Cloning of granulocyte colony-stimulating factor cDNA from human macrophages and its expression in Escherichia coli.
Human granulocyte colony-stimulating factor (hG-CSF) cDNA was cloned, by using a synthetic oligonucleotide probe, from an Okayama-Berg cDNA library of lipopolysaccharide-stimulated human peripheral blood macrophages. The cDNA encodes a polypeptide with an amino acid sequence which completely matches that of the known polypeptide with hG-CSF activity derived from human tumor cell lines. Expression in E. coli of high levels of the protein (about 10% of total cellular proteins) was accomplished under control of the trp promoter, and the purified protein was proved to have hG-CSF activity. Our data provide evidence that human peripheral blood macrophages do produce hG-CSF mRNA when stimulated exogenously, suggesting they are the producer of naturally occurring hG-CSF.
D001483:Base Sequence; D003001:Cloning, Molecular; D003115:Colony-Stimulating Factors / Q000096:biosynthesis / Q000235:genetics*; D004247:DNA / Q000032:analysis*; D004926:Escherichia coli / Q000378:metabolism; D016179:Granulocyte Colony-Stimulating Factor; D006098:Granulocytes / Q000502:physiology*; D006801:Humans; D008264:Macrophages / Q000378:metabolism*; D010957:Plasmids
D003115:Colony-Stimulating Factors; D016179:Granulocyte Colony-Stimulating Factor; D004247:DNA