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Genetic Manipulation of the Plant Pathogen Ustilago maydis to Study Fungal Biology and Plant Microbe Interactions
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Genetics
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JoVE Journal Genetics
Genetic Manipulation of the Plant Pathogen Ustilago maydis to Study Fungal Biology and Plant Microbe Interactions

Genetic Manipulation of the Plant Pathogen Ustilago maydis to Study Fungal Biology and Plant Microbe Interactions

DOI:
10.3791/54522-v

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11:42 min

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September 30, 2016

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, , , , ,
1Institute for Microbiology,Heinrich-Heine University Düsseldorf, 2Bioeconomy Science Center (BioSC), 3Department of Genetics, Institute of Applied Biosciences,Karlsruhe Institute of Technology, 4Cluster of Excellence in Plant Sciences (CEPLAS),Heinrich-Heine University Düsseldorf

Chapters

  • 00:05Title
  • 00:46Protoplast Preparation
  • 04:08Transformation of U. maydis
  • 06:35Verification of Deletion Events: Preparing gDNA
  • 09:17Results: Deletion Mutant Experiment
  • 10:43Conclusion

Summary

Automatic Translation

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We describe a robust gene replacement strategy to genetically manipulate the smut fungus Ustilago maydis. This protocol explains how to generate deletion mutants to investigate infection phenotypes. It can be extended to modify genes in any desired way, e.g., by adding a sequence encoding a fluorescent protein tag.

Tags

Genetic ManipulationUstilago MaydisFungal BiologyPlant-microbe InteractionsGene DeletionPromoter ExchangeFusion ConstructsProtoplastingTransformationCell CultureOptical DensityContaminationCentrifugationProtoplasting SolutionMicroscopy

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