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

Juan M. Tom&#225;s; Kelly M. Fulton; Susan M. Twine; Susana Merino

doi:10.3791/63231

Journal

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Immunology and Infection

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产生空突变体以阐明细菌糖基转移酶在细菌运动中的作用

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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

Please note that all translations are automatically generated. Click here for the English version.

产生空突变体以阐明细菌糖基转移酶在细菌运动中的作用

DOI:

10.3791/63231-v

•

12:29 min

•

March 11, 2022

•

Juan M. Tomás, Kelly M. Fulton³, Susan M. Twine⁴, Susana Merino

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

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

在这里，我们描述了在特定的糖基转移酶或含有糖基转移酶的区域中 构建气单胞菌 的零突变体，进行运动性测定和鞭毛纯化以确定其编码酶在聚糖生物合成中的参与和功能，以及该聚糖在细菌发病机制中的作用。

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