Recombinant murine GM-CSF from E. coli has biological activity and is neutralized by a specific antiserum.
We report the production and characterization of a mouse granulocyte-macrophage colony stimulating factor (mGM-CSF) made in Escherichia coli. The synthesis of mGM-CSF was directed by a plasmid containing a gene isolated from the EL-4 cell line. After induction of expression and accumulation of the protein in E. coli, mGM-CSF accounted for 10% of total cellular protein. This recombinant mGM-CSF was purified to 90% homogeneity by chaotrope extraction and gel filtration. Recombinant mGM-CSF, like the native molecule, stimulates the growth of granulocyte and macrophage colonies in serum-free cultures of mouse bone marrow cells. Antibodies raised against recombinant mGM-CSF not only reacted with the recombinant protein but also neutralized the biological activity of both native and recombinant mGM-CSF. These results indicate that the functional structure of the recombinant protein is similar to that of native mGM-CSF.
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001681:Biological Assay; D003115:Colony-Stimulating Factors / Q000235:genetics* / Q000276:immunology / Q000502:physiology; D004926:Escherichia coli; D005786:Gene Expression Regulation; D005796:Genes; D006098:Granulocytes; D008264:Macrophages; D051379:Mice; D008970:Molecular Weight; D009500:Neutralization Tests; D011994:Recombinant Proteins / Q000276:immunology / Q000502:physiology*
D003115:Colony-Stimulating Factors; D011994:Recombinant Proteins