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Geração de Mutantes Nulos para Elucidar o Papel das Glicosyltransferases Bacterianas na Motilidade Bacteriana
JoVE Journal
Immunology and Infection
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JoVE Journal Immunology and Infection
Generation of Null Mutants to Elucidate the Role of Bacterial Glycosyltransferases in Bacterial Motility
DOI:

12:29 min

March 11, 2022

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Chapters

  • 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

Automatic Translation

Aqui, descrevemos a construção de mutantes nulos de Aeromonas em glicosyltransferases específicas ou regiões contendo glicosyltransferases, os ensaios de motilidade e a purificação de flagela realizadas para estabelecer o envolvimento e função de suas enzimas codificadas na biossíntese de um glicano, bem como o papel deste glicano em patógenos bacterianos.

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