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Cloning and expression of an interleukin‐1β precursor and its conversion to interleukin‐1β
Author(s) -
Dalbøge H.,
Bayne S.,
Christensen T.,
Hejnæs K.R.
Publication year - 1989
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(89)80259-5
Subject(s) - recombinant dna , cloning (programming) , interleukin , biological activity , monocyte , in vitro , microbiology and biotechnology , interleukin 2 , chemistry , interleukin 4 , molecular cloning , biochemistry , residue (chemistry) , biology , gene expression , gene , cytokine , immunology , programming language , computer science
A gene coding for a N‐terminal precursor of interleukin‐1β (IL‐1β) was cloned and expressed in E. coli. The isolated Met‐Glu‐Ala‐Glu‐IL‐1β precursor was enzymatically converted to IL‐1β by means of dipeptidylaminopeptidase (DAP I). This method ensured a correct N‐terminal residue and the often observed expression of Met‐IL‐1β was thus avoided. The pure and physically homogeneous product exhibited the characteristic properties of natural IL‐1β. The in vitro biological activity was measured in the lymphocyte‐activating factor assay and was compared to that of natural IL‐1β isolated from stimulated monocyte culture using exactly the same purification procedure. The specific biological activity of both products was 2 × 10 −8 U/mg indicating that the recombinant product exhibits full biological activity.