Abstract
A new transfection system for influenza virus was developed using the clone 76 cell line, in which the viral RNA polymerase and nucleoprotein (NP) genes can be expressed in response to dexamethasone. Ribonucleoprotein (RNP) complexes were reconstituted by expressing proteins from a chimeric NS-chloramphenicol acetyltransferase (CAT) RNA consisting of the full-length negative-strand RNA of the CAT gene positioned between the 5'- and 3'-terminal sequences of influenza virus RNA segment 8, and purifying NP from an NP gene-expressing Escherichia coli strain. When the reconstituted RNP was transfected into clone 76 cells, CAT was produced only when the synthesis of the three RNA polymerase subunits and NP was induced by treatment with dexamethasone.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Base Sequence
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Cell Line
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Chloramphenicol O-Acetyltransferase / genetics
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Cloning, Molecular
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DNA, Viral
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DNA-Directed RNA Polymerases / genetics*
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DNA-Directed RNA Polymerases / metabolism
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Fluorescent Antibody Technique
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Gene Expression
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Influenza A virus / enzymology
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Influenza A virus / genetics*
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Mice
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Molecular Sequence Data
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Nucleocapsid Proteins
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Nucleoproteins / genetics*
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Nucleoproteins / metabolism
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RNA, Viral / metabolism
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RNA-Binding Proteins*
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Transcription, Genetic*
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Transfection
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Viral Core Proteins / genetics*
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Viral Core Proteins / metabolism
Substances
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DNA, Viral
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NP protein, Influenza A virus
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Nucleocapsid Proteins
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Nucleoproteins
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RNA, Viral
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RNA-Binding Proteins
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Viral Core Proteins
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Chloramphenicol O-Acetyltransferase
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DNA-Directed RNA Polymerases