Team:Chiba/Project/uptake

From 2013.igem.org

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<h2 id="oxidation" style="background-color:#ff9933"><center>oxidation</center></h2>
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<h2 id="uptake" style="background-color:#ff9933"><center>Uptake</center></h2>
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<h3 style="background-color:#ffdead ">1.Introduction</h3>
<h3 style="background-color:#ffdead ">1.Introduction</h3>
<p>
<p>
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&nbsp;&nbsp;&nbsp;&nbsp;Two proteins called glutathione and thioredoxin which have disulfide bond (-s-s-) in oxidized states exist in E.coli(WT). They play a role of redox control in E.coli.
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<br>&nbsp;&nbsp;&nbsp;&nbsp;Oxidative stress avtivates glutathione reductase (gor) and thioredoxin reductase (trxB). Therefore, E.coli is constantly reductive.<br>
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&nbsp;&nbsp;&nbsp;&nbsp;Fer (Ferric uptake regulator) controls iron metabolism such as taking iron in or storing iron.<br>
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&nbsp;&nbsp;&nbsp;&nbsp;On the other hand, cytosol of yeast is oxidative originally and they can have magnetism, and be attracted by magnets.<br>
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&nbsp;&nbsp;&nbsp;&nbsp;In most cases, it combines with Fur box (which is near the promoter) and regulates transcription.<br>
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&nbsp;&nbsp;&nbsp;&nbsp;So, we thought changing the E.coli cytosol to oxidative state like yeast leads to be attracted by magnets.<br>
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&nbsp;&nbsp;&nbsp;&nbsp;That's why knocking out trxB and gor is neccesary to acchive magnetism.
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&nbsp;&nbsp;&nbsp;&nbsp;When iron is rich, Fur becomes active and when Fur becomes active, it restricts the expression of iron transporter, and that means that the iron uptake would stop. At the same time, Fur restricts the expression of RyhB. RyhB is one of the sRNA that restricts the expression of Ferritin, so making Fur active leads ferritin to express.<br>
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&nbsp;&nbsp;&nbsp;&nbsp;Conversely, when iron is lacking, Fur becomes inactive, and the iron transporter would work normally, so iron would be taken in. RyhB would also work normally, so expression of ferritin is restricted and Ferritin wouldn’t be expressed.<br>
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&nbsp;&nbsp;&nbsp;&nbsp;In short, if we knockdown/knockout Fur, iron transporter would be active so iron would be taken in, but the expression of Ferritin is stopped by RhyB.<br>
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<br>
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&nbsp;&nbsp;&nbsp;&nbsp;Also, there is a Regulator called fief (ferric iron efflux). It makes iron and zinc flow out of cell and controls detoxification of cell. When fieF is knocked down/out, the tolerance of cell to iron would be lowered.<br>
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&nbsp;&nbsp;&nbsp;&nbsp;Fec has a character that can recognize ferric citrate and taking Fe(Ⅲ) in. Fec is controlled by Fur and when the density of Fe(Ⅲ) in the cell is too high, Fur restricts Fec and stops iron uptake.<br>
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&nbsp;&nbsp;&nbsp;&nbsp;In the same way, EfeUOB has a character that can recognize ascorbic acid and taking Fe(Ⅱ) in. EfeUOB is also controlled by Fur and when the density of Fe(Ⅱ) in the cell is too high, Fur restricts EfeUOB and stops iron uptake.<br>
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&nbsp;&nbsp;&nbsp;&nbsp;Using these characters, and by knocking down/out Fur and fieF, Fec/EfeUOB expression wouldn’t be restricted so the amount of iron coming in would increase and the amount of iron going out would decrease. The system would work like when iron is lacking.<br>
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&nbsp;&nbsp;&nbsp;&nbsp;As a result, the amount of  iron inside E.coli would increase.
</p>
</p>
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<h3 style="background-color:#ffdead ">2.Results & Discussion</h3>
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<h3 style="background-color:#ffe4c4 ">2.1.酸化状態を評価</h3>
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<h3 style="background-color:#ffdead ">2.Plasmid</h3>
<p>
<p>
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<a href="#">Assay</a><br>
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プラスミドについて書く<br>
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結果
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<a href="#">Part link</a><br><br>
</p>
</p>
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<h3 style="background-color:#ffe4c4 ">2.2.評価してみて</h3>
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<h3 style="background-color:#ffdead ">3.Results & Discussion</h3>
<p>
<p>
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いろいろわかった
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<a href="#">Assay</a><br>
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実験結果書いていく
</p>
</p>

Revision as of 11:19, 26 September 2013

iGEM-2013 Chiba

iGEM-2013 Chiba

Uptake

1.Introduction

    Fer (Ferric uptake regulator) controls iron metabolism such as taking iron in or storing iron.
    In most cases, it combines with Fur box (which is near the promoter) and regulates transcription.

    When iron is rich, Fur becomes active and when Fur becomes active, it restricts the expression of iron transporter, and that means that the iron uptake would stop. At the same time, Fur restricts the expression of RyhB. RyhB is one of the sRNA that restricts the expression of Ferritin, so making Fur active leads ferritin to express.
    Conversely, when iron is lacking, Fur becomes inactive, and the iron transporter would work normally, so iron would be taken in. RyhB would also work normally, so expression of ferritin is restricted and Ferritin wouldn’t be expressed.

    In short, if we knockdown/knockout Fur, iron transporter would be active so iron would be taken in, but the expression of Ferritin is stopped by RhyB.

    Also, there is a Regulator called fief (ferric iron efflux). It makes iron and zinc flow out of cell and controls detoxification of cell. When fieF is knocked down/out, the tolerance of cell to iron would be lowered.

    Fec has a character that can recognize ferric citrate and taking Fe(Ⅲ) in. Fec is controlled by Fur and when the density of Fe(Ⅲ) in the cell is too high, Fur restricts Fec and stops iron uptake.
    In the same way, EfeUOB has a character that can recognize ascorbic acid and taking Fe(Ⅱ) in. EfeUOB is also controlled by Fur and when the density of Fe(Ⅱ) in the cell is too high, Fur restricts EfeUOB and stops iron uptake.

    Using these characters, and by knocking down/out Fur and fieF, Fec/EfeUOB expression wouldn’t be restricted so the amount of iron coming in would increase and the amount of iron going out would decrease. The system would work like when iron is lacking.

    As a result, the amount of iron inside E.coli would increase.

2.Plasmid

プラスミドについて書く
Part link

3.Results & Discussion

Assay
実験結果書いていく

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