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.
Department of Biochemistry and Biophysics, Stockholm University, S-106 91 Stockholm, Sweden.
Tobias
T
Cornvik
Sue-Li
SL
Dahlroth
Audur
A
Magnusdottir
Maria Dolores
MD
Herman
Rosemarie
R
Knaust
Monica
M
Ekberg
Pär
P
Nordlund
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