Difference between revisions of "Team:UFSCar-Brasil/achievements.html"

 
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            Achievements
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          <h2>What we do</h2>
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      <h2 class="ui teal header">What we do</h2>
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      <img class="ui small image" src="https://static.igem.org/mediawiki/2015/8/83/UFSCar-Brasil_falcons.PNG" alt=""> Overview
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    <p> During laboratory activities realized by UFSCar-Brasil team, was obtained different results for the proposed project, as well as the contribution with new biobricks for registry and new protocols for scientific community use in forward projects. Based
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      on molecular biology, bioinformatics and microbiology techniques, our Team could propose protocols and biobricks, which assist obtaining protein of difficult expression through the UspA promoter, as well as maintaining bacterial viability at room
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      temperature by plasmolysis induced polyethylene glycol. Thus, we believe that with the expansion of synthetic biology to market manufactures, our protocols represent an important tool in enabling bacteria genetically engineered shelf products.</p>
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    <p>Another important point refer to the following project's module, characterized by overexpression of chaperones during cell growth and the utilization of a biosafety method as Killswitch; Developed by UFSCar-Brasil team, these modules can also represent
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      important tools that envisaged the viability and theoretical basis of the project. We believe that, although many structural and funding problems, attach ourselves to the iGEM spirit and we achieved great achievements, and through impressive results
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      in the laboratory, which are described below, contribute to the development of synthetic biology.</p>
  
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          <h3 class="ui header" id="overview">Overview</h3>
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          <p>Diseases transmitted by insect vectors, such as malaria and dengue, affect significantly the Brazilian population. In our city, São Carlos, this year in summer the public health organ featured a big number of cases. Working in this problem,
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            our project consists in the development of an alternative repellent of that currently sold in the market and more or equally effective in preventing mosquitoes bites transmitted diseases. The main compound in current repellents is DEET (N,
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            N-diethyl-m-toluamide), a toxic molecule which at certain concentrations could be lethal, and therefore must have strict control on their use in products. The main characteristic of our repellent is the long duration when compared to other
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            products and the replacement of the compound DEET by D-limonene. Instead, the D-limonene has low toxicity, is highly volatile and present a pleasant smell.</p>
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          <p>Our proposal is building a bacteria carrying out the production of D-limonene via limonene synthase. To enable long term storage at room temperatures, the bacterial cells that make up the repellent will be plasmolyzed, with suspended metabolism,
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            considered a dormant state. The maintenance of this state will be obtained by a solution of polyethyleneglycol (PEG) that will raise the osmotic pressures. Once in contact with the skin, the PEG solution will be diluted by sweat, inducing
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            an osmotic shock in cells. Then the universal stress protein promoter (UspA) will be activated and will induce the expression of limonene synthase. Beyond PEG, other compounds such as glycerol and metal ions in low concentrations will compose
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            our insect repellent cream, in order to sustain the bacteria to the required metabolites building and posterior enzyme activities. In this way, the distribution of our repellent may become feasible, allowing its use.</p>
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          <p>To overcome those problems with limonene synthase folding, we used constitutive promoters for the expression of chaperones from all Escherichia coli available classes (like ClpB, DnaK and IbpA / IbpB). It will reinforce the stockpile of chaperones
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            naturally produced during osmotic shocks (heat-shock proteins) in bacteria. Our goal is to improve protein solubility, indirectly creatinga toolkit for protein solubility enhancement for future iGEM teams. . Besides, reducing the occurrence
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  <h3 class="ui header" id="parts">Parts</h3>
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            of insoluble bodies and improving the production of limonene.</p>
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  <p>Here UFSCar-Brasil presents submitted parts to registry, allied to those used by us. In this sense, we submitted 11 new biobricks, being: 3 intermediate parts, 2 composite parts and 6 new simple parts. Our main goal was to design, build and test
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    <a
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    href="http://parts.igem.org/Part:BBa_K1620000">UspA promoter</a> considered our best part.</p>
  
