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Applied and Environmental Microbiology, April 2005, p. 2140-2144, Vol. 71, No. 4
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.4.2140-2144.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

SHORT REPORT

New Tool for Metabolic Pathway Engineering in Escherichia coli: One-Step Method To Modulate Expression of Chromosomal Genes

Isabelle Meynial-Salles,1 Marguerite A. Cervin,2 and Philippe Soucaille3*

Laboratoire de Biotechnologie-Bioprocédés, UMR INSA-CNRS 5504, UMR INSA-INRA 792, INSA,3 CRITT-Bioindustries CRT, INSA, DGBA, Toulouse, France,1 Genencor International, Palo Alto, California2

Received 30 June 2004/ Accepted 9 November 2004

A simple and highly efficient method was developed to produce a library of Escherichia coli clones that express a particular chromosomal gene at a wide range of expression levels. The basic strategy was to replace all or part of the upstream region of a coding sequence containing the elements involved in its expression (promoter, operator, gene coding for a regulator, ribosome binding site, and start codon) with a PCR-generated library of expression cassettes.

* Corresponding author. Mailing address: Laboratoire de Biotechnologie-Bioprocédés, UMR INSA-CNRS 5504, UMR INSA-INRA 792, INSA, 135 avenue de Rangueil, 31077 Toulouse cedex 4, France. Phone: (33) 5 61 55 94 52. Fax: (33) 5 61 55 94 00. E-mail: soucaille{at}insa-tlse.fr.

Applied and Environmental Microbiology, April 2005, p. 2140-2144, Vol. 71, No. 4
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.4.2140-2144.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

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