title: Scarless engineering of the Escherichia coli genome.
abstract: E. coli K-12, being one of the best understood and thoroughly analyzed organisms, is the workhorse of genetic, biochemical, and systems biology research, as well as the platform of choice for numerous biotechnological applications. Genome minimization/remodeling is now a feasible approach to further enhance its beneficial characteristics for practical applications. Two genome engineering techniques, a lambda Red-mediated deletion method and a suicide (conditionally replicative) plasmid-based allele replacement procedure are presented here. These techniques utilize homologous recombination, and allow the rapid introduction of virtually any modifications in the genome.
authors.affiliation: Institute of Biochemistry, Biological Research Center of the Hungarian Academy of Sciences, Szeged, Hungary.
authors.forename: Tamás
authors.initials: T
authors.lastname: Fehér
authors.forename: Ildikó
authors.initials: I
authors.lastname: Karcagi
authors.forename: Zsuzsa
authors.initials: Z
authors.lastname: Gyorfy
authors.forename: Kinga
authors.initials: K
authors.lastname: Umenhoffer
authors.forename: Bálint
authors.initials: B
authors.lastname: Csörgo
authors.forename: György
authors.initials: G
authors.lastname: Pósfai
mesh_terms: D010582:Bacteriophage lambda / Q000235:genetics*; D001665:Binding Sites; D004269:DNA, Bacterial / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics*; D017353:Gene Deletion; D005818:Genetic Engineering / Q000379:methods*; D016680:Genome, Bacterial; D010957:Plasmids / Q000235:genetics; D011995:Recombination, Genetic
chemical_list: D004269:DNA, Bacterial