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An Electroporation Method to Transform Rickettsia spp. with a Fluorescent Protein-Expressing Shuttle Vector in Tick Cell Lines
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Biologia
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JoVE Journal Biologia
An Electroporation Method to Transform Rickettsia spp. with a Fluorescent Protein-Expressing Shuttle Vector in Tick Cell Lines

An Electroporation Method to Transform Rickettsia spp. with a Fluorescent Protein-Expressing Shuttle Vector in Tick Cell Lines

DOI:
10.3791/64562-v

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07:18 min

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October 11, 2022

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, , ,
1Department of Entomology,University of Minnesota

Capitoli

  • 00:04Introduction
  • 00:41Determining the Rickettsia parkeri Infection Rate via Geimsa Staining
  • 01:35Purification of Rickettsia parkeri from Tick Cell Culture
  • 03:35Transformation of Rickettsia parkeri with the pRAM18dSFA Plasmid
  • 05:52Results: Transformants of Rickettsia parkeri Expressing RFP in ISE6 Cells
  • 06:29Conclusion

Summary

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Electroporation is a rapid, broadly adopted method for introducing exogenous DNA into the genus Rickettsia. This protocol provides a useful electroporation method for the transformation of obligate intracellular bacteria in the genus Rickettsia.

Tags

Keywords: RickettsiaElectroporationFluorescent ProteinShuttle VectorTick Cell LinesGiemsa StainingCell-free RickettsiaSilicon Carbide GritCell-free PreparationSyringe Filter

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