Difference between revisions of "Part:BBa K554001:Experience"
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<h4><b><span>BBa_K554001 Testing</span></b></h4> | <h4><b><span>BBa_K554001 Testing</span></b></h4> | ||
| − | < | + | <p>The flhDC promoter <a href="https://parts.igem.org/Part:BBa_K554001" target="_blank" alt="BBa_K554001">BBa_K554001</a> from the 2011 UNICAMP-EMSE Brazil iGEM team was what we needed. |
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In order to test the promoter, we ligated the GFP reporter part, <a href="https://parts.igem.org/Part:BBa_I13504" target="_blank">BBa_I13504</a>, downstream of the promoter, transformed it in DH5-α and measured the fluorescence on a plate reader after being induced with varying concentrations of epinephrine at OD=0.1. | In order to test the promoter, we ligated the GFP reporter part, <a href="https://parts.igem.org/Part:BBa_I13504" target="_blank">BBa_I13504</a>, downstream of the promoter, transformed it in DH5-α and measured the fluorescence on a plate reader after being induced with varying concentrations of epinephrine at OD=0.1. | ||
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Fig. 2 shows that at after around OD = 1, fluorescence becomes higher in the cultures containing epinephrine compared to the culture with no epinephrine. On another note, no large increase in fluorescence was seen from increasing the concentration of epinephrine beyond 5 μM, | Fig. 2 shows that at after around OD = 1, fluorescence becomes higher in the cultures containing epinephrine compared to the culture with no epinephrine. On another note, no large increase in fluorescence was seen from increasing the concentration of epinephrine beyond 5 μM, | ||
showing that 5 μM epinephrine is enough to induce the promoter to maximum strength. | showing that 5 μM epinephrine is enough to induce the promoter to maximum strength. | ||
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Revision as of 16:39, 19 September 2015
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Applications of BBa_K554001
BBa_K554001 Testing
The flhDC promoter BBa_K554001 from the 2011 UNICAMP-EMSE Brazil iGEM team was what we needed.
In order to test the promoter, we ligated the GFP reporter part, BBa_I13504, downstream of the promoter, transformed it in DH5-α and measured the fluorescence on a plate reader after being induced with varying concentrations of epinephrine at OD=0.1.
Figure 2. Fluorescence of DH5-α cultures when flhDC promoter was induced at varying concentrations of (-)-epinephrine.
Fig. 2 shows that at after around OD = 1, fluorescence becomes higher in the cultures containing epinephrine compared to the culture with no epinephrine. On another note, no large increase in fluorescence was seen from increasing the concentration of epinephrine beyond 5 μM, showing that 5 μM epinephrine is enough to induce the promoter to maximum strength.
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