Development of a vector system for the expression of bioengineered proteins

Clin Chem. 1989 Jul;35(7 Suppl):B7-12.

Abstract

The low natural abundance of many proteins is a major factor in preventing their development as therapeutic or diagnostic tools. To circumvent this barrier, we have used synthetic oligonucleotide technology to construct a gene based on the sequence of a cDNA for human interleukin 6 (IL-6). The synthetic gene encodes a cysteine-free, bioengineered rIL-6 protein, which is expressed at high concentrations in Escherichia coli as a tripartite fusion protein. Cleavage of the fusion protein with collagenase (EC 3.4.24.8) releases a 23-kDa rIL-6 protein that can be easily purified to homogeneity. This rIL-6 protein displays a range of biological activities similar to those of natural human IL-6, as demonstrated by its ability to (a) protect cells from viral infection and (b) stimulate the synthesis of fibrinogen in rat FAZA cells.

Publication types

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

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Escherichia coli / genetics
  • Genetic Vectors*
  • Interleukin-6
  • Interleukins / biosynthesis*
  • Interleukins / genetics
  • Interleukins / isolation & purification
  • Interleukins / pharmacology
  • Liver / metabolism
  • Molecular Sequence Data
  • Plasmids
  • Rats
  • Recombinant Fusion Proteins / biosynthesis
  • Recombinant Fusion Proteins / isolation & purification
  • Recombinant Fusion Proteins / pharmacology
  • Viral Plaque Assay

Substances

  • Interleukin-6
  • Interleukins
  • Recombinant Fusion Proteins