Colony filtration blot: a new screening method for soluble protein expression in Escherichia coli
- 22 June 2005
- journal article
- research article
- Published by Springer Science and Business Media LLC in Nature Methods
- Vol. 2 (7), 507-509
- https://doi.org/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.Keywords
This publication has 16 references indexed in Scilit:
- Genetic screens and directed evolution for protein solubilityCurrent Opinion in Chemical Biology, 2003
- Protein expression systems for structural genomics and proteomicsCurrent Opinion in Chemical Biology, 2003
- Proteome-scale purification of human proteins from bacteriaProceedings of the National Academy of Sciences of the United States of America, 2002
- Rapid screening for improved solubility of small human proteins produced as fusion proteins in Escherichia coliProtein Science, 2002
- Screening for Soluble Expression of Recombinant Proteins in a 96-Well FormatAnalytical Biochemistry, 2001
- Protein solubility and folding monitored in vivo by structural complementation of a genetic marker proteinNature Biotechnology, 2001
- Protein production: feeding the crystallographers and NMR spectroscopists.Nature Structural & Molecular Biology, 2000
- Rapid protein-folding assay using green fluorescent proteinNature Biotechnology, 1999
- Properties of Soluble Fusions Between Mammalian Aspartic Proteinases and Bacterial Maltose-Binding ProteinProtein Journal, 1999
- A simple in vivo assay for increased protein solubilityProtein Science, 1999