Scarless engineering of the Escherichia coli genome

Tamás Fehér, Ildikó Karcagi, Zsuzsa Gyorfy, Kinga Umenhoffer, Bálint Csörgo, György Pósfai

Research output: Chapter in Book/Report/Conference proceedingChapter

17 Citations (Scopus)

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.

Original languageEnglish
Title of host publicationMicrobial Gene Essentiality Protocols and Bioinformatics
Pages251-259
Number of pages9
Publication statusPublished - Dec 1 2007

Publication series

NameMethods in Molecular Biology
Volume416
ISSN (Print)1064-3745

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Keywords

  • Double-strand break
  • Escherichia coli
  • I-Scel endonuclease
  • Lambda Red recombinase
  • Recombination
  • Serial genome modification
  • Suicide plasmid

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Fehér, T., Karcagi, I., Gyorfy, Z., Umenhoffer, K., Csörgo, B., & Pósfai, G. (2007). Scarless engineering of the Escherichia coli genome. In Microbial Gene Essentiality Protocols and Bioinformatics (pp. 251-259). (Methods in Molecular Biology; Vol. 416).