Molecular cloning of a cDNA encoding a human macrophage migration inhibitory factor.
A cDNA encoding a human macrophage migration inhibitory factor (MIF) was isolated, through functional expression cloning in COS-1 cells, from a cDNA library prepared from a lectin-stimulated T-cell hybridoma, T-CEMB. The 115-amino acid polypeptide encoded by the MIF cDNA (p7-1) was effectively released from the transfected COS-1 cells and yielded readily detectable MIF activity in the culture supernatant despite the apparent lack of a classical protein secretory sequence. Insertional mutational analysis and elution of MIF activity from polyacrylamide gel slices demonstrated that the Mr 12,000 protein with MIF activity released by the COS-1 cells is encoded by p7-1. The p7-1 cDNA hybridized with a 700-base mRNA expressed by Con-A-stimulated lymphocytes but not unstimulated lymphocytes. The availability of the MIF cDNA clone and recombinant MIF will facilitate the analysis of the role of this lymphokine in cell-mediated immunity, immunoregulation, and inflammation.
Weiser
W Y
WY
Department of Medicine, Harvard Medical School, Boston, MA 02115.
Temple
P A
PA
Witek-Giannotti
J S
JS
Remold
H G
HG
Clark
S C
SC
David
J R
JR
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics*; D004251:DNA Transposable Elements; D006801:Humans; D008263:Macrophage Migration-Inhibitory Factors / Q000235:genetics*; D008969:Molecular Sequence Data; D009154:Mutation; D014162:Transfection
D004251:DNA Transposable Elements; D008263:Macrophage Migration-Inhibitory Factors; D004247:DNA