Molecular cloning of a cDNA encoding a human macrophage migration inhibitory factor

Proc Natl Acad Sci U S A. 1989 Oct;86(19):7522-6. doi: 10.1073/pnas.86.19.7522.

Abstract

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.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Cell Line
  • Cloning, Molecular*
  • DNA / genetics*
  • DNA Transposable Elements
  • Humans
  • Macrophage Migration-Inhibitory Factors / genetics*
  • Molecular Sequence Data
  • Mutation
  • Transfection

Substances

  • DNA Transposable Elements
  • Macrophage Migration-Inhibitory Factors
  • DNA

Associated data

  • GENBANK/M25639