High-level expression of human insulin-like growth factor II in Escherichia coli

J Biotechnol. 1990 Mar;13(4):293-304. doi: 10.1016/0168-1656(90)90077-o.

Abstract

A gene encoding mature human insulin-like growth factor II (IGF-II) was constructed from the modified IGF-II cDNA sequence and two double-stranded synthetic oligodeoxynucleotide linkers. It was fused to a truncated lacZ gene such that IGF-II was expressed as part of C-terminus of beta-galactosidase. This fused lacZ'-IGF-II gene was under the control of tac promoter and we overproduced the beta-galactosidase-IGF-II fusion protein in the Escherichia coli. The fusion protein formed inclusion bodies inside the cells. The fusion protein was purified from the isolated inclusion bodies and IGF-II protein was obtained from their fusion protein by CNBr cleavage. The released IGF-II was confirmed by its molecular weight as determined by SDS-PAGE and by its ability to bind anti-IGF antibody.

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Biotechnology
  • Cloning, Molecular
  • DNA / genetics
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Humans
  • Insulin-Like Growth Factor II / biosynthesis*
  • Insulin-Like Growth Factor II / genetics
  • Insulin-Like Growth Factor II / isolation & purification
  • Molecular Sequence Data
  • Plasmids
  • Recombinant Fusion Proteins / biosynthesis
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / isolation & purification
  • Somatomedins / biosynthesis*

Substances

  • Recombinant Fusion Proteins
  • Somatomedins
  • Insulin-Like Growth Factor II
  • DNA