Difference between revisions of "Team:UFSCar-Brasil/Modeling"

 
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            Modelling
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        Modeling
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          <h2>What we do</h2>
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      <h2 class="ui teal header">Mathematics predicting and explaining events towards repellent</h2>
          <a href="#overview" class="ui huge inverted primary basic button">Overview<i class="right arrow icon"></i></a>
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  <h3 class="ui header" id="overview">Overview</h3>
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  <p> Modeling was made using differential calculus with analytical geometry approaches. Tridimensional solutions were a resource used to explain the multivariable systems obtained in our analysis. This 'joint approach' was chosen to comprise all expected data
          <h3 class="ui header" id="overview">Overview</h3>
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    and provide useful evidence of non-validated data. First of all, Bug Shoo is a product and we must answer several questions not testable in lab before manufacturing tracking, such as the product expiration date. Besides this,
         
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    some project parts have a complex functioning as kill switch and a mathematical model could help to determine the effective time of product working. The effectiveness of the product is another important question, for this, some economic solutions,
 +
    e.g. reduction of PEG 6000 in final formula, optimal zinc chloride concentrations and another parameters could be adjusted with modeling aid. Main used softwares were OriginLab and MatLab. In this sense, we still want to determine the current project
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    as predictive since some parts of it were not produced and the modeling still is passive to work.</p>
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  <h3 class="ui header" id="parts">Plasmolysis</h3>
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  <p>Main purpose of plasmolysis modeling was to determine a relative point, where we have the longest time of bacterial viability and lowest possible percentage of PEG 6000. In order to do that, it was carried out a plasmolysis experiment in our microbiology
 +
    laboratory for some weeks, along we collected the data. Collected data were processed and analyzed to find a best-fit model. Using it, specific values were simulated to generate a mathematical surface. Finally, tangent lines to surface revealed the
 +
    optimal point. Manufacturing tracking is known by its practical nature, and this study was significant to determine the conditions of product storage, besides revealing a technique very useful to other teams in future. Furthermore, this approach provided
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    a prediction of optimal features for maximal product expiration date.
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       <a href="https://2015.igem.org/wiki/index.php?title=Team:UFSCar-Brasil/part1.html" class="ui teal big button">See more! <i class="right arrow icon"></i></a>
 
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          <h3 class="ui header" id="parts">Part I</h3>
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          <p>The purpose of this part of the modeling was to determine what would be the optimum absorbance and with it we would have funded the point relative to the longest time and the lowest percentage of PEG possible. For that were done some weeks of the experiment in the micro-biology laboratory. Then the data were processed and analyzed in order to find which one or ones would be that points. The analysis consisted initially in a model for which the system is most suited. After that, were found specific values which we analyze the simulated surface. Finally, we find the lines that tangents the surface where the point could be found. This study was of great significance when we realize that our project aims to find in stores and with this analysis is made possible to predict the validity of the product.
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  <h3 class="ui header" id="result">Protein Solubilization Toolkit</h3>
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  <p>In this section, we have developed a statistical approach to study the chaperone efficiency in protein folding, in our specific case, limonene synthase folding. This is extensive to all projects with systems of crossed elements effects. As we have several
          <h3 class="ui header" id="result">Part II</h3>
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    chaperones acting in the same final product, the measurement of each effect is complex and could be ineffective. In this sense, we have developed a best-fit model of chaperone arrangement taking in account the folded protein yield. To this, Euclidian
          <p>Desenvolvemos uma abordagem estatística para o estudo da eficiência das chaperonas no enovelamento da limoneno sintase. Com o intuito de criar uma método que melhor descreva a combinação de chaperonas com base nos dados de rendimento, criamos um dendograma que ilustra o agrupamento delas com base na distancia estatística dos seus rendimentos. Depois foi resolvido um sistema linear com todas as relações e encontrou-se constantes estatísticas de cada componente.</p>
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    mathematical distances were used to clustering analysis generating a dendogram which was used to determine the main groups and statistical differences. After this, the system was solved with a linear system where all relations were input to generate
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    an output of mathematical effect constants for each component.</p>
  
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       <a href="https://2015.igem.org/wiki/index.php?title=Team:UFSCar-Brasil/part2.html" class="ui teal big button">See more! <i class="right arrow icon"></i></a>
 
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          <h3 class="ui header" id="judging">Part III</h3>
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          <p>One of our prime objective is to describe the activity of uspA promoter ( Universal Stress Protein A promoter) when exposed to osmotic chock compared with J23101 promoter. To quantify more precisely this behavior we adjust experimental points with general exponential functions and also related PEG concentration’s date with osmotic pressure. With the proper fitted curves, we modeled the concentration’s fall of Zn 2+ from external environment by import the metal to intracellular environment and gradually build up the smtA protein aiming estimate the approximate time for begin the death cell process, that initiate with the release of our killswitch’s promoter region due to the absence of Zn in cellular environmental to maintain the zur factor repressing ufscarA promoter. With the unblocked promoter, the transcription of death genes starts, metabolism and cell integrity is compromised leading to cell death.</p>
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<h3 class="ui center aligned header">Our amazing sponsors!</h3>
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  <h3 class="ui header" id="judging">Kill Switch</h3>
<img class="ui centered massive image" src="https://static.igem.org/mediawiki/2015/archive/0/0e/20150908223157!UFSCar-Brasil_logos.png">
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   <p>One of our prime objectives was to describe the uspA promoter activity (<a href="http://parts.igem.org/Part:BBa_K1620000">BBa_K1620000</a> and <a href="http://parts.igem.org/Part:BBa_K1620005">BBa_K1620005</a>) when exposed to osmotic chock compared
 
