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Use of Two-Dimensional Electrophoresis to Explore Foodborne Bacterial Pathogen Responses to Gastrointestinal Stress

  • Denise Schrama
  • Maria Leonor FaleiroEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1918)

Abstract

Proteomics was applied here to study Listeria monocytogenes response to gastrointestinal stress. It separated extracted proteins by their isoelectric point (pI) in the first dimension followed by separation by molecular weight in the second dimension on a polyacrylamide gel. L. monocytogenes was grown in an appropriate culture medium after which it was transferred to a simulated cheese medium for 2 h. Bacteria were exposed to gastric stress using artificial saliva and gastric fluid for 5 min and 2 h, respectively. After each step samples were taken for protein extraction and a two-dimensional electrophoresis approach. Proteins were separated on 18 cm Immobiline DryStrip gels with a pH range of 4–7 and the protein pattern analyzed.

Key words

Listeria monocytogenes Cheese medium Artificial saliva Gastric stress Proteomics 

Notes

Acknowledgments

This work was supported by Fundação para a Ciência e Tecnologia (PTDC/AGRI-ALI/2006 and IBB/CBME, LA, FEDER/POCI).

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.CCMARUniversity of AlgarveFaroPortugal
  2. 2.CMBRUniversity of AlgarveFaroPortugal

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