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TransFLP — A Method to Genetically Modify Vibrio cholerae Based on Natural Transformation and FLP-recombination
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Chapters
- 00:05Title
- 01:16Preparation of Chitin Flakes
- 01:39Oligonucleotide Design and Polymerase Chain Reaction
- 03:45Chitin-induced Natural Transformation
- 04:59Removal of Selective Cassette(s) by FLP-recombination
- 05:46Plasmid Curing
- 06:21Results: Representative TransFLP Modifications of V. cholerae
- 11:34Conclusion
A quick method to modify the genome of V. cholerae is described. These modifications include the deletion of single genes, gene clusters and genomic islands as well as the integration of short sequences (e.g. promoter elements or affinity-tag sequences). The method is based on the natural transformation and FLP-recombination.
Tags
TransFLPGenetically ModifyVibrio CholeraeNatural TransformationFLP-recombinationBacterial ChromosomesReplicative PlasmidsHomology-mediated RecombinationInsertional MutantsPlasmid ExcisionGram-negative BacteriaLevan Sucrase EnzymeSacB GenePre-insertion GenotypeModified GeneTime-consumingCloningHorizontal TransferArtificial TransformationPositive SelectionRapid ManipulationChitin-mediated InductionNatural Competence
