High level expression of soluble angiogenin in Escherichia coli

Biochem Mol Biol Int. 1999 Feb;47(2):267-73. doi: 10.1080/15216549900201283.

Abstract

Human angiogenin was genetically engineered and contained the E. coli Omp A signal sequence for secreting soluble angiogenin to the periplasm under tac promoter control. The angiogenin sequence was encoded in a single gene and expressed as a 14.4 kilodalton soluble protein in E. coli. It was purified by CM-Sepharose ion-exchange chromatography and by a heparin-Sepharose affinity chromatography procedure. The biological activity of angiogenin was established by its ability to inhibit mRNA-dependent rabbit reticulocyte cell-free translation.

MeSH terms

  • Bacterial Outer Membrane Proteins / genetics*
  • Chromatography, Affinity
  • Chromatography, Agarose
  • Escherichia coli / genetics*
  • Gene Expression
  • Genetic Engineering
  • Humans
  • Plasmids / genetics
  • Promoter Regions, Genetic
  • Protein Biosynthesis*
  • Proteins / genetics
  • Proteins / isolation & purification
  • Recombinant Proteins / isolation & purification
  • Reticulocytes / metabolism
  • Ribonuclease, Pancreatic*
  • Solubility

Substances

  • Bacterial Outer Membrane Proteins
  • Proteins
  • Recombinant Proteins
  • OMPA outer membrane proteins
  • angiogenin
  • Ribonuclease, Pancreatic