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細菌接合に関与する転写タンパク質の配列特異的分析のための接合交配アッセイ
JoVE 杂志
遗传学
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JoVE 杂志 遗传学
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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Chapters

  • 00:05Title
  • 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

自动翻译

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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