Abstract
The expression and partial purification of recombinant 12 kDa B cell growth factor are reported. The polypeptide was derived from the genomic sequence of the gene (BCGF 1) which is here shown to be a single copy gene that localizes to human chromosome 16. When expressed as a glutathione S-transferase fusion protein in E. coli, the protein appears as a 38 kDa polypeptide in Western blot analysis using a peptide antibody. The purified fusion protein stimulates the proliferation of activated human B cells in a dose-dependent manner, and the active site resides within the 104 carboxy-terminal amino acids. The availability of biologically active recombinant 12 kDa B cell growth factor will enable its evaluation in B cell growth regulation, and provides a new means of in vitro culturing of human B lymphocytes.
Publication types
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Comparative Study
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Animals
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B-Lymphocytes / drug effects
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B-Lymphocytes / immunology*
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Base Sequence
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Binding Sites
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Blotting, Southern
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Blotting, Western
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Cells, Cultured
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Chromosome Mapping
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Chromosomes, Human, Pair 16*
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DNA / blood
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DNA / isolation & purification
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DNA Primers
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Gene Expression*
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Glutathione Transferase / biosynthesis
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Humans
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Hybrid Cells
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Interleukin-4 / biosynthesis*
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Interleukin-4 / genetics
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Interleukin-4 / pharmacology*
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Leukocytes / metabolism*
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Lymphocyte Activation*
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Molecular Sequence Data
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Molecular Weight
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Peptides / chemical synthesis
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Peptides / chemistry
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Peptides / immunology
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Polymerase Chain Reaction
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Recombinant Fusion Proteins / biosynthesis
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Recombinant Proteins / pharmacology
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Restriction Mapping
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Rodentia
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Sequence Homology, Amino Acid
Substances
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DNA Primers
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Peptides
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Recombinant Fusion Proteins
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Recombinant Proteins
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Interleukin-4
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DNA
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Glutathione Transferase