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

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 separat...

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Published inNature methods Vol. 2; no. 7; pp. 507 - 509
Main Authors Ekberg, Monica, Nordlund, Pär, Cornvik, Tobias, Dahlroth, Sue-Li, Magnusdottir, Audur, Herman, Maria Dolores, Knaust, Rosemarie
Format Journal Article
LanguageEnglish
Published United States Nature Publishing Group 01.07.2005
Subjects
Cell Culture Techniques - methods
E coli
Escherichia coli
Escherichia coli Proteins - isolation & purification
Escherichia coli Proteins - metabolism
Filtration
Gene Expression Profiling - methods
Inclusion Bodies - chemistry
Inclusion Bodies - metabolism
Mammals
Medicin och hälsovetenskap
Methods
Physiological aspects
Proteins
Solubility
Ultrafiltration - methods
Sweden
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Summary: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.
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ISSN:1548-7091
1548-7105
DOI:10.1038/nmeth767