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Generation of Null Mutants to Elucidate the Role of Bacterial Glycosyltransferases in Bacterial Motility
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JoVE Journal 免疫学と感染
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, 3化学科,Carleton University, 4生物学科,Carleton University

章

  • 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

概要

自動翻訳

自動翻訳

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.

タグ

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

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