title: High-level expression in Escherichia coli of a soluble and fully active recombinant interleukin-1 beta.
abstract: A complementary DNA sequence encoding monocyte interleukin-1 (IL-1), beta form/pI7, was expressed in Escherichia coli. Recombinant plasmid pDP516 was constructed by cloning and rebuilding the mature IL-1 coding sequence into an E. coli expression vector. Bacteria transformed with pDP516 constitutively produced recombinant IL-1 (r-IL-1) at 15-20% of total E. coli protein. The r-IL-1 was found to be in the soluble fraction of sonicated E. coli Bacterial r-IL-1 (DP516) has been purified to homogeneity by anion exchange and sizing column chromatography, with an apparent molecular weight of 17,500. The identity of the purified r-IL-1 was confirmed by amino acid and DNA sequencing analyses. Purified recombinant IL-1 DP516 exhibits biological activity similar to that of native monocyte IL-1 (3 approximately 4 X 10(7) units/mg). An amino-terminal deletion mutant completely abolishes the biological activity, indicating that the integrity of the IL-1 molecule might be important for its function.
authors.forename: J J
authors.initials: JJ
authors.lastname: Huang
authors.forename: R C
authors.initials: RC
authors.lastname: Newton
authors.forename: K
authors.initials: K
authors.lastname: Pezzella
authors.forename: M
authors.initials: M
authors.lastname: Covington
authors.forename: T
authors.initials: T
authors.lastname: Tamblyn
authors.forename: S J
authors.initials: SJ
authors.lastname: Rutlege
authors.forename: J
authors.initials: J
authors.lastname: Gray
authors.forename: M
authors.initials: M
authors.lastname: Kelley
authors.forename: Y
authors.initials: Y
authors.lastname: Lin
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D001681:Biological Assay; D003001:Cloning, Molecular / Q000379:methods; D004926:Escherichia coli / Q000235:genetics; D005786:Gene Expression Regulation; D006801:Humans; D007375:Interleukin-1 / Q000235:genetics* / Q000302:isolation & purification; D010957:Plasmids; D011994:Recombinant Proteins; D012995:Solubility
chemical_list: D007375:Interleukin-1; D011994:Recombinant Proteins