Production and characterisation of ovine GM-CSF expressed in mammalian and bacterial cells.
A cDNA encoding ovine granulocyte-macrophage colony-stimulating factor (GM-CSF) was isolated and two forms of recombinant ovine GM-CSF were produced. A glycosylated form was produced in mammalian cells infected with a recombinant vaccinia virus encoding ovine GM-CSF. Recombinant ovine GM-CSF was also produced in Escherichia coli and purified by affinity chromatography. Both forms of the protein were detected by ovine GM-CSF-specific monoclonal antibodies, and exhibited activity on ovine bone marrow haemopoetic progenitor cells.
O'Brien
P M
PM
CSIRO Division of Animal Health, Parkville, Australia.
Seow
H F
HF
Entrican
G
G
Coupar
B E
BE
Wood
P R
PR
D000818:Animals; D000911:Antibodies, Monoclonal; D001483:Base Sequence; D015153:Blotting, Western / Q000662:veterinary; D001854:Bone Marrow Cells; D002455:Cell Division; D002460:Cell Line; D002846:Chromatography, Affinity / Q000662:veterinary; D003001:Cloning, Molecular; D003114:Colony-Forming Units Assay / Q000662:veterinary; D017931:DNA Primers / Q000737:chemistry; D018076:DNA, Complementary / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel / Q000662:veterinary; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000096:biosynthesis* / Q000302:isolation & purification / Q000502:physiology; D006801:Humans; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D012756:Sheep / Q000276:immunology*; D013234:Stem Cells / Q000166:cytology; D014162:Transfection; D014616:Vaccinia virus / Q000235:genetics / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D017931:DNA Primers; D018076:DNA, Complementary; D011994:Recombinant Proteins; D016178:Granulocyte-Macrophage Colony-Stimulating Factor