Emergent Properties of Reduced-Genome Escherichia coli
György Pósfai,1,2*
Guy Plunkett, III,2,3,4
Tamás Fehér,1
David Frisch,2,4
Günther M. Keil,5
Kinga Umenhoffer,1
Vitaliy Kolisnychenko,1
Buffy Stahl,2
Shamik S. Sharma,6
Monika de Arruda,2
Valerie Burland,2,3
Sarah W. Harcum,7
Frederick R. Blattner2,3,4*
With the use of 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 nonessential 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: high electroporation efficiency and accurate propagation of recombinant genes and plasmids that were unstable in other strains. Eradication of stress-induced transposition evidently stabilized the MDS genomes and provided some of the new properties.
1 Institute of Biochemistry, Biological Research Center, H-6726 Szeged, Hungary.
2 Scarab Genomics LLC, Madison, WI 53713, USA.
3 Department of Genetics, University of Wisconsin, Madison, WI 53706, USA.
4 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 Department of Chemical and Biomolecular Engineering, Clemson University, Clemson, SC 29634, USA.
7 Department of Bioengineering, Clemson University, Clemson, SC 29634, USA.
Present address: National Heart, Lung, and Blood Institute, 9000 Rockville Pike, Building 10, Room 7D05, Bethesda, MD 20852, USA.
Present address: New England Biolabs, 240 County Road, Ipswich, MA 01938, USA.
* To whom correspondence should be addressed. E-mail: fred{at}genome.wisc.edu (F.R.B.); posfaigy{at}nucleus.szbk.u-szeged.hu (G.P.)
