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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

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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