Bacteria within a biofilm are more effective in phenomena that are detrimental to human well being. Based on the work of Dr. Matin and others, it is hypothesized that due to low-shear fluid effects, such as potentially altered macromolecular folding, and documented changes in gene expression, biofilms under low-shear conditions are different from their conventional counterparts. Using novel adaptations to a rotating wall vessel (RWV) bioreactor, devised in the Matin lab, E. coli biofilms will be cultivated in low-shear fluid environments. What unique genes and proteins are expressed at different developmental stages of the low-shear biofilms will be determined, using DNA microarrays, 2-D gel electrophoresis, and mass spectrometry. Mutants unable to form low-shear biofilms or unable to enhance resistance will be isolated. Use of knockouts in selected genes will further explore the role of individual genes in low-shear biofilm formation and antibiotic resistance. What effect RWVs may have on protein-protein interactions will be determined, using two hybrid and TAP-tagging systems, with suitable proteins. Fundamental and enabling information will result in better control of biofilms for preventing infectious disease. ? ? ?
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