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− | <div style = 'padding-left: | + | <div style = 'padding-left: 14%; padding-bottom: 10px;font-size: 25px' ><b>Overview </b></div> |
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<div style = 'padding-right: 14%; padding-left: 14%; text-indent: 50px;line-height: 25px;' >Recall that unlike steady-state concentration, in our simple kinetic model a given gene's expression speed is defined a function of degradation rate alone. | <div style = 'padding-right: 14%; padding-left: 14%; text-indent: 50px;line-height: 25px;' >Recall that unlike steady-state concentration, in our simple kinetic model a given gene's expression speed is defined a function of degradation rate alone. | ||
− | This implies that it should be possible to readjust our steady-state value back up to its original expression level by increasing protein production rate, without affecting the associated speed change. After determining that we could successfully increase gene expression speed using our characterization parts, we next decided to test whether it was actually possible to adjust the steady-state fluorescence of one of our characterization constructs back to its previous (no degradation) condition. While we anticipate that a real-world implementation of a readjustment to steady-state would probably be implemented through a change in promoter or RBS strength, we decided to our protein production parameter by using a different concentration of inducer. This is analogous to either an RBS or promoter swap because we model protein production as an aggregate of transcription and translation | + | This implies that it should be possible to readjust our steady-state value back up to its original expression level by increasing protein production rate, without affecting the associated speed change. After determining that we could successfully increase gene expression speed using our characterization parts, we next decided to test whether it was actually possible to adjust the steady-state fluorescence of one of our characterization constructs back to its previous (no degradation) condition. While we anticipate that a real-world implementation of a readjustment to steady-state would probably be implemented through a change in promoter or RBS strength, we decided to our protein production parameter by using a different concentration of inducer. This is analogous to either an RBS or promoter swap because we model protein production as an aggregate of transcription and translation. |
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− | <div style = 'padding-left: | + | <div style = 'padding-left: 14%; padding-bottom: 10px;font-size: 25px' ><b>Results</b></div> |
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− | <div style = 'padding-right: | + | <div style = 'padding-right: 14%; padding-left: 14%; text-indent: 50px;line-height: 25px;' >While we previously showed that we could increase the gene expression speed of our inducible mScarlet-I constructs (Figure 1A), it is important to note that the degradation needed to create this speed increase also causes a reduction in steady state protein concentration (Figure 1B).</div> |
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− | <div style = 'padding-right: | + | <div style = 'padding-right: 14%; padding-left: 14%; text-indent: 50px;line-height: 25px;' >To show that it is in fact possible to maintain our degradation mediated increase in gene expression speed while also maintaining existing steady state protein concentration, we determined the protein production parameter (ATC concentration) required to return our ATC inducible pTet mScarlet pdt E characterization construct to its previous steady state protein concentration (Figure 2A). After restoring the steady state protein concentration to it's no-degradation control, we still see the same increase in gene expression speed (Figure 2B). This demonstrates that we can tune gene expression speed while maintaining desired steady-state concentrations of our proteins. </div> |
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Latest revision as of 03:39, 2 November 2017