Amplification of the Chloramphenicol Acetyltransferase (CAT) Cassette from pBAD33 Plasmid Using Oligomers Designed for Homologous Pairing
2:38
Generation of pOX38-Tc Δgene::Cm Strains
4:26
Conjugative Mating Assay from XK1200 Donors to MC4100 Recipients
8:38
Results: Transfer Gene Knockout and Recovery of Conjugative DNA Transfer
9:45
Conclusion
Transcript
The overall goal of this mating assay is to assess the effects of mutations within the conjugative transfer gene on its ability to affect plasmid transfer within the context of a functional type four secretion complex. This method allows for askin
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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.