Journal article · Book chapter
Multiplex Genome Editing in Escherichia coli
Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark1
Bacterial Cell Factory Optimization, Research Groups, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark2
Research Groups, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark3
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.
| Language: | English |
|---|---|
| Year: | 2018 |
| Pages: | 119-129 |
| Series: | Methods in Molecular Biology |
| ISBN: | 1493972944 , 1493972952 , 9781493972944 and 9781493972951 |
| ISSN: | 19406029 and 10643745 |
| Types: | Journal article and Book chapter |
| DOI: | 10.1007/978-1-4939-7295-1_8 |
| ORCIDs: | Ingemann Jensen, Sheila and Nielsen, Alex Toftgaard |
