<term>Membrane Glycoproteins</term>
<term>Nucleocapsid Proteins</term>
<term>Thromboplastin</term>
<term>Viral Envelope Proteins</term>
<term>Viral Matrix Proteins</term>
</keywords>
<keywords scheme="MESH" qualifier="biosynthèse" xml:lang="fr"><term>Fibrinogène</term>
<term>Glycoprotéines membranaires</term>
<term>Protéines de l'enveloppe virale</term>
<term>Protéines de la matrice virale</term>
<term>Protéines nucléocapside</term>
<term>Protéines recombinantes</term>
</keywords>
<keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Genetic Vectors</term>
<term>SARS Virus</term>
<term>Transcription, Genetic</term>
</keywords>
<keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Fibrinogène</term>
<term>Glycoprotéines membranaires</term>
<term>Protéines de l'enveloppe virale</term>
<term>Protéines de la matrice virale</term>
<term>Protéines nucléocapside</term>
<term>Thromboplastine</term>
<term>Transcription génétique</term>
<term>Vecteurs génétiques</term>
<term>Virus du SRAS</term>
</keywords>
<keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>SARS Virus</term>
<term>Thromboplastin</term>
</keywords>
<keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Thromboplastine</term>
<term>Virus du SRAS</term>
</keywords>
<keywords scheme="MESH" qualifier="virologie" xml:lang="fr"><term>Syndrome respiratoire aigu sévère</term>
</keywords>
<keywords scheme="MESH" qualifier="virology" xml:lang="en"><term>Severe Acute Respiratory Syndrome</term>
</keywords>
<keywords scheme="MESH" xml:lang="en"><term>Animals</term>
<term>CHO Cells</term>
<term>Cricetinae</term>
<term>Cricetulus</term>
<term>Humans</term>
<term>Plasmids</term>
<term>Spike Glycoprotein, Coronavirus</term>
<term>Transfection</term>
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<keywords scheme="MESH" xml:lang="fr"><term>Animaux</term>
<term>Cellules CHO</term>
<term>Cricetinae</term>
<term>Cricetulus</term>
<term>Glycoprotéine de spicule des coronavirus</term>
<term>Humains</term>
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<front><div type="abstract" xml:lang="en">SARS coronavirus (SARS-CoV) is the etiologic agent of severe acute respiratory syndrome. The aim of this study was to construct Sars-CoV membrane (M), nucleocapsid (N) and spike 2 (S2) gene eukaryotic expression plasmids, and identify their expression in vitro. Gene fragments encoding N protein, M protein and S2 protein of SARS-CoV were amplified by PCR using cDNA obtained from lung samples of SARS patients as template, and subcloned into pcDNA3.1 vector to form eukaryotic expression plasmids. SARS-CoV protein eukaryotic expression plasmids were transfected respectively into CHO cells. Immunohistochemistry was employed to detect the expression of the structural proteins of SARS-CoV in transfected cells. SARS-CoV protein eukaryotic expression plasmids were successfully constructed by identification with digestion of restriction enzymes and sequencing. M, N and S2 proteins of SARS-CoV were detected in the cytoplasm of transfected CHO cells. It was concluded that these recombinant eukaryotic expression plasmids were constructed successfully, and SARS-CoV encoding proteins could activate transcription and expression of hfgl2 gene.</div>
</front>
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<DateCompleted><Year>2010</Year>
<Month>10</Month>
<Day>21</Day>
</DateCompleted>
<DateRevised><Year>2020</Year>
<Month>04</Month>
<Day>15</Day>
</DateRevised>
<Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Print">1672-0733</ISSN>
<JournalIssue CitedMedium="Print"><Volume>29</Volume>
<Issue>3</Issue>
<PubDate><Year>2009</Year>
<Month>Jun</Month>
</PubDate>
</JournalIssue>
<Title>Journal of Huazhong University of Science and Technology. Medical sciences = Hua zhong ke ji da xue xue bao. Yi xue Ying De wen ban = Huazhong keji daxue xuebao. Yixue Yingdewen ban</Title>
<ISOAbbreviation>J. Huazhong Univ. Sci. Technol. Med. Sci.</ISOAbbreviation>
</Journal>
<ArticleTitle>Construction of plasmids expressing Sars-CoV encoding proteins and their effects on transcription of hfgl2 prothrombinase.</ArticleTitle>
<Pagination><MedlinePgn>318-23</MedlinePgn>
</Pagination>
<ELocationID EIdType="doi" ValidYN="Y">10.1007/s11596-009-0311-1</ELocationID>
<Abstract><AbstractText>SARS coronavirus (SARS-CoV) is the etiologic agent of severe acute respiratory syndrome. The aim of this study was to construct Sars-CoV membrane (M), nucleocapsid (N) and spike 2 (S2) gene eukaryotic expression plasmids, and identify their expression in vitro. Gene fragments encoding N protein, M protein and S2 protein of SARS-CoV were amplified by PCR using cDNA obtained from lung samples of SARS patients as template, and subcloned into pcDNA3.1 vector to form eukaryotic expression plasmids. SARS-CoV protein eukaryotic expression plasmids were transfected respectively into CHO cells. Immunohistochemistry was employed to detect the expression of the structural proteins of SARS-CoV in transfected cells. SARS-CoV protein eukaryotic expression plasmids were successfully constructed by identification with digestion of restriction enzymes and sequencing. M, N and S2 proteins of SARS-CoV were detected in the cytoplasm of transfected CHO cells. It was concluded that these recombinant eukaryotic expression plasmids were constructed successfully, and SARS-CoV encoding proteins could activate transcription and expression of hfgl2 gene.</AbstractText>
