HIGH-LEVEL EXPRESSION IN ESCHERICHIA-COLI OF A SOLUBLE AND FULLY ACTIVE RECOMBINANT INTERLEUKIN-1-BETA

  • 1 June 1987
    • journal article
    • research article
    • Vol. 4  (3) , 169-181
Abstract
A complementary DNA sequence encoding monocyte interleukin-1 (IL-1), .beta. form/p17, 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 .apprx. 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 analysis. Purified recombinant IL-1 DP516 exhibits biological activity similar to that of native monocyte IL-1 (3 .times. 4 .times. 107 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.

This publication has 0 references indexed in Scilit: