Colony filtration blot: a new screening method for soluble protein expression in Escherichia coli.
basic_science · Level V
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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.
Medical subject headings
- Cell Culture Techniques
- Escherichia coli Proteins
- Gene Expression Profiling
- Inclusion Bodies
- Ultrafiltration