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Conjugatif Mating Assays pour l'analyse des protéines de transfert impliqués dans Conjugaison bactérienne spécifique à la séquence
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Génétique
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Journal JoVE Génétique
Conjugative Mating Assays for Sequence-specific Analysis of Transfer Proteins Involved in Bacterial Conjugation
DOI:

10:41 min

January 04, 2017

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Chapitres

  • 00:05Titre
  • 00:59Amplification of the Chloramphenicol Acetyltransferase (CAT) Cassette from pBAD33 Plasmid Using Oligomers Designed for Homologous Pairing
  • 02:38Generation of pOX38-Tc Δgene::Cm Strains
  • 04:26Conjugative Mating Assay from XK1200 Donors to MC4100 Recipients
  • 08:38Results: Transfer Gene Knockout and Recovery of Conjugative DNA Transfer
  • 09:45Conclusion

Summary

Traduction automatique

Here, we present a protocol to knockout a gene of interest involved in plasmid conjugation and subsequently analyze the impact of its absence using mating assays. The function of the gene is further explored to a specific region of its sequence using deletion or point mutations.

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