Genomic cloning and heterologous expression of human differentiation-stimulating factor.
Differentiation-stimulating factor (D-factor) purified from mouse Ehrlich ascites cells was sequenced partially and found to be almost identical to leukemia inhibitory factor (LIF) from mouse Krebs II ascites cells. In comparison to LIF, D-factor had an additional amino-terminal serine residue. Using synthetic oligonucleotide probes designed from the murine D-factor sequence, we cloned the human gene encoding D-factor. A partial D-factor cDNA was cloned from COS-1 cells transfected with the human D-factor gene under the control of a heterologous promoter. We used this cDNA to construct a vector for direct expression of the protein in Escherichia coli. A mammalian cell expression vector was constructed using the signal sequence of interferon-alpha A linked to the D-factor cDNA. Both forms of recombinant human D-factor were active on the murine myeloid leukemia cell line M1 in a dose- and time-dependent manner for the inhibition of [3H]thymidine incorporation, and also induced phagocytosis, Fc receptor expression, and prostaglandin E2 synthesis by M1 cells.
Lowe
D G
DG
Department of Molecular Biology, Genentech, Inc., South San Francisco, CA 94080.
Nunes
W
W
Bombara
M
M
McCabe
S
S
Ranges
G E
GE
Henzel
W
W
Tomida
M
M
Yamamoto-Yamaguchi
Y
Y
Hozumi
M
M
Goeddel
D V
DV
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D005091:Exons; D006131:Growth Inhibitors / Q000235:genetics*; D006801:Humans; D015850:Interleukin-6; D053605:Leukemia Inhibitory Factor; D008222:Lymphokines / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D051379:Mice; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D012313:RNA / Q000096:biosynthesis / Q000235:genetics; D012689:Sequence Homology, Nucleic Acid; D014162:Transfection
D006131:Growth Inhibitors; D015850:Interleukin-6; C507837:LIF protein, human; D053605:Leukemia Inhibitory Factor; C507838:Lif protein, mouse; D008222:Lymphokines; D015345:Oligonucleotide Probes; D012313:RNA; D004247:DNA