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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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CHO Cells
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Coronavirus M Proteins
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Coronavirus Nucleocapsid Proteins
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Cricetinae
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Cricetulus
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Fibrinogen / biosynthesis
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Fibrinogen / genetics*
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Genetic Vectors / genetics
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Humans
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Membrane Glycoproteins / biosynthesis
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Membrane Glycoproteins / genetics
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Nucleocapsid Proteins / biosynthesis*
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Nucleocapsid Proteins / genetics
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Plasmids
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Recombinant Proteins / biosynthesis
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Severe Acute Respiratory Syndrome / virology
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Severe acute respiratory syndrome-related coronavirus / genetics*
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Severe acute respiratory syndrome-related coronavirus / metabolism
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Spike Glycoprotein, Coronavirus
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Thromboplastin / genetics*
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Thromboplastin / metabolism
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Transcription, Genetic / genetics
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Transfection
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Viral Envelope Proteins / biosynthesis
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Viral Envelope Proteins / genetics
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Viral Matrix Proteins / biosynthesis
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Viral Matrix Proteins / genetics
Substances
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Coronavirus M Proteins
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Coronavirus Nucleocapsid Proteins
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FGL2 protein, human
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M protein, SARS-CoV
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Membrane Glycoproteins
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Nucleocapsid Proteins
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Recombinant Proteins
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Spike Glycoprotein, Coronavirus
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Viral Envelope Proteins
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Viral Matrix Proteins
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spike glycoprotein, SARS-CoV
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Fibrinogen
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Thromboplastin