Team:LCG-UNAM-Mexico/Notebook/2008-September

From 2008.igem.org

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  <strong>General checkup </strong><br />
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<br />
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<em>Simulation:</em> The previous crisis was overcome... We are walking fine, however, there are  still some parameters missing, but it has been decided that they will be  estimated by adjusting the model and experimentally when we have the  opportunity.<br />
 +
<br />
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<em>Data:</em> There are a few  missing parameters and we know that most of them  are not available, so we have given up the search... What has yet to be defined in  terms of data are concentrations of AiiA and LuxR, considering that  they are preserved constants in the model.</p>
 +
<p>We will also design  experiments to obtain the missing parameters through the experimental measurements (viable). <br />
 +
  <br />
 +
<em>Analysis:</em> Stechiometric matrix: There was a meeting today in the morning with Osbaldo to review its analysis, we will share the information as soon as possible. </p>
 +
<p> <em>Sensitivity  analysis:</em> Although we don't understand yet the particular units in which  SimBiology returns the results, the first graphics that show the  basic parameters of the model are ready and they are the ones involved in the  degradation of AHL and dimerization of it with LuxR ( the start of the cascade) and the ones regarding the entry and exit of Nickel. We must do this  analysis recurrently, as we have seen that this is sensitive to the  initial concentrations of metabolites, which are not yet fully defined. </p>
 +
<p><em>States and stationary Jacobian</em>: They were stopped briefly because  we need the parameters for further analysis.
 +
    <br />
 +
  <br />
 +
  <strong>Experimental </strong><br />
 +
  <br />
 +
  <em>Requirements:</em> Urgent! We need to send the oligos required to  synthesis; We are working on it, but  let's consider this a priority. <br />
 +
  <br />
 +
  <em>Electrodes:</em> The device is not ready.<br />
 +
  <br />
 +
  <em>Design of experiments</em>: The first meeting will be today. <br />
 +
</p>
 +
<p><em>Funds &amp; Jambouree:</em> We are still waiting for some sponsors to reply.<br />
 +
  <br /></p>
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Revision as of 22:48, 7 October 2008

LCG-UNAM-Mexico:Notebook/September

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September

2008-09-3


Lac promoter synthesis rate

van Hoek F, Hogeweg P (2007) The effect of stochasticity on the Lac Operon: An evolutionary perspective. PLoS Comput Biol 3 (6): E111

The article's objective, as you can infer from the title, is to evaluate the effect of stochasticity in the evolution of a Promoter. In order to do so they built a comprehensive model including every parameter involved in Transcription and translation. They measure some parameters but they depend mostly on literature to define them.

They both do a deterministic and a Stochastic analysis. To generate a Stochastic model they added one parameter, the average burst size of protein translation (protein translation occurs in bursts, after a several mRNA is synthesized proteins can be translated from the same mRNA). This was possible because when an mRNA molecule is translated it can not be degraded. Therefore after each translation it can either be translated again (p) or be degraded (1-p). This suggests that protein production occurs in bursts with a burst size geometrically distributed. After they compared their noise levels in the model with the experimental noise measurements they found good agreement.

Transcription they used to model a two-dimensional Hill-function dependent on the cAMP concentration and alloctase. (repressa the glucose and lactose Operon via cAMP activates the Operon via allolactose).

They use 11 biochemical parameters, including three of special importance for us:

a, when the rate Transcription RNA Polymerase is bound to the DNA, but CRP and Laci are not. evolva: initial value: 1.1 × 10-7 mM / min

b, The Transcription rate when both RNA Polymerase and CRP are bound, but Laci is not bound to the DNA. evolva: initial value: 2.2 × 10-5 mM / min

c,''leakiness,''the Transcription rate when RNA Polymerase is not bound to the DNA. evolva: initial value 5.5 × 10-10 mM / min

They modele binomially protein degradation, assuming that cell when to divide proteins are divided randomly between the cells. However in a population of non-Dividing cells this "dilution" can not be taken in account.

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2008-09-23



General checkup

Simulation: The previous crisis was overcome... We are walking fine, however, there are still some parameters missing, but it has been decided that they will be estimated by adjusting the model and experimentally when we have the opportunity.

Data: There are a few missing parameters and we know that most of them are not available, so we have given up the search... What has yet to be defined in terms of data are concentrations of AiiA and LuxR, considering that they are preserved constants in the model.

We will also design experiments to obtain the missing parameters through the experimental measurements (viable).

Analysis: Stechiometric matrix: There was a meeting today in the morning with Osbaldo to review its analysis, we will share the information as soon as possible.

Sensitivity analysis: Although we don't understand yet the particular units in which SimBiology returns the results, the first graphics that show the basic parameters of the model are ready and they are the ones involved in the degradation of AHL and dimerization of it with LuxR ( the start of the cascade) and the ones regarding the entry and exit of Nickel. We must do this analysis recurrently, as we have seen that this is sensitive to the initial concentrations of metabolites, which are not yet fully defined.

States and stationary Jacobian: They were stopped briefly because we need the parameters for further analysis.  

Experimental

Requirements: Urgent! We need to send the oligos required to synthesis; We are working on it, but let's consider this a priority.

Electrodes: The device is not ready.

Design of experiments: The first meeting will be today.

Funds & Jambouree: We are still waiting for some sponsors to reply.