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Site-specific Bacterial Chromosome Engineering: ΦC31 Integrase Mediated Cassette Exchange (IMCE)
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Chapters
- 00:05Title
- 01:46Production of Culture
- 02:53Culture Preparation and Mixing (Mating)
- 04:27Isolation of Trans-integrants
- 05:55Results of Integrase Mediated Cassette Exchange (IMCE)
- 08:01Conclusion
A quick and efficient method to integrate foreign DNA of interest into pre-made acceptor strains, termed landing pad strains, is described. The method allows site-specific integration of a DNA cassette into the engineered landing pad locus of a given strain, through conjugation and expression of the ΦC31 integrase.
Tags
Site-specificBacterial Chromosome EngineeringΦC31 IntegraseCassette ExchangeAttBAttPRecombinationLanding PadSpectinomycin-resistance GeneAadAE. Coli ß-glucuronidase GeneUidASinorhizobium MelilotiOchrobactrum AnthropiAgrobacterium TumefaciensAmpC LocusTetA LocusMobilizable Donor VectorsStuffer Red Fluorescent Protein (rfp) GeneAntibiotic Resistance GeneGentamicin Resistant Plasmid PJH110SphIPstI
