Team:ETH Zurich/modeling/int

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Replacing every occurence of Bxb1 by ΦC31 gives the set of parameters for ΦC31. The same status can be applied to those parameters.  
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Substituting ΦC31 for Bxb1 gives the set of parameters for ΦC31. The same status can be applied to those parameters.  
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Even if degradation rates were not determined specifically for the serine integrases and their dimerized form, degradation rates of proteins in ''E. coli'' are available. We assume that the degradation rates of dimerized forms are two times higher than the degradation rates of monomers. Typically, d<sub>DBxb1</sub> = 2*d<sub>Bxb1</sub> To characterize integrase behavior, we focus on finding the parameters for dimerization and DNA-binding.
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Even if degradation rates were not determined specifically for the serine integrases and their dimerized form, degradation rates of proteins in ''E. coli'' are available. We assume that the degradation rates of dimerized forms are two times higher than the degradation rates of monomers. Typically, d<sub>DBxb1</sub> = 2*d<sub>Bxb1</sub>. To characterize integrases behavior, we focused on estimating the parameters for dimerization and DNA-binding.
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=== Data ===
=== Data ===
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The parameter fitting is based on data from Bonnet's paper<sup>[[Team:ETH_Zurich/project/references|[9]]]</sup>. Their experimental setup is different from ours. However, they experimentally retrieve a transfer function between aTc and Bxb1 switching rate. Here is the figure of interest in our case.
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The parameter fitting is based on the data available from Bonnet's paper<sup>[[Team:ETH_Zurich/project/references|[9]]]</sup>. Their experimental setup is different from ours. They induce the production of the integrase Bxb1 with aTc. Bxb1 can then flip a terminator put between a constitutive promoter and the gene for a reporter protein. They experimentally retrieve a transfer function between aTc and Bxb1 switching rate. Here is the figure of interest in our case.
[[File:ETH Zurich Bonnet S4.jpg|center|400px|thumb|'''Figure 2''' Transfer function from aTc to Bxb1. The experimental data corresponds to the points. They fitted this data with their own model. Supplementary figure S4 of Bonnet's paper ''Amplifying Genetic Logic Gates''<sup>[[Team:ETH_Zurich/project/references|[9]]]</sup>.]]
[[File:ETH Zurich Bonnet S4.jpg|center|400px|thumb|'''Figure 2''' Transfer function from aTc to Bxb1. The experimental data corresponds to the points. They fitted this data with their own model. Supplementary figure S4 of Bonnet's paper ''Amplifying Genetic Logic Gates''<sup>[[Team:ETH_Zurich/project/references|[9]]]</sup>.]]
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=== Assumptions ===
=== Assumptions ===
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;'''Assumption A'''
 
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:The back-reaction from DBxb1 binding to site inactive is considered to be negligible, compared to the flipping rate. That is to say that once a site is active, it can only be flipped. Thus, an active site would only be a transitional state in our whole cell model. As the flipping is not modeled in our integrase subsystem, active sites are not transformed into flipped sites at the end of the information pipeline. Thus, we consider that we can express the switching rate given active site concentration.
 
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;'''Assumption B'''
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We keep the following assumptions from the previous characterization: '''assumption A''', '''assumption B''' and '''assumption C'''. A assumption is made:
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:As switching needs two active sites to be effective (for more information on flipping, check the [https://2014.igem.org/Team:ETH_Zurich/modeling/xor XOR gate page]), the switching rate is approximated to: $${(\frac{SA_{Bxb1}}{S_{TOT}})}^2$$ This approximation is understated by statistical considerations.
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;'''Assumption C'''
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:Given the normalization of the paper, basal rate of production of Bxb1 is not taken into account. Thus, we consider that $$A_L = 0$$
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;'''Assumption E'''
;'''Assumption E'''

Latest revision as of 01:09, 18 October 2014

iGEM ETH Zurich 2014