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          <p>Finally, a way to ensure the biosafety in the use of our repellent and keep a control of bacterial cells is the use of a system of programmed death or Kill Switch. This system will work with two suicide genes: Killer red, a protein which trigger
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            oxidative species generation destroying the DNA (plasmid and chromosomal) allied to a potent RNAse which will avoid any contribution of RNA to host or normal microbiota. These genes will be switch on through an association with a zinc sensitive
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            promoter triggered by zur proteins which associate with zinc and avoid the expression of them allowing the bacterial growing. After long periods of repellent action, bacteria activities will decrease zinc concentration in medium, which allows
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            the gene activation and cells death.</p>
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  <h6 class="ui center aligned header"><b>Figure 1</b>: Genetic circuit of Bug Shoo.</h6>
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          <h3 class="ui header" id="parts">Parts</h3>
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          <p>Diseases transmitted by insect vectors, such as malaria and dengue, affect significantly the Brazilian population. In our city, São Carlos, this year in summer the public health organ featured a big number of cases. Working in this problem,
+
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            our project consists in the development of an alternative repellent of that currently sold in the market and more or equally effective in preventing mosquitoes bites transmitted diseases. The main compound in current repellents is DEET (N,
+
−
            N-diethyl-m-toluamide), a toxic molecule which at certain concentrations could be lethal, and therefore must have strict control on their use in products. The main characteristic of our repellent is the long duration when compared to other
+
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            products and the replacement of the compound DEET by D-limonene. Instead, the D-limonene has low toxicity, is highly volatile and present a pleasant smell.</p>
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    <a href="https://2015.igem.org/wiki/index.php?title=Team:UFSCar-Brasil/parts.html" class="ui big teal button">See more!</a>
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          <h3 class="ui header" id="result">Result</h3>
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  <h3 class="ui header" id="result">Result</h3>
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  <p>Here we presented all results of our work. We obtained new biobricks, generated characterization information. Although do not achieve a functional prototype, we design mathematical methods and lab protocols to further testings. We also report all steps
 +
    of biobricks and testing obtaining. Finally, we also reported plasmolysis experiment, where we could observe a great viability after 7 weeks using a cost effective and simple treatment, which could help transpass in the future our manufacturing tracking
 +
    project into a real product.</p>
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  <div class="center aligned">
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    <a href="https://2015.igem.org/wiki/index.php?title=Team:UFSCar-Brasil/results.html" class="ui big teal button">See more!</a>
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The universal stress protein A is a response of Escherichia coli cells to growth arrest, and its lacking generates cells with defective growth. For more information about this important protein, please feel free to visit the respective Wikigenes page (https://www.wikigenes.org/e/gene/e/948007.html).
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The universal stress protein A promoter (also as known as PuspA) is a well characterized promoter element, inducible under several stress conditions (Prytz et al. 2003; Dyk et al. 1995; Nyström and Neidhardt 1992; 1994). This genetic element is dependent to sigma-70 factor (Nyström and Neidhardt 1992; 1994). Its regulation is done through the concentration of a specific stationary phase allormone, guanosine-5'-diphosphate-3'-diphosphate (ppGp), as described elsewhere (Farewell et al. 1998b). The ppGp activate the transcription of downstream elements through a positive regulation of the β-subunit of RNA polymerase. In this sense, the PuspA element is considered a stationary phase promoter. However, in the work of Prytz et al. (2003) a constitutive transcription promotion was observed.
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  <h3 class="ui header" id="judging">Judging Criteria</h3>
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Diverse conditions make the Escherichia coli cell enter to stress state, like heat shocks, starvation, osmotic stress, ultraviolet light and other conditions. In these situations, the RNA polimerase sigma factors (ropS) trigger the expression of chaperones and other stress protector molecules, in order to help the cell survive. Previous works have showed the power of the element PuspA, like shown in the Table 1.
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  <p>Judging criteria are presented allied to our achievements showing how great our work was!</p>
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          <p>Diseases transmitted by insect vectors, such as malaria and dengue, affect significantly the Brazilian population. In our city, São Carlos, this year in summer the public health organ featured a big number of cases. Working in this problem,
 
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            our project consists in the development of an alternative repellent of that currently sold in the market and more or equally effective in preventing mosquitoes bites transmitted diseases. The main compound in current repellents is DEET (N,
 
−
            N-diethyl-m-toluamide), a toxic molecule which at certain concentrations could be lethal, and therefore must have strict control on their use in products. The main characteristic of our repellent is the long duration when compared to other
 
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            products and the replacement of the compound DEET by D-limonene. Instead, the D-limonene has low toxicity, is highly volatile and present a pleasant smell.</p>
 
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Latest revision as of 21:49, 18 September 2015

Home
Project
Overview
Issue Plasmolysis and Limonene synthase Protein Solubilization Toolkit Kill Switch
Team
Our Team Attributions
Achievements
Overview
Parts Results Judging Criteria
Policy & Practices
Overview
Political Social Educational Safety/Security Meetings Fundraising
Modeling
Overview
Plasmolysis Protein Solubilization Toolkit Kill Switch
Notebook
Protocols Problems Collaborations Safety Acknowledgements

Achievements

What we do

Overview

During laboratory activities realized by UFSCar-Brasil team, was obtained different results for the proposed project, as well as the contribution with new biobricks for registry and new protocols for scientific community use in forward projects. Based on molecular biology, bioinformatics and microbiology techniques, our Team could propose protocols and biobricks, which assist obtaining protein of difficult expression through the UspA promoter, as well as maintaining bacterial viability at room temperature by plasmolysis induced polyethylene glycol. Thus, we believe that with the expansion of synthetic biology to market manufactures, our protocols represent an important tool in enabling bacteria genetically engineered shelf products.

Another important point refer to the following project's module, characterized by overexpression of chaperones during cell growth and the utilization of a biosafety method as Killswitch; Developed by UFSCar-Brasil team, these modules can also represent important tools that envisaged the viability and theoretical basis of the project. We believe that, although many structural and funding problems, attach ourselves to the iGEM spirit and we achieved great achievements, and through impressive results in the laboratory, which are described below, contribute to the development of synthetic biology.

Parts

Here UFSCar-Brasil presents submitted parts to registry, allied to those used by us. In this sense, we submitted 11 new biobricks, being: 3 intermediate parts, 2 composite parts and 6 new simple parts. Our main goal was to design, build and test UspA promoter considered our best part.

Figure 1: Genetic circuit of Bug Shoo.
See more!

Result

Here we presented all results of our work. We obtained new biobricks, generated characterization information. Although do not achieve a functional prototype, we design mathematical methods and lab protocols to further testings. We also report all steps of biobricks and testing obtaining. Finally, we also reported plasmolysis experiment, where we could observe a great viability after 7 weeks using a cost effective and simple treatment, which could help transpass in the future our manufacturing tracking project into a real product.

See more!

Judging Criteria

Judging criteria are presented allied to our achievements showing how great our work was!

See more!

Our amazing sponsors!

Links

iGEM UFSCar

About

Team

Thanks for reading our wiki!

Come Again :)