Cloning and expression of the cDNA for bovine granulocyte-macrophage colony-stimulating factor.
A sequence encoding bovine granulocyte-macrophage colony-stimulating factor (GM-CSF) has been identified from a concanavalin A-stimulated bovine lymphocyte cDNA library. This sequence was isolated by hybridization with synthetic oligonucleotide probes based upon the human GM-CSF sequence. This bovine cDNA was engineered for expression and secretion of activity into the periplasmic space of E. coli. Periplasmic extracts contain a 14,500-dalton protein and stimulate colony formation of bovine bone marrow progenitor cells. The predicted protein is 70% homologous with human GM-CSF and 55% homologous with murine GM-CSF. Numerous structural features are conserved among these three proteins, such as location of cysteine residues, glycosylation sites, and overall change. The biological activity of bovine GM-CSF is species specific, since recombinant preparations do not cause proliferation of human or murine bone marrow cells. Similarly, murine GM-CSF does not exhibit activity on cells of bovine or human origin. However, human GM-CSF does stimulate colony formation of bovine bone marrow cells, although the specific activity appears reduced when compared to assays on human cells.
Leong
S R
SR
Department of Developmental Biology, South San Francisco, CA 94080.
Flaggs
G M
GM
Lawman
M J
MJ
Gray
P W
PW
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002417:Cattle / Q000235:genetics* / Q000276:immunology; D003001:Cloning, Molecular; D003114:Colony-Forming Units Assay; D003115:Colony-Stimulating Factors / Q000235:genetics* / Q000494:pharmacology; D004247:DNA; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D015723:Gene Library; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D006133:Growth Substances / Q000235:genetics* / Q000494:pharmacology; D015151:Immunoblotting; D051379:Mice; D008969:Molecular Sequence Data; D010957:Plasmids; D012689:Sequence Homology, Nucleic Acid; D013045:Species Specificity
D003115:Colony-Stimulating Factors; D006133:Growth Substances; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D004247:DNA