Production of recombinant proteins in the lon-deficient BL21(DE3) strain of Escherichia coli in the absence of the DnaK chaperone.
To eliminate unavoidable contamination of purified recombinant proteins by DnaK, we present a unique approach employing a BL21(DE3) DeltadnaK strain of Escherichia coli. Selected representative purified proteins remained soluble, correctly assembled, and active. This finding establishes DnaK dispensability for protein production in BL21(DE3), which is void of Lon protease, key to eliminating unfolded proteins.
Ratelade
Julien
J
INSERM U, Hôpital Necker-Enfants Malades, Paris, France.
Miot
Marie-Caroline
MC
Johnson
Emmett
E
Betton
Jean-Michel
JM
Mazodier
Philippe
P
Benaroudj
Nadia
N
D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins; D017353:Gene Deletion; D018840:HSP70 Heat-Shock Proteins / Q000172:deficiency*; D011994:Recombinant Proteins / Q000096:biosynthesis*
D029968:Escherichia coli Proteins; D018840:HSP70 Heat-Shock Proteins; D011994:Recombinant Proteins; C063842:dnaK protein, E coli