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JoVE Journal Genetics
Conjugative Mating Assays for Sequence-specific Analysis of Transfer Proteins Involved in Bacterial Conjugation
 

Conjugative Mating Assays for Sequence-specific Analysis of Transfer Proteins Involved in Bacterial Conjugation

Article DOI: 10.3791/54854-v • 10:41 min • January 4th, 2017
January 4th, 2017 •
1, 1, 1, 1, 1, 1,2
1Department of Chemistry, York University, 2The Centre for Research on Biomolecular Interactions, York University

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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.

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Conjugative Mating Assays Sequence-specific Analysis Transfer Proteins Bacterial Conjugation Mutations Plasmid Transfer Type Four Secretion Complex Protein-protein Interactions Homologous Recombination Gene Function Knock-outs PBAD33 Plasmid Agarose Gel UV Exposure DNA Extraction Polymerase Chain Reaction (PCR) Reaction Mixture
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