An efficient cell-free protein synthesis system using periplasmic phosphatase-removed S30 extract

J Microbiol Methods. 2000 Dec 15;43(2):91-6. doi: 10.1016/s0167-7012(00)00206-2.

Abstract

An efficient cell-free translation system was developed by removal of phosphatase localized in the periplasmic space, which hampers the translation reaction by hydrolyzing ATP. S30 extract was prepared from the spheroplast of Escherichia coli, and as much as 40% of ATP-hydrolysis activity of phosphatases could be easily removed by the spheroplast formation. The reaction period of translation using phosphatase-removed S30 extract could be prolonged and protein synthesis was enhanced by more than 30%.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell-Free System*
  • Escherichia coli / metabolism*
  • Periplasm / enzymology
  • Phosphoric Monoester Hydrolases / metabolism
  • Protein Biosynthesis*
  • Spheroplasts / metabolism
  • Subcellular Fractions / metabolism

Substances

  • Phosphoric Monoester Hydrolases