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

Journal

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免疫与感染

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电功能性细菌效应器到哺乳动物细胞

Electroporation of Functional Bacterial Effectors into Mammalian Cells

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免疫与感染

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JoVE 杂志免疫与感染

Electroporation of Functional Bacterial Effectors into Mammalian Cells

电功能性细菌效应器到哺乳动物细胞

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10,058 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

•

10015 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

Summary

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.