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Team:Calgary/21 August 2009
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Beyond that I also sat around and played with Excel to see what kind of graphs I could generate with my data and what use it would be. Data was re-arrayed with a macro donated by Margot from the Surette lab (''MANY'' thanks with everything you have helped us with!!). | Beyond that I also sat around and played with Excel to see what kind of graphs I could generate with my data and what use it would be. Data was re-arrayed with a macro donated by Margot from the Surette lab (''MANY'' thanks with everything you have helped us with!!). | ||
| - | [[Image:Calgary_AI2-(1).jpg | | + | [[Image:Calgary_AI2-(1).jpg | 600px]] |
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'Figure 1.'Luminescence readings from an autoinducer bioassay using cell-free supernatants from overnights of various bacterial strains with Vibrio harveyi MM32 as the reporter strain. MM32 is responsive (i.e. luminescent) to the AI-2 present in the supernatant of V. harveyi BB120, a wildtype strain, and BB152 (BB120 luxLM::Tn5)2, a strain deficient in AHL production. Salmonella typhimurium 14028 produces AI-2 but due to differences in boration1, specificity to the V. harveyi reporter strain, and therefore luminescence, decreases. S. typhimurium SS007 (14028 luxS::T-POP)3 is incapable of producing AI-2 and serves as a negative control. E. coli DH5α has been reported as lacking the ability to produce AI-24. The lack of a peak in luminescence for MM32 grown with DH5α supernatant validates using DH5α as the chassis for our system. Our signalling system in DH5α will be responsive solely to exogenously added AI-2 derived from BB152 as it only produces AI-2. Cell-free supernatants were made by centrifugation of overnight cultures and filtering it with a 0.2μm (indicated by an ‘F’). Controls indicate the importance of filtering the supernatant (data not shown). The graph indicates an average obtained from 3 trials. | 'Figure 1.'Luminescence readings from an autoinducer bioassay using cell-free supernatants from overnights of various bacterial strains with Vibrio harveyi MM32 as the reporter strain. MM32 is responsive (i.e. luminescent) to the AI-2 present in the supernatant of V. harveyi BB120, a wildtype strain, and BB152 (BB120 luxLM::Tn5)2, a strain deficient in AHL production. Salmonella typhimurium 14028 produces AI-2 but due to differences in boration1, specificity to the V. harveyi reporter strain, and therefore luminescence, decreases. S. typhimurium SS007 (14028 luxS::T-POP)3 is incapable of producing AI-2 and serves as a negative control. E. coli DH5α has been reported as lacking the ability to produce AI-24. The lack of a peak in luminescence for MM32 grown with DH5α supernatant validates using DH5α as the chassis for our system. Our signalling system in DH5α will be responsive solely to exogenously added AI-2 derived from BB152 as it only produces AI-2. Cell-free supernatants were made by centrifugation of overnight cultures and filtering it with a 0.2μm (indicated by an ‘F’). Controls indicate the importance of filtering the supernatant (data not shown). The graph indicates an average obtained from 3 trials. | ||
Revision as of 20:22, 18 September 2009
UNIVERSITY OF CALGARY
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