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Producing Gene Deletions in Escherichia coli by P1 Transduction with Excisable Antibiotic Resistance Cassettes
Journal JoVE
Génétique
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Journal JoVE Génétique
Producing Gene Deletions in Escherichia coli by P1 Transduction with Excisable Antibiotic Resistance Cassettes
DOI:

08:13 min

September 01, 2018

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Chapitres

  • 00:04Titre
  • 00:28Phage Lysate Preparation
  • 02:31P1 Transduction
  • 04:30Kanamycin (Kan) Cassette Excision
  • 06:03Results: TAA Expression by a tamA Deletion Strain
  • 07:38Conclusion

Summary

Traduction automatique

Here we present a protocol for the use of pre-existing antibiotic resistance-cassette deletion constructs as a basis for making deletion mutants in other E. coli strains. Such deletion mutations can be mobilized and inserted into the corresponding locus of a recipient strain using P1 bacteriophage transduction.

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