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Producing Gene Deletions in Escherichia coli by P1 Transduction with Excisable Antibiotic Resistance Cassettes
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JoVE Journal Genetics
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:
10.3791/58267-v

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08:13 min

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September 01, 2018

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, ,
1Evolution and Genetics, Department of Biosciences,University of Oslo

Chapters

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

Summary

Automatic Translation

Automatic Translation

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.

Tags

Keywords: P1 TransductionGene DeletionEscherichia ColiAntibiotic Resistance CassetteKnockout MutantPhageBacterial CultureLysogeny BrothCentrifugationChloroformSodium Citrate

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