Which type of assay can perform for determination of membrane integrity of gram-negative bacteria?
Dear Ashraf,
you can test the sensitivity of your bacteria to different antibiotic families. If your bacteria are sensitive to all, this means that their membrane is damaged. We use vancomycin,erythomycin, rifampicin, bacitracine, novobiocin. If we take the example of vancomycin : Gram negative bacteria are normally resistant to vancomycin. This is a large molecule that cannot pass through the outer membrane proteins of Gram negative bacteria. If the integrity of the membrane of a gram negative bacterium is not good this bacterium will become sensitive to vancomycine.
Hope this will help you
Dear Ashraf,
you can test the sensitivity of your bacteria to different antibiotic families. If your bacteria are sensitive to all, this means that their membrane is damaged. We use vancomycin,erythomycin, rifampicin, bacitracine, novobiocin. If we take the example of vancomycin : Gram negative bacteria are normally resistant to vancomycin. This is a large molecule that cannot pass through the outer membrane proteins of Gram negative bacteria. If the integrity of the membrane of a gram negative bacterium is not good this bacterium will become sensitive to vancomycine.
Hope this will help you
you can use SDS as well. Also A simple approcach will be, if you start to grow them in parallel to wild type and the OD values are not rising as compared to the OD and you doubt that the cell death is because of the membrane integrity, u can spin them and in the sup you will see many outer membrane protein. In parallel you can also perform antibiotic sensitivity keeping in mind the size of the antibiotics and iclude sds.
Thanks lot for suggestion, Prof. Isabelle Virlogeux-Payant and Sadeeq Ur Rahman
In addition to above, you can use a fluorescent dye NPN to quantitatively measure OM permeability. http://www.ncbi.nlm.nih.gov/pubmed/10735988
- Badges
- Science topic
- Similar topics
- Engineering
- Chemical Engineering
- Membranes