Abstract
Pseudomonas aeruginosa is a Gram negative bacterium. Separating the cell envelope compartments enables proteins to be localized to confirm where in the cell they function. Cell fractionation can also provide a first step in a protein purification strategy (Williams et al., 1998). This protocol has been designed to obtain the different fractions of P. aeruginosa, namely the inner membrane, outer membrane, cytoplasmic and periplasmic compartments. Specific detection of the arginine specific autotransporter (AaaA) (Luckett et al., 2012) in the outer membrane of P. aeruginosa has been performed using this protocol.
Materials and Reagents
Equipment
Procedure
Recipes
Acknowledgments
We acknowledge the use of this protocol in Luckett et al. (2012). We would like to thank the Chilean Government for financially supporting Esteban Paredes. We also thank Prof Miguel Camara for critical analysis of our work and everyone else in the Bacteriology Laboratories of the Centre of Biomolecular Sciences, University of Nottingham for helpful discussion about our work.
References
If you have any questions/comments about this protocol, you are highly recommended to post here. We will invite the authors of this protocol as well as some of its users to address your questions/comments. To make it easier for them to help you, you are encouraged to post your data including images for the troubleshooting.
Dear Marie,It depends what your final sample will be used for. The French Press lyses in a different way to sonication, and is better for bigger volumes as the sample will not warm up. It is worth trying another lysis if the French press is not available to see if it works for you, Regards, Kim