Back to Search
Start Over
Rapid oligonucleotide-based recombineering of the chromosome of Salmonella enterica.
- Source :
-
Applied and environmental microbiology [Appl Environ Microbiol] 2009 Mar; Vol. 75 (6), pp. 1575-80. Date of Electronic Publication: 2009 Jan 16. - Publication Year :
- 2009
-
Abstract
- Recombinant engineering using Red recombinase-based approaches offers efficient and rapid approaches to deletion and modification of genes. Here we describe a novel application of Red recombinant engineering that enables direct manipulation of chromosomal loci by electroporation with short synthetic DNA molecules. We demonstrate the use of this approach for the generation of scarless in-frame deletions in chromosomal genes of Salmonella enterica. Furthermore, we describe rapid site-directed mutagenesis within bacterial chromosomes without any requirement for cloning and mutating genes in vitro or for reintroducing mutant alleles into the chromosome. This approach can be expected to facilitate mutational analysis in S. enterica and in other bacterial species able to support Red-mediated recombination.
Details
- Language :
- English
- ISSN :
- 1098-5336
- Volume :
- 75
- Issue :
- 6
- Database :
- MEDLINE
- Journal :
- Applied and environmental microbiology
- Publication Type :
- Academic Journal
- Accession number :
- 19151186
- Full Text :
- https://doi.org/10.1128/AEM.02509-08