Construction of plasmids expressing Sars-CoV encoding proteins and their effects on transcription of hfgl2 prothrombinase.
Identifieur interne : 001F39 ( Ncbi/Curation ); 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
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pubmed:19513614Le document en format XML
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Medical sciences = Hua zhong ke ji da xue xue bao. Yi xue Ying De wen ban = Huazhong keji daxue xuebao. Yixue Yingdewen ban</title><idno type="ISSN">1672-0733</idno><imprint><date when="2009" type="published">2009</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term><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><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Fibrinogen</term><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></keywords><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><term>Plasmides</term><term>Transfection</term></keywords></textClass></profileDesc></teiHeader><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></TEI></record>Pour manipuler ce document sous Unix (Dilib)
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