Abstract
hM-CSF was reported to have biological activity only in a dimeric form. Using oligonucleotide-directed mutagenesis of hM-CSF (1-149aa) cDNA, we have substituted Ser31 for Cys31 which forms intermolecular disulfide bond in native hM-CSF. The mutant hM-CSF cDNA was expressed in insect BmN cells using baculovirus as a vector under the control of polyhedrin promoter. Biological activity analysis and radioligand receptor assay both showed that there was little difference between the mutant hM-CSF and the native dimeric hM-CSF. These results strongly support that the biologically active human M-CSF in its monomeric form can be expressed in recombinant baculovirus infected insect cells.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Baculoviridae / genetics*
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Base Sequence
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Binding, Competitive
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Cells, Cultured
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DNA, Complementary / genetics
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Genetic Vectors
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Humans
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Insecta
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Macrophages / metabolism
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Mice
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Molecular Sequence Data
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Mutagenesis, Site-Directed
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Occlusion Body Matrix Proteins
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Point Mutation
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Promoter Regions, Genetic / genetics
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Receptor, Macrophage Colony-Stimulating Factor / biosynthesis
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Receptor, Macrophage Colony-Stimulating Factor / genetics*
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Recombinant Proteins / biosynthesis
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Recombinant Proteins / pharmacology*
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Viral Proteins / genetics
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Viral Structural Proteins
Substances
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DNA, Complementary
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Occlusion Body Matrix Proteins
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Recombinant Proteins
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Viral Proteins
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Viral Structural Proteins
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polyhedrin protein, Nucleopolyhedrovirus
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Receptor, Macrophage Colony-Stimulating Factor