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Generation of Null Mutants to Elucidate the Role of Bacterial Glycosyltransferases in Bacterial Motility
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Immunologia e infezioni
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JoVE Journal Immunologia e infezioni
Generation of Null Mutants to Elucidate the Role of Bacterial Glycosyltransferases in Bacterial Motility

Generation of Null Mutants to Elucidate the Role of Bacterial Glycosyltransferases in Bacterial Motility

DOI:
10.3791/63231-v

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12:29 min

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March 11, 2022

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, , ,
1Departamento de Genética, Microbiología y Estadística,Universidad de Barcelona, 2Human Health Therapeutics Research Centre,National Research Council Canada, 3Department of Chemistry,Carleton University, 4Department of Biology,Carleton University

Capitoli

  • 00:04Introduction
  • 01:01Construction of pDM4 Recombinant Plasmid with PCR In-Frame Deletion Products
  • 03:10Preparation of Electrocompetent E. coli and Electroporation of pDM4 Recombinant Plasmid
  • 04:52Transfer of In-Frame Deletion to the Aeromonas Strain
  • 07:44Motility Assays in Liquid and Semi-Solid Media
  • 08:34Isolation and Purification of Flagella
  • 10:23Results: Confirmation of Null Mutants and Phenotypic Analysis of Polar Flagella
  • 11:47Conclusion

Summary

Traduzione automatica

Traduzione automatica

Here, we describe the construction of null mutants of Aeromonas in specific glycosyltransferases or regions containing glycosyltransferases, the motility assays, and flagella purification performed to establish the involvement and function of their encoded enzymes in the biosynthesis of a glycan, as well as the role of this glycan in bacterial pathogenesis.

Tags

Null MutantsBacterial GlycosyltransferasesBacterial MotilityFlagella GlycosylationIn-frame DeletionPCRCloningPDM4 VectorE. Coli Transformation

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