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TransFLP - A基因修改方法<em>霍乱弧菌</em基于自然转化和FLP重组
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Immunology and Infection
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JoVE Journal Immunology and Infection
TransFLP — A Method to Genetically Modify Vibrio cholerae Based on Natural Transformation and FLP-recombination
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TransFLP - A基因修改方法<em>霍乱弧菌</em基于自然转化和FLP重组

DOI:
10.3791/3761-v

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12:13 min

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October 08, 2012

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1Global Health Institute, School of Life Sciences,Ecole Polytechnique Fédérale de Lausanne (EPFL)

Chapters

  • 00:05Title
  • 01:16Preparation of Chitin Flakes
  • 01:39Oligonucleotide Design and Polymerase Chain Reaction
  • 03:45Chitin-induced Natural Transformation
  • 04:59Removal of Selective Cassette(s) by FLP-recombination
  • 05:46Plasmid Curing
  • 06:21Results: Representative TransFLP Modifications of V. cholerae
  • 11:34Conclusion

Summary

Automatic Translation

Automatic Translation

一个快速的方法来修改的基因组<em> V.霍乱</em>进行说明。这些修改包括删除单个基因，基因簇和基因组岛屿以及集成的短序列（如启动子元件或亲和标签序列）。该方法是基于对自然的改造和FLP重组。

Tags

TransFLPGenetically ModifyVibrio CholeraeNatural TransformationFLP-recombinationBacterial ChromosomesReplicative PlasmidsHomology-mediated RecombinationInsertional MutantsPlasmid ExcisionGram-negative BacteriaLevan Sucrase EnzymeSacB GenePre-insertion GenotypeModified GeneTime-consumingCloningHorizontal TransferArtificial TransformationPositive SelectionRapid ManipulationChitin-mediated InductionNatural Competence

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