Electroporation of Functional Bacterial Effectors into Mammalian Cells

Ryan L. Sontag; Cosmin Mihai; Galya Orr; Alexei Savchenko; Tatiana Skarina; Hong Cui; John R. Cort; Joshua N. Adkins; Roslyn N. Brown

doi:10.3791/52296

Журнал

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Иммунология и инфекции

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Электропорация функциональных бактериальных эффекторов в клетках млекопитающих

Electroporation of Functional Bacterial Effectors into Mammalian Cells

JoVE Journal
Иммунология и инфекции

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JoVE Journal Иммунология и инфекции

Electroporation of Functional Bacterial Effectors into Mammalian Cells

Электропорация функциональных бактериальных эффекторов в клетках млекопитающих

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

9,977 Views

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08:39 min

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January 19, 2015

DOI:

10.3791/52296-v

DOI:

10.3791/52296-v

•

08:39 min

January 19, 2015

•

2 Views

Ryan L. Sontag¹, Cosmin Mihai², Galya Orr², Alexei Savchenko³, Tatiana Skarina³, Hong Cui³, John R. Cort¹, Joshua N. Adkins¹, Roslyn N. Brown⁴

¹Biological Sciences Division,Pacific Northwest National Laboratory, ²Environmental Molecular Science Laboratory,Pacific Northwest National Laboratory, ³Structural Proteomics Group, Ontario Center for Structural Proteomics,University of Toronto, ⁴Center for Bioproducts and Bioenergy,Washington State University

Резюме

Electroporation was used to insert purified bacterial virulence effector proteins directly into living eukaryotic cells. Protein localization was monitored by confocal immunofluorescence microscopy. This method allows for studies on trafficking, function, and protein-protein interactions using active exogenous proteins, avoiding the need for heterologous expression in eukaryotic cells.