</Abstract>
<AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Wang</LastName>
<ForeName>Hongwu</ForeName>
<Initials>H</Initials>
<AffiliationInfo><Affiliation>Department of Infectious Disease, Huazhong University of Science and Technology, Wuhan, 430030, China. hongwuwang@126.com</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Han</LastName>
<ForeName>Meifang</ForeName>
<Initials>M</Initials>
</Author>
<Author ValidYN="Y"><LastName>Yao</LastName>
<ForeName>Huaning</ForeName>
<Initials>H</Initials>
</Author>
<Author ValidYN="Y"><LastName>Wang</LastName>
<ForeName>Zhanhui</ForeName>
<Initials>Z</Initials>
</Author>
<Author ValidYN="Y"><LastName>Xi</LastName>
<ForeName>Dong</ForeName>
<Initials>D</Initials>
</Author>
<Author ValidYN="Y"><LastName>Yan</LastName>
<ForeName>Weiming</ForeName>
<Initials>W</Initials>
</Author>
<Author ValidYN="Y"><LastName>Hou</LastName>
<ForeName>Jinlin</ForeName>
<Initials>J</Initials>
</Author>
<Author ValidYN="Y"><LastName>Luo</LastName>
<ForeName>Xiaoping</ForeName>
<Initials>X</Initials>
</Author>
<Author ValidYN="Y"><LastName>Ning</LastName>
<ForeName>Qin</ForeName>
<Initials>Q</Initials>
</Author>
</AuthorList>
<Language>eng</Language>
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<PublicationType UI="D013485">Research Support, Non-U.S. Gov't</PublicationType>
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<ArticleDate DateType="Electronic"><Year>2009</Year>
<Month>06</Month>
<Day>10</Day>
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<MedlineJournalInfo><Country>China</Country>
<MedlineTA>J Huazhong Univ Sci Technolog Med Sci</MedlineTA>
<NlmUniqueID>101169627</NlmUniqueID>
<ISSNLinking>1672-0733</ISSNLinking>
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<ChemicalList><Chemical><RegistryNumber>0</RegistryNumber>
<NameOfSubstance UI="C095202">FGL2 protein, human</NameOfSubstance>
</Chemical>
<Chemical><RegistryNumber>0</RegistryNumber>
<NameOfSubstance UI="D008562">Membrane Glycoproteins</NameOfSubstance>
</Chemical>
<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="D064370">Spike Glycoprotein, Coronavirus</NameOfSubstance>
</Chemical>
<Chemical><RegistryNumber>0</RegistryNumber>
<NameOfSubstance UI="D014759">Viral Envelope Proteins</NameOfSubstance>
</Chemical>
<Chemical><RegistryNumber>0</RegistryNumber>
<NameOfSubstance UI="D014763">Viral Matrix Proteins</NameOfSubstance>
</Chemical>
<Chemical><RegistryNumber>0</RegistryNumber>
<NameOfSubstance UI="C099602">nucleocapsid protein, Coronavirus</NameOfSubstance>
</Chemical>
<Chemical><RegistryNumber>0</RegistryNumber>
<NameOfSubstance UI="C578557">spike glycoprotein, SARS-CoV</NameOfSubstance>
</Chemical>
<Chemical><RegistryNumber>108502-71-2</RegistryNumber>
<NameOfSubstance UI="C067997">M protein, Coronavirus</NameOfSubstance>
</Chemical>
<Chemical><RegistryNumber>9001-32-5</RegistryNumber>
<NameOfSubstance UI="D005340">Fibrinogen</NameOfSubstance>
</Chemical>
<Chemical><RegistryNumber>9035-58-9</RegistryNumber>
<NameOfSubstance UI="D013925">Thromboplastin</NameOfSubstance>
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</MeshHeading>
<MeshHeading><DescriptorName UI="D016466" MajorTopicYN="N">CHO Cells</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D006224" MajorTopicYN="N">Cricetinae</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D003412" MajorTopicYN="N">Cricetulus</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D005340" MajorTopicYN="N">Fibrinogen</DescriptorName>
<QualifierName UI="Q000096" MajorTopicYN="N">biosynthesis</QualifierName>
<QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D005822" MajorTopicYN="N">Genetic Vectors</DescriptorName>
<QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D006801" MajorTopicYN="N">Humans</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D008562" MajorTopicYN="N">Membrane Glycoproteins</DescriptorName>
<QualifierName UI="Q000096" MajorTopicYN="N">biosynthesis</QualifierName>
<QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName>
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<MeshHeading><DescriptorName UI="D019590" MajorTopicYN="N">Nucleocapsid Proteins</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="Q000096" MajorTopicYN="N">biosynthesis</QualifierName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D045473" MajorTopicYN="N">SARS Virus</DescriptorName>
<QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName>
<QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D045169" MajorTopicYN="N">Severe Acute Respiratory Syndrome</DescriptorName>
<QualifierName UI="Q000821" MajorTopicYN="N">virology</QualifierName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D064370" MajorTopicYN="N">Spike Glycoprotein, Coronavirus</DescriptorName>
</MeshHeading>
<MeshHeading><DescriptorName UI="D013925" MajorTopicYN="N">Thromboplastin</DescriptorName>
<QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName>
<QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName>
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