High-level expression in Escherichia coli of a soluble and fully active recombinant interleukin-1 beta.

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.