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Published Online April 27, 2006
Science DOI: 10.1126/science.1126439

Reports

Submitted on February 21, 2006
Accepted on April 19, 2006

Emergent Properties of Reduced-Genome Escherichia coli

György Pósfai 1*, Guy Plunkett III 2, Tamás Fehér 3, David Frisch 4, Günther M. Keil 5, Kinga Umenhoffer 3, Vitaliy Kolisnychenko 6, Buffy Stahl 7, Shamik S. Sharma 8, Monika de Arruda 7, Valerie Burland 9, Sarah W. Harcum 10, Frederick R. Blattner 2*

1 Institute of Biochemistry, Biological Research Center, H-6701 Szeged, Hungary; Scarab Genomics LLC, Madison, WI 53713, USA.
2 Scarab Genomics LLC, Madison, WI 53713, USA; Department of Genetics; Genome Center of Wisconsin, University of Wisconsin, Madison, WI 53706, USA.
3 Institute of Biochemistry, Biological Research Center, H-6701 Szeged, Hungary.
4 Scarab Genomics LLC, Madison, WI 53713, USA; Genome Center of Wisconsin, University of Wisconsin, Madison, WI 53706, USA.
5 Federal Research Centre for Virus Diseases of Animals, Institute of Molecular Biology, Friedrich-Loeffler Institutes, D-17493 Greifswald-Insel Riems, Germany.
6 Institute of Biochemistry, Biological Research Center, H-6701 Szeged, Hungary. Present address: National Heart, Lung, and Blood Institute, 9000 Rockville Pike, Building 10, Room 7D05, Bethesda, MD 20852, USA.
7 Scarab Genomics LLC, Madison, WI 53713, USA.
8 Department of Chemical and Biomolecular Engineering, 7Department of Bioengineering, Clemson University, Clemson, SC 29634, USA. Present address: New England Biolabs, 240 County Road, Ipswich, MA 01938, USA.
9 Scarab Genomics LLC, Madison, WI 53713, USA; Department of Genetics
10 Department of Bioengineering, Clemson University, Clemson, SC 29634, USA.

* To whom correspondence should be addressed.
György Pósfai , E-mail: posfaigy{at}nucleus.szbk.u-szeged.hu
Frederick R. Blattner , E-mail: fred{at}genome.wisc.edu

Using synthetic biology we reduced the Escherichia coli K-12 genome by making planned, precise deletions. The multiple-deletion series (MDS) strains, with genome reductions up to 15%, were designed by identifying non-essential genes and sequences for elimination, including recombinogenic or mobile DNA and cryptic virulence genes, while preserving good growth profiles and protein production. Genome reduction also led to unanticipated beneficial properties characterized here -- high electroporation efficiency and accurate propagation of recombinant genes and plasmids that were unstable in other strains. Eradication of stress-induced transposition has stabilized the MDS genomes and provides some of the new properties.


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