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Title: Electroporation of Functional Bacterial Effectors into Mammalian Cells

Abstract

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.

Authors:
 [1];  [2];  [2];  [3];  [3];  [3];  [1];  [1];  [4]
  1. Pacific Northwest National Lab. (PNNL), Richland, WA (United States). Biological Sciences Division
  2. Pacific Northwest National Lab. (PNNL), Richland, WA (United States). Environmental Molecular Science Lab.
  3. Univ. of Toronto, ON (Canada). Ontario Center for Structural Proteomics
  4. Washington State Univ., Pullman, WA (United States). Center for Bioproducts and Bioenergy
Publication Date:
Research Org.:
Pacific Northwest National Lab. (PNNL), Richland, WA (United States). Environmental Molecular Sciences Lab. (EMSL)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)
OSTI Identifier:
1184956
Report Number(s):
PNNL-SA-102481
Journal ID: ISSN 1940-087X; jove; 42696; 400412000
Grant/Contract Number:  
AC05-76RL01830
Resource Type:
Accepted Manuscript
Journal Name:
Journal of Visualized Experiments
Additional Journal Information:
Journal Issue: 95; Journal ID: ISSN 1940-087X
Publisher:
MyJoVE Corp.
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; electroporation; protein; transfection; expression; localization; confocal microscopy; Salmonella; effector; Immunology; Issue 95; Environmental Molecular Sciences Laboratory

Citation Formats

Sontag, Ryan L., Mihai, Cosmin, Orr, Galya, Savchenko, Alexei, Skarina, Tatiana, Cui, Hong, Cort, John R., Adkins, Joshua N., and Brown, Roslyn N. Electroporation of Functional Bacterial Effectors into Mammalian Cells. United States: N. p., 2015. Web. doi:10.3791/52296.
Sontag, Ryan L., Mihai, Cosmin, Orr, Galya, Savchenko, Alexei, Skarina, Tatiana, Cui, Hong, Cort, John R., Adkins, Joshua N., & Brown, Roslyn N. Electroporation of Functional Bacterial Effectors into Mammalian Cells. United States. doi:10.3791/52296.
Sontag, Ryan L., Mihai, Cosmin, Orr, Galya, Savchenko, Alexei, Skarina, Tatiana, Cui, Hong, Cort, John R., Adkins, Joshua N., and Brown, Roslyn N. Mon . "Electroporation of Functional Bacterial Effectors into Mammalian Cells". United States. doi:10.3791/52296. https://www.osti.gov/servlets/purl/1184956.
@article{osti_1184956,
title = {Electroporation of Functional Bacterial Effectors into Mammalian Cells},
author = {Sontag, Ryan L. and Mihai, Cosmin and Orr, Galya and Savchenko, Alexei and Skarina, Tatiana and Cui, Hong and Cort, John R. and Adkins, Joshua N. and Brown, Roslyn N.},
abstractNote = {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.},
doi = {10.3791/52296},
journal = {Journal of Visualized Experiments},
number = 95,
volume = ,
place = {United States},
year = {2015},
month = {1}
}

Journal Article:
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