Abstract
Lambda Red recombineering is an easy and efficient method for generating genetic modifications in Escherichia coli. For gene deletions, lambda Red recombineering is combined with the use of selectable markers, which are removed through the action of, e.g., flippase (Flp) recombinase. This PCR-based engineering method has also been applied to a number of other bacteria. In this chapter, we describe a recently developed one plasmid-based method as well as the use of a strain with genomically integrated recombineering genes, which significantly speeds up the engineering of strains with multiple genomic alterations.
| Original language | English |
|---|---|
| Title of host publication | Synthetic Metabolic Pathways |
| Volume | 1671 |
| Publication date | 2018 |
| Pages | 119-129 |
| ISBN (Print) | 978-1-4939-7294-4 |
| ISBN (Electronic) | 978-1-4939-7295-1 |
| DOIs | |
| Publication status | Published - 2018 |
| Series | Methods in Molecular Biology |
|---|---|
| ISSN | 1064-3745 |
Keywords
- Flippase recombinase
- Genome editing
- Industrial biotechnology
- Lambda Red recombineering
- Synthetic biology
- pSIJ8
Cite this
Ingemann Jensen, S., & Nielsen, A. T. (2018). Multiplex Genome Editing in Escherichia coli. In Synthetic Metabolic Pathways (Vol. 1671, pp. 119-129). Methods in Molecular Biology https://doi.org/10.1007/978-1-4939-7295-1_8