Difference between revisions of "Team:TokyoTech/Experiment/Human Cell Assay"

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<h1 class="w3-xxxlarge w3-text-red" style="padding-bottom: 10px;padding-top: 10px" align="center">Human Cell Assay</h1>
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<h1 class="w3-xxxlarge w3-text-red" style="padding-bottom: 10px;padding-top: 10px" align="center">AHK4 Assay</h1>
 
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     <h1 class="w3-xxxlarge w3-text-red" style="padding-bottom: 10px;padding-top: 10px"><b>Overview</b></h1><!-- 小見出し -->
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     <p style="font-family: Poppins;font-size: 16px">概要をここに
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     <p style="font-family: Poppins;font-size: 16px">In this assay, we checked whether human cells (EAhy926 cell) receive AHL, a signaling molecule derived from <i>E. coli</i> and induce the transcription of <i>atlPT4</i> gene and <i>log1</i> gene which synthesize iP. Also, we checked whether IP (Isopentenyl adenine) molecules are synthesized in EAhy926 cells by TLC.
 
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    <p style="font-family: Poppins;font-size: 10px"><u>Note</u></p>
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    <p style="font-family: Poppins;font-size: 16px">AHLs, which stand for [N-]acyl-homoserine lactones, are small signaling molecules and are employed in the bacterial “Quorum Sensing”. Among several kinds of the signaling molecules, we used 3OC8AHL(C8) in the assay.</p>
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    <p style="font-family: Poppins;font-size: 16px">As shown in Fig. , we introduced two kinds of constructs into EAhy926 cells by electroporation. CAG promoter constantly expresses relA/NLS/traR. After the protein binds with 3OC8AHL(C8), the complex exposes nuclear localization signal and activates CMVmin promoter in CMVmin_atIPT4_IVS_IRES_LOG1_polyA. As a result of pCMVmin's activation, downstream <i>atlPT4</i> gene and <i>log1</i> gene are transcripted. <i>atlPT4</i>  gene and <i>log1</i> gene jointly work as IP synthetase.
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     <h1 class="w3-xxxlarge w3-text-red" style="padding-bottom: 10px;padding-top: 10px"><b>Discussion</b></h1>
 
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     <p style="font-family: Poppins;font-size: 16px">考察
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     <p style="font-family: Poppins;font-size: 16px">We checked the transcription of <i>atlPT4</i> gene and <i>log1</i> gene are induced by C8 addition and the induction depends on C8 concentration.
 
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Revision as of 06:03, 22 October 2017

<!DOCTYPE html> Coli Sapiens

iGEM Tokyo Tech

AHK4 Assay

Introduction

In this assay, we checked whether human cells (EAhy926 cell) receive AHL, a signaling molecule derived from E. coli and induce the transcription of atlPT4 gene and log1 gene which synthesize iP. Also, we checked whether IP (Isopentenyl adenine) molecules are synthesized in EAhy926 cells by TLC.


Note

AHLs, which stand for [N-]acyl-homoserine lactones, are small signaling molecules and are employed in the bacterial “Quorum Sensing”. Among several kinds of the signaling molecules, we used 3OC8AHL(C8) in the assay.

Introduction

As shown in Fig. , we introduced two kinds of constructs into EAhy926 cells by electroporation. CAG promoter constantly expresses relA/NLS/traR. After the protein binds with 3OC8AHL(C8), the complex exposes nuclear localization signal and activates CMVmin promoter in CMVmin_atIPT4_IVS_IRES_LOG1_polyA. As a result of pCMVmin's activation, downstream atlPT4 gene and log1 gene are transcripted. atlPT4 gene and log1 gene jointly work as IP synthetase.

Results

文章

Fig. 画像タイトル
Fig. 画像タイトル

Discussion

We checked the transcription of atlPT4 gene and log1 gene are induced by C8 addition and the induction depends on C8 concentration.

Reference

参考文献

Hajime Fujita: All Rights Reserved