Colony filtration blot: a new screening method for soluble protein expression in Escherichia coli

Nat Methods. 2005 Jul;2(7):507-9. doi: 10.1038/nmeth767.

Abstract

The implementation of efficient technologies for the production of recombinant mammalian proteins remains an outstanding challenge in many structural and functional genomics programs. We have developed a new method for rapid identification of soluble protein expression in E. coli, based on a separation of soluble protein from inclusion bodies by a filtration step at the colony level. The colony filtration (CoFi) blot is very well suited to screen libraries, and in the present work we used it to screen a deletion mutagenesis library.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Culture Techniques / methods*
  • Escherichia coli Proteins / isolation & purification*
  • Escherichia coli Proteins / metabolism*
  • Gene Expression Profiling / methods*
  • Inclusion Bodies / chemistry*
  • Inclusion Bodies / metabolism*
  • Solubility
  • Ultrafiltration / methods*

Substances

  • Escherichia coli Proteins