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<div style = 'padding-right: 190px; padding-left: 190px; text-indent: 50px;line-height: 25px;' >Figure 1 shows the raw fluorescence (MECY) of our ATC inducible mScarlet-I pdt constructs. Stronger pdts (earlier letters) show decreased fluorescence due to degradation. As a proof of concept, we show that our inducible pdt F construct can have it's raw fluorescence readjusted to previous values without any loss of gene expression speed (Figure 2)</div>
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<div style = 'padding-right: 190px; padding-left: 190px; text-indent: 50px;line-height: 25px;' >Temp. Figures go to left (for #1, and below for #2). Link 1 https://2017.igem.org/File:T--William_and_Mary--mScarlet-I_Speed_MECY_Full.png Figure 1: Raw fluorescence values of mScarlet-I pdt constructs. Each data point represents the geometric mean of three biological replicates, and the shaded region represents one geometric standard deviation above and below the mean Figure 2: Raw fluorescence (A) and normalized fluorescence (B) for the original (50ng/mL) vs adjusted (85ng/mL) strength mScarlet-I pdt 3E construct. Each data point represents the geometric mean of three biological replicates, and the shaded region represents one geometric standard deviation above and below the mean </div>
Revision as of 03:23, 31 October 2017
Overview
While we have demonstrated that we can see a real speed change, you might remember that in our math section we noted that the steady state value is given as the production rate divided by the degradation rate. This means that though we are increasing the speed of gene expression, we are also decreasing the steady state value. While some applications may only care about a gene’s expression an on or off signal, and not about the magnitude of expression, we wanted our system to be usable in any system. Since in our model gene expression speed is only regulated by degradation, it should be possible to readjust our steady state value back up to its original expression. Using pdt E as an example, we measured the time to steady state with and without mf-Lon at a given ATC induction level. We then showed that by increasing the ATC concentration (increasing production rate), we can return the steady state of the protease condition to the no protease condition while maintaining the same speed change, exactly as our model predicts.
Modeling?
Results
Figure 1 shows the raw fluorescence (MECY) of our ATC inducible mScarlet-I pdt constructs. Stronger pdts (earlier letters) show decreased fluorescence due to degradation. As a proof of concept, we show that our inducible pdt F construct can have it's raw fluorescence readjusted to previous values without any loss of gene expression speed (Figure 2)
Temp. Figures go to left (for #1, and below for #2). Link 1 https://2017.igem.org/File:T--William_and_Mary--mScarlet-I_Speed_MECY_Full.png Figure 1: Raw fluorescence values of mScarlet-I pdt constructs. Each data point represents the geometric mean of three biological replicates, and the shaded region represents one geometric standard deviation above and below the mean Figure 2: Raw fluorescence (A) and normalized fluorescence (B) for the original (50ng/mL) vs adjusted (85ng/mL) strength mScarlet-I pdt 3E construct. Each data point represents the geometric mean of three biological replicates, and the shaded region represents one geometric standard deviation above and below the mean
