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Lee, Sung Kuk
Synthetic Biology & Metabolic Engineering Laboratory
Research Interests
  • Biofuels production
  • Gene discovery
  • Protein engineering
  • Development of gene expression systems
  • Metabolic pathway regulation
  • Metabolic pathway optimization using functional genomics
  • Synthetic biology
  • Development of biosensors

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A Simple and Effective Method for Construction of Escherichia coli Strains Proficient for Genome Engineering

Cited 7 times inthomson ciCited 5 times inthomson ci
Title
A Simple and Effective Method for Construction of Escherichia coli Strains Proficient for Genome Engineering
Author
Ryu, Young ShinBiswas, Rajesh KumarShin, KwangsuParisutham, VinuselviKim, Suk MinLee, Sung Kuk
Issue Date
2014-04
Publisher
PUBLIC LIBRARY SCIENCE
Citation
PLOS ONE, v.9, no.4, pp.e94266
Abstract
Multiplex genome engineering is a standalone recombineering tool for large-scale programming and accelerated evolution of cells. However, this advanced genome engineering technique has been limited to use in selected bacterial strains. We developed a simple and effective strain-independent method for effective genome engineering in Escherichia coli. The method involves introducing a suicide plasmid carrying the l Red recombination system into the mutS gene. The suicide plasmid can be excised from the chromosome via selection in the absence of antibiotics, thus allowing transient inactivation of the mismatch repair system during genome engineering. In addition, we developed another suicide plasmid that enables integration of large DNA fragments into the lacZ genomic locus. These features enable this system to be applied in the exploitation of the benefits of genome engineering in synthetic biology, as well as the metabolic engineering of different strains of E. coli.
URI
https://scholarworks.unist.ac.kr/handle/201301/10090
DOI
10.1371/journal.pone.0094266
ISSN
1932-6203
Appears in Collections:
ECHE_Journal Papers
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