Team:UT-Tokyo/Project/Inhibition/System

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このページの概要を、簡単に記述。
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Here we explain the Background & System of Inhibition without Knockout project.
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A repressor normally binds to a promoter and hinders transcription. By introducing extra copies of the repressor binding site in the promoter, the repressor is sequestered by these copies and the probability that the repressor binds to the promoter is decreased. For this reason, the binding of the repressor to the promoter is competitively inhibited by the repressor binding sites.
 
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== 編集の仕方 ==
 
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* ページ右上にあるログインリンクからログインできます。
 
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* ログイン済みの場合は、ページ左上にカーソルを持っていけば、editから内容を編集できます。(日本語メニューの場合は「編集」)
 
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* 新規ページを作るには、アドレスバーに作りたいページのURLを打ち込めばできます。そのページには、このテンプレートページの内容を全てコピーして貼り付け、指定がある部分を編集して自由記述すればOKです。
 
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== wikiの記法 ==
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== Background ==
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In project H<sub>2</sub> ''E.coli'', we tried to improve the formic acid-hydrogen pathway by overexpressing a gene ''fhlA'' which controls a step in this pathway.
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[http://2008.igem.org/Team:Chiba/Internal/foredit 2008年度Team:Chibaのwiki]が参考になります。
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However, ''E.coli'' possess a protein called HycA coded in their genome preventing unrestricted hydrogen synthesis. In order to increase the amount of hydrogen produced, it was thought that there was a need for systems that inhibit the action of negative factors such as HycA.
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そのページでもリンクされていますが、書き方は
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[http://ja.wikipedia.org/wiki/Help:%E3%83%9A%E3%83%BC%E3%82%B8%E3%81%AE%E7%B7%A8%E9%9B%86 Wikipedia-Help:ページの編集]準拠のようです。
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* 表の書き方:[http://ja.wikipedia.org/wiki/Help:%E8%A1%A8%E3%81%AE%E4%BD%9C%E3%82%8A%E6%96%B9 wikipedia Help:表の作り方]
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In general, in order to suppress negative factors such as HycA, a gene knockout method is used. However, because it is easier to introduce a plasmid rather than knocking out a gene, we decided to try to suppress these factors by adding Biobrick Parts. A plasmid containing a large number of repeats of the binding site of the factor is introduced into ''E.coli'' as a decoy to bind the factors. Thus, the factors bind to the decoy Plasmid and we can reduce the binding to its original binding site to inhibit the function. Additionally, by changing the number of decoy binding sites, transcription can be adjusted stepwise.
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== 見出し1 ==
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To this end we used LacI and ArgR whose binding sites are known, and introduced multiple copies of their binding sites into plasmids to examine how the expression levels of genes downstream of these proteins were affected.
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== 見出し2 ==
 
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=== 小見出し1 ===
 
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リンクの例:<br />
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== System ==
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*ページ内リンク
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A repressor normally binds to a promoter and hinders transcription. By introducing extra copies of the repressor binding site in the promoter, the repressor is sequestered by these copies and the probability that the repressor binds to the promoter is decreased. For this reason, the binding of the repressor to the promoter is competitively inhibited by the repressor binding sites.<br/>
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**右のように[[#見出し1]]と書くと見出し1へのリンクが張られます。ページトップ[[#top]]と、各見出しへはこのようにしてリンクが張れます。
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*Wiki内部リンク
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**右のように[[Team:UT-Tokyo/Internal/Sandbox]] と書くとそのまま表示され<br />
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**右のように[[Team:UT-Tokyo/Internal/Sandbox|Sandboxへのリンク]]と書くと「Sandboxへのリンク」という文字列にリンクが張られます。
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*Wiki外部リンク
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**右のようにhttp://2012.igem.org/Team:UT-Tokyoと書くとそのまま表示され、
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**右のように[http://2012.igem.org/Team:UT-Tokyo UT-Tokyoのトップページ]と書くと「UT-Tokyoのトップページ」という文字列にリンクが張られます。
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**右のように[http://2012.igem.org/Team:UT-Tokyo]と書くと、自動で番号のついたリンクが張られます。
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画像にリンクしたい場合:
 
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[[media:example.jpg]]
 
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画像を表示したい場合
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[[File:UT Tokyo Fig comp.inhibit 640 240.png]]
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[[File:ファイル名(拡張子込)]]
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段落内改行は<br />
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;定義リストの定義
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:定義リストの説明
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== 画像類 ==
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[[Team:UT-Tokyo/Internal/Images]]にがぞうのせてる
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Latest revision as of 02:31, 27 September 2012

Inhibition without Knockout:
Background & System

box-background image

Here we explain the Background & System of Inhibition without Knockout project.

Background

In project H2 E.coli, we tried to improve the formic acid-hydrogen pathway by overexpressing a gene fhlA which controls a step in this pathway.

However, E.coli possess a protein called HycA coded in their genome preventing unrestricted hydrogen synthesis. In order to increase the amount of hydrogen produced, it was thought that there was a need for systems that inhibit the action of negative factors such as HycA.

In general, in order to suppress negative factors such as HycA, a gene knockout method is used. However, because it is easier to introduce a plasmid rather than knocking out a gene, we decided to try to suppress these factors by adding Biobrick Parts. A plasmid containing a large number of repeats of the binding site of the factor is introduced into E.coli as a decoy to bind the factors. Thus, the factors bind to the decoy Plasmid and we can reduce the binding to its original binding site to inhibit the function. Additionally, by changing the number of decoy binding sites, transcription can be adjusted stepwise.

To this end we used LacI and ArgR whose binding sites are known, and introduced multiple copies of their binding sites into plasmids to examine how the expression levels of genes downstream of these proteins were affected.


System

A repressor normally binds to a promoter and hinders transcription. By introducing extra copies of the repressor binding site in the promoter, the repressor is sequestered by these copies and the probability that the repressor binds to the promoter is decreased. For this reason, the binding of the repressor to the promoter is competitively inhibited by the repressor binding sites.


UT Tokyo Fig comp.inhibit 640 240.png