Abstract
The gene encoding the major nonstructural (NS-1) protein of minute virus of mice (MVM) has been expressed in insect cells using a baculovirus expression system. This 83-kDa polypeptide was found to be localized in the soluble (cytosolic) fraction in insect cells, in contrast with the nuclear localization of NS-1 expressed in MVM-infected mouse LA-9 cells. The protein was purified by immunoaffinity chromatography using a monoclonal antibody (MAb) prepared to an NS-1 fusion peptide [(Yeung et al., Virology 185, 35-45 (1991)]. Recombinant NS-1 was eluted using either low pH or a synthetic peptide corresponding to the epitope of the MAb. The peptide-eluted material is greater than 95% pure and biologically active in that it has ATPase activity and ATP-dependent helicase activity as determined by a strand displacement assay.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Adenosine Triphosphatases / genetics*
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Adenosine Triphosphatases / isolation & purification
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Adenosine Triphosphatases / metabolism
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Adenosine Triphosphate / metabolism
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Amino Acid Sequence
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Animals
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Antibodies, Monoclonal
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Baculoviridae / genetics
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Capsid / genetics*
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Capsid / isolation & purification
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Capsid / metabolism
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Cell Line
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Chromatography, Affinity
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Cloning, Molecular
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Deoxyadenine Nucleotides / metabolism
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Electrophoresis, Polyacrylamide Gel
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Genes, Viral
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Insecta
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Minute Virus of Mice / enzymology
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Minute Virus of Mice / genetics*
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Molecular Sequence Data
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Molecular Weight
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Multienzyme Complexes / genetics
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Multienzyme Complexes / isolation & purification
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Multienzyme Complexes / metabolism
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Peptides / chemical synthesis
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Peptides / immunology
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Plasmids
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Recombinant Proteins / isolation & purification
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Recombinant Proteins / metabolism
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Substrate Specificity
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Transfection*
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Viral Core Proteins / genetics*
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Viral Core Proteins / isolation & purification
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Viral Core Proteins / metabolism
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Viral Nonstructural Proteins
Substances
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Antibodies, Monoclonal
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Deoxyadenine Nucleotides
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Multienzyme Complexes
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Peptides
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Recombinant Proteins
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Viral Core Proteins
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Viral Nonstructural Proteins
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Adenosine Triphosphate
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Adenosine Triphosphatases
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2'-deoxyadenosine triphosphate