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    with a constitutive promoter, J23101 derived construct (<a href="http://parts.igem.org/Part:BBa_K1620006">BBa_K1620006</a>). First of all, the adjust of fluorescence points to exponential functions were done to give fluorescence values / GFP molecule
 +
    quantities related to PEG concentrations or even calculated osmotic pressures. Proper fitted curves were used to model concentration of Zn(II) in culture medium along time (describing a decay curve). The final concentration of Zn(II) in culture medium
 +
    will be lower due to metal ions import to intracellular environment followed by immobilization through gradual association to smtA protein (<a href="http://parts.igem.org/Part:BBa_K1620007">BBa_K1620007</a>). This section aims estimate the approximate
 +
    time for begin the death cell process, well known when Zn(II) concentration tends to minimal and the Zasp element (<a href="http://parts.igem.org/Part:BBa_K1620001">BBa_K1620001</a>) starts promotion of killer genes. This period is marked by releasing
 +
    of our killswitch’s promoter region due to absence of intracellular Zn(II) available to maintain zur factor (<a href="http://parts.igem.org/Part:BBa_K1620004">BBa_K1620004</a>) repressing Zasp element (<a href="http://parts.igem.org/Part:BBa_K1620001">BBa_K1620001</a>).
 +
    This section still reveals to us how much time the product will work after its activation.</p>
  
 
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Latest revision as of 20:05, 18 September 2015

Modeling

Mathematics predicting and explaining events towards repellent

Overview

Modeling was made using differential calculus with analytical geometry approaches. Tridimensional solutions were a resource used to explain the multivariable systems obtained in our analysis. This 'joint approach' was chosen to comprise all expected data and provide useful evidence of non-validated data. First of all, Bug Shoo is a product and we must answer several questions not testable in lab before manufacturing tracking, such as the product expiration date. Besides this, some project parts have a complex functioning as kill switch and a mathematical model could help to determine the effective time of product working. The effectiveness of the product is another important question, for this, some economic solutions, e.g. reduction of PEG 6000 in final formula, optimal zinc chloride concentrations and another parameters could be adjusted with modeling aid. Main used softwares were OriginLab and MatLab. In this sense, we still want to determine the current project as predictive since some parts of it were not produced and the modeling still is passive to work.

Plasmolysis

Main purpose of plasmolysis modeling was to determine a relative point, where we have the longest time of bacterial viability and lowest possible percentage of PEG 6000. In order to do that, it was carried out a plasmolysis experiment in our microbiology laboratory for some weeks, along we collected the data. Collected data were processed and analyzed to find a best-fit model. Using it, specific values were simulated to generate a mathematical surface. Finally, tangent lines to surface revealed the optimal point. Manufacturing tracking is known by its practical nature, and this study was significant to determine the conditions of product storage, besides revealing a technique very useful to other teams in future. Furthermore, this approach provided a prediction of optimal features for maximal product expiration date.

Protein Solubilization Toolkit

In this section, we have developed a statistical approach to study the chaperone efficiency in protein folding, in our specific case, limonene synthase folding. This is extensive to all projects with systems of crossed elements effects. As we have several chaperones acting in the same final product, the measurement of each effect is complex and could be ineffective. In this sense, we have developed a best-fit model of chaperone arrangement taking in account the folded protein yield. To this, Euclidian mathematical distances were used to clustering analysis generating a dendogram which was used to determine the main groups and statistical differences. After this, the system was solved with a linear system where all relations were input to generate an output of mathematical effect constants for each component.

Kill Switch

One of our prime objectives was to describe the uspA promoter activity (BBa_K1620000 and BBa_K1620005) when exposed to osmotic chock compared with a constitutive promoter, J23101 derived construct (BBa_K1620006). First of all, the adjust of fluorescence points to exponential functions were done to give fluorescence values / GFP molecule quantities related to PEG concentrations or even calculated osmotic pressures. Proper fitted curves were used to model concentration of Zn(II) in culture medium along time (describing a decay curve). The final concentration of Zn(II) in culture medium will be lower due to metal ions import to intracellular environment followed by immobilization through gradual association to smtA protein (BBa_K1620007). This section aims estimate the approximate time for begin the death cell process, well known when Zn(II) concentration tends to minimal and the Zasp element (BBa_K1620001) starts promotion of killer genes. This period is marked by releasing of our killswitch’s promoter region due to absence of intracellular Zn(II) available to maintain zur factor (BBa_K1620004) repressing Zasp element (BBa_K1620001). This section still reveals to us how much time the product will work after its activation.

Our amazing sponsors!