E. coli-based in vitro transcription/translation: in vivo-specific synthesis rates and high yields in a batch system

Biotechniques. 1998 May;24(5):862-8. doi: 10.2144/98245rr03.

Abstract

A highly efficient Escherichia coli-based, batch in vitro protein synthesis system using circular plasmid DNA is described. Compared to a presently available commercial kit, this improved system produced several hundredfold greater yields of the rDNA human protein thrombopoietin (ca. 450 micrograms/mL). The system is capable of obtaining specific synthesis rates similar to those in vivo, approximately a 1000-fold increase compared to the original methods previously described. It compares favorably in rates and yields to the recently published semicontinuous methods but with the convenience of a true batch system.

MeSH terms

  • Bacterial Proteins / biosynthesis
  • Cell Fractionation
  • DNA, Bacterial / isolation & purification
  • Electrophoresis, Polyacrylamide Gel
  • Escherichia coli / genetics*
  • Humans
  • Plasmids / genetics
  • Protein Biosynthesis*
  • Recombinant Proteins / biosynthesis*
  • Sodium Dodecyl Sulfate
  • Templates, Genetic
  • Thrombopoietin / biosynthesis
  • Thrombopoietin / genetics
  • Transcription, Genetic*

Substances

  • Bacterial Proteins
  • DNA, Bacterial
  • Recombinant Proteins
  • Sodium Dodecyl Sulfate
  • Thrombopoietin