Team:The Agency Escondido/project

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<li><a href="https://2013hs.igem.org/Team:The_Agency_Escondido">About</a></li>
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    <li>
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<li><a href="https://2013hs.igem.org/Team:The_Agency_Escondido/notebook">Notebook</a></li>
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        <a href="https://2013hs.igem.org/Team:The_Agency_Escondido">About</a>
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<li><a href="https://2013hs.igem.org/Team:The_Agency_Escondido/safety">Safety</a></li>
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        <ul>
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<li> Project </li>
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            <li><a href="https://2013hs.igem.org/Team:The_Agency_Escondido">General</a></li>
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<li><a href="https://2013hs.igem.org/Team:The_Agency_Escondido/sandiego">San Diego</a></li>
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            <li><a href="https://2013hs.igem.org/Team:The_Agency_Escondido/sandiego">San Diego</a></li>
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<li><a href="https://2013hs.igem.org/Team:The_Agency_Escondido/ourteam">Our Team</a></li>
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            <li><a href="https://2013hs.igem.org/Team:The_Agency_Escondido/ourteam">Our Team</a></li>
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        <a href="https://2013hs.igem.org/Team:The_Agency_Escondido/project"><u>Project</u></a>
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        <ul>
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            <li><a href="https://2013hs.igem.org/Team:The_Agency_Escondido/project/rock">Rock</a></li>
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            <li><a href="https://2013hs.igem.org/Team:The_Agency_Escondido/project/paper">Paper</a></li>
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            <li><a href="https://2013hs.igem.org/Team:The_Agency_Escondido/project/scissors">Scissors</a></li>
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        </ul>
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        <a href="https://2013hs.igem.org/Team:The_Agency_Escondido/notebook">Notebook</a>
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        <ul>
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            <li></li>
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        </ul>
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    <li>
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        <a href="https://2013hs.igem.org/Team:The_Agency_Escondido/safety">Safety</a>
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        <ul>
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            <li></li>
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<h2>
<h2>
Project Description
Project Description
</h2>
</h2>
<p>
<p>
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Our project seeks to create two identical triads of transparent bacteria strains with complementary hydrophobic inducer production to emulate the game of "rock, paper, scissors" (Hydrophobic proteins can move through plasma membranes). Each triad will correspond to a group: group A and group B. Each type of bacteria will produce a specific protein that will act as an inducer to one and only one other bacteria strain(rock strain, paper strain, and scissors strain). Each triad will have different reporter genes designating its champion color. When two bacteria strains are combined in an agar plate, each protein secreted by the victorious strain, when acting as an inducer, changes the color of the cells on the plate identifying the champion.
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Our project seeks to create two identical triads of transparent bacteria strains with complementary hydrophobic inducer production to emulate the game of "rock, paper, scissors" (Hydrophobic proteins can move through plasma membranes). Each triad will correspond to a group: group A and group B. Each type of bacteria will produce a specific protein that will act as an inducer to one and only one other bacteria strain(rock strain, paper strain, and scissors strain). Each triad will have different reporter genes designating its champion color. When two bacteria strains are combined in an agar plate, each protein secreted by the victorious strain, when acting as an inducer, changes the color of the cells on the plate identifying the champion. Through this game, we will learn of the inducers' solubility in water. Only hydrophobic inducers can permeate the cell and induce an operon in a foreign cell. We developed our <a href="http://casp.agency-stem.org/media/G13-LP0013-40.pdf">paper protocol</a>, <a href="http://casp.agency-stem.org/media/G13-LP0009-21.pdf">rock protocol</a>, and <a href="http://casp.agency-stem.org/media/G13-LP0010-57.pdf">scissors protocol</a> before we began our project.
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<p>At the time of writing we are about 75% done with our project. We still need to complete the final assembly of the "scissors" strain and need to complete the second and third assembly of the "rock" and "paper" strains. We anticipate completion by the end of the summer.</p>
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<a id="rock"></a>
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    <th><h2>Contents</h2></th>
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<area shape ="rect" coords ="466,0,700,650" href ="https://2013hs.igem.org/Team:The_Agency_Escondido/project/scissors" alt="Scissors" />
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      <td><a href="#rock"> Rock </a> </td>
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      <td><a href="#paper"> Paper </a> </td>
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      <td><a href="#scissors" > Scissors </a> </td> 
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<a id="rock"></a>
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<h2> Rock </h2>
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<p>The "rock" bacteria strain will have a CFP (<a href = "http://partsregistry.org/Part:BBa_E0020">part E0020</a>) and promoter (<a href = "http://partsregistry.org/Part:BBa_R0080">part R0080</a>). It will contain the LuxR gene (<a href = "http://partsregistry.org/Part:BBa_C0062">part C0062</a>) to induce the "paper" strain.</p>
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<p>Andrew Buss completed the cell competency protocol from the previous
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day, spinning down 8 mL total in sequence.
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With the arrival of ampicillin, Andrew Buss streaked BBa_S01022 onto a
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quadrant of an ampicillin agar plate.
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Andrew Buss began transforming DNA for later assemblies into cells,
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using 5 uL of resuspended DNA per sample in water as suggested by the
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Parts Registry protocol. If the transformation failed with 5 uL, then 40
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uL containing 1 ug could be used in another protocol. 50 pg of DNA from
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the transformation efficiency kit was used as an RFP control. The second
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and last centrifugation in the protocol was carried out using 0.6 mL
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tubes. Unfortunately, the rotor in the centrifuge was designed for 1.5
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mL tubes. When centrifuging, three of the tubes were forced open, and
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fell through the holes in the rotor. These three samples were lost.
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Andrew Buss restarted transformation, using the same protocol. Cells
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were left to incubate overnight in LB containing the appropriate
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antibiotics: the transformation control and pSB1C3 used chloramphenicol,
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while the remainder</p>
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<a id="paper"></a>
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<h2> Paper </h2>
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<p>The "paper" bacteria strain will have an GFP (<a href = "http://partsregistry.org/Part:BBa_E0040">part E0040</a>) and promoter (<a href = "http://partsregistry.org/Part:BBa_I1051">part I1051</a>). It will contain the Lambda cl repressor gene (<a href = "http://partsregistry.org/Part:BBa_C0051">part C0051</a>) to induce the "scissors" strain.</p>
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<a id="scissors"></a>
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<h2> Scissors </h2>
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<p>The "scissors" bacteria strain will have an RFP (<a href = "http://partsregistry.org/Part:BBa_E1010">part E1010</a>)and promoter (<a href = "http://partsregistry.org/Part:BBa_I12006">part I12006</a>). It will contain the AraC gene (<a href = "http://partsregistry.org/Part:BBa_C0080">part C0080</a>) to induce the "rock" strain.</p>
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Latest revision as of 04:15, 22 June 2013