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Конъюгативных Спаривание Анализы для последовательности конкретного анализа переноса белков, участвующих в конъюгации бактерий
JoVE Journal
Genética
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JoVE Journal Genética
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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Capítulos

  • 00:05Título
  • 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

Tadução automática

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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