Open AccessMolecular cloning of a cDNA encoding a human macrophage migration inhibitory factor.
Author(s) -
Weishui Y. Weiser,
Patricia A. Temple,
J Witek-Giannotti,
Heinz G. Remold,
Steven C. Clark,
John R. David
Publication year - 1989
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.86.19.7522
Subject(s) - complementary dna , macrophage migration inhibitory factor , microbiology and biotechnology , biology , cdna library , lymphokine , recombinant dna , molecular cloning , rapid amplification of cdna ends , cloning (programming) , transfection , peptide sequence , biochemistry , gene , cytokine , in vitro , immunology , computer science , programming language
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.