Difference between revisions of "Team:TokyoTech/Experiment/Human Cell Assay"
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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. | <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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Revision as of 06:05, 22 October 2017
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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.
Summary
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
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
