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Applied and Environmental Microbiology, June 2009, p. 3803-3807, Vol. 75, No. 11
0099-2240/09/$08.00+0     doi:10.1128/AEM.00255-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Production of Recombinant Proteins in the lon-Deficient BL21(DE3) Strain of Escherichia coli in the Absence of the DnaK Chaperone{triangledown} ,{dagger}

Julien Ratelade,1 Marie-Caroline Miot,2 Emmett Johnson,3 Jean-Michel Betton,2 Philippe Mazodier,4 and Nadia Benaroudj4*

Unité de Nephropathies Héréditaires et Rein en Développement, INSERM U574, Hôpital Necker-Enfants Malades, 149 Rue de Sèvres, 75743 Paris Cedex 15, France,1 Institut Pasteur, Unité de Biochimie Structurale, CNRS URA 2185, F-75015 Paris, France,2 Department of Microbiology and Immunology, Tulane Medical School, 1430 Tulane Avenue, New Orleans, Louisiana 70112,3 Institut Pasteur, Unité de Biologie des Bactéries Pathogènes à Gram-Positif, CNRS URA 2172, F-75015 Paris, France4

Received 2 February 2009/ Accepted 24 March 2009

To eliminate unavoidable contamination of purified recombinant proteins by DnaK, we present a unique approach employing a BL21(DE3) {Delta}dnaK 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.

* Corresponding author. Mailing address: Institut Pasteur, 25 rue du Dr Roux, 75015 Paris, France. Phone: 33 (0)1 45 68 88 12. Fax: 33 (0)1 45 68 89 38. E-mail: nadia.benaroudj{at}pasteur.fr

{triangledown} Published ahead of print on 3 April 2009.

{dagger} Supplemental material for this article may be found at http://aem.asm.org/.

Applied and Environmental Microbiology, June 2009, p. 3803-3807, Vol. 75, No. 11
0099-2240/09/$08.00+0     doi:10.1128/AEM.00255-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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