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Genetic Engineering of Dictyostelium discoideum Cells Based on Selection and Growth on Bacteria
Journal JoVE
Génétique
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Journal JoVE Génétique
Genetic Engineering of Dictyostelium discoideum Cells Based on Selection and Growth on Bacteria

Genetic Engineering of Dictyostelium discoideum Cells Based on Selection and Growth on Bacteria

DOI:
10.3791/58981-v

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06:08 min

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January 25, 2019

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, , , , , , ,
1MRC Laboratory of Molecular Biology, 2Department of Molecular and Cell Biology,University of Connecticut, 3Cancer Research UK Beatson Institute Glasgow, 4MRC Laboratory for Molecular Cell Biology,University College London, 5Department of Cell and Developmental Biology,University College London

Chapitres

  • 00:04Titre
  • 00:54Preparation of Cells and Materials
  • 02:30Transfection of Dictyostelium Cells Based on Bacterial Selectionin Protocol
  • 04:15Results: Dual Reporter Extrachromosomal Plasmid System Demonstrated
  • 05:37Conclusion

Summary

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Dictyostelium discoideum is a popular model organism to study complex cellular processes such as cell migration, endocytosis, and development. The utility of the organism is dependent on the feasibility of genetic manipulation. Here, we present methods to transfect Dictyostelium discoideum cells that overcome existing limitations of culturing cells in liquid media.

Tags

Dictyostelium DiscoideumGenetic EngineeringBacterial SelectionGrowth On BacteriaExochromosomal Plasmid TransfectionMicroscopyKlebsiella AerogenesSorensen BufferLB MediumOptical DensityH40 Electroporation BufferSM MediumAgar PlateCell TransferCentrifugationDNA Transformation

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