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 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.
Cornvik
Tobias
T
Department of Biochemistry and Biophysics, Stockholm University, S-106 91 Stockholm, Sweden.
Dahlroth
Sue-Li
SL
Magnusdottir
Audur
A
Herman
Maria Dolores
MD
Knaust
Rosemarie
R
Ekberg
Monica
M
Nordlund
Pär
P
D018929:Cell Culture Techniques / Q000379:methods*; D029968:Escherichia coli Proteins / Q000302:isolation & purification* / Q000378:metabolism*; D020869:Gene Expression Profiling / Q000379:methods*; D002479:Inclusion Bodies / Q000737:chemistry* / Q000378:metabolism*; D012995:Solubility; D014462:Ultrafiltration / Q000379:methods*
D029968:Escherichia coli Proteins