Expression of erythroid differentiation factor (EDF) in Chinese hamster ovary cells.
Plasmid DNA containing EDF subunit cDNA and mouse dihydrofolate reductase (DHFR) cDNA was transfected into CHO DHFR- cells by the calcium-phosphate method. DHFR positive transformants secreted recombinant EDF (r-EDF) constitutively in an active form and accumulated it in the conditioned medium. Furthermore, cells which were resistant to methotrexate (MTX : 0.5 microM) secreted r-EDF up to 1 microgram/ml. r-EDF was identical to natural EDF (n-EDF) produced by human acute monocytic leukemia cell line, THP-1, as regards its dimeric structure and a biological activity.
Murata
M
M
Central Research Labortories, Ajinomoto Co., Inc., Kawasaki, Japan.
Onomichi
K
K
Eto
Y
Y
Shibai
H
H
Muramatsu
M
M
D028341:Activins; D000595:Amino Acid Sequence; D000818:Animals; D002454:Cell Differentiation; D002460:Cell Line; D003470:Culture Media; D004247:DNA / Q000235:genetics; D004351:Drug Resistance; D004591:Electrophoresis, Polyacrylamide Gel; D007118:Immunoassay; D007265:Inhibins / Q000096:biosynthesis* / Q000235:genetics; D008727:Methotrexate / Q000494:pharmacology; D010957:Plasmids; D011994:Recombinant Proteins / Q000096:biosynthesis; D013762:Tetrahydrofolate Dehydrogenase / Q000378:metabolism; D014162:Transfection
D003470:Culture Media; D011994:Recombinant Proteins; D028341:Activins; D007265:Inhibins; D004247:DNA; D013762:Tetrahydrofolate Dehydrogenase; D008727:Methotrexate