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Immunology and Infection

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Nanyang Technological University

TransFLP — A Method to Genetically Modify Vibrio cholerae Based on Natural Transformation and FLP-recombination

14.6K Views

•

12:13 min

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October 8th, 2012

DOI :

October 8th, 2012

•

Melanie Blokesch¹

¹Global Health Institute, School of Life Sciences, Ecole Polytechnique Fédérale de Lausanne (EPFL)

Summary

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.

Chapters in this video

0:05

Title

1:16

Preparation of Chitin Flakes

1:39

Oligonucleotide Design and Polymerase Chain Reaction

3:45

Chitin-induced Natural Transformation

4:59

Removal of Selective Cassette(s) by FLP-recombination

5:46

Plasmid Curing

6:21

Results: Representative TransFLP Modifications of V. cholerae

11:34

Conclusion

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