Difference between revisions of "Team:Vilnius-Lithuania/Results"
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<p class="text-justify">For the fluorescence measurement experiments we took advantage of protocol that was provided by the iGEM headquarters for the InterLab Measurement experiment. We transformed JM109 bacteria with our new constructs that were cloned into two separate compatible pasmid vectors. In theory, our regulatory unit upon induction with IPTG should produce cI repressor, which, in turn, should downregulate the expression of the GFP gene (under the control of cI regulated promoter). We seeked to quantify this by growing cell cultures overnight with different IPTG concentrations. We also used a negative and a positive control. </p> | <p class="text-justify">For the fluorescence measurement experiments we took advantage of protocol that was provided by the iGEM headquarters for the InterLab Measurement experiment. We transformed JM109 bacteria with our new constructs that were cloned into two separate compatible pasmid vectors. In theory, our regulatory unit upon induction with IPTG should produce cI repressor, which, in turn, should downregulate the expression of the GFP gene (under the control of cI regulated promoter). We seeked to quantify this by growing cell cultures overnight with different IPTG concentrations. We also used a negative and a positive control. </p> | ||
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+ | <p class="text-justify">We determined that this device is sensitive to cI. It can be seen that upon induction of cI expression, there is a significant drop in relative fluorescence, the general tendency is that fluorescence signal depletes with increasing concentrations of IPTG, however, we see great variation in our data. Similar data was obtained during replications of this experiment. We suggest that BBa_K195613 is indeed sensitive to cI and the effects of cI on transcription levels from this promoter are somewhat titratable, however, we claim that this system (BBa_K077039 + BBa_K195613) is highly unpredictable. We suggest that this unpredictability might be due to the low translation levels of cI protein (weak RBS) combined with its instability (LVA tail) and its nature of a dominant repressor. These effects combined might lead to an effect in which changes in a small population of repressor might have great effects on the whole system. </p> | ||
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Revision as of 06:13, 18 September 2015