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Revision as of 09:14, 31 October 2017

<!DOCTYPE html> Coli Sapiens

iGEM Tokyo Tech

InterLab

Introduction

In iGEM 2017, the InterLab Study is one of the bronze medal criteria. We measured the GFP expression level of the six designated devices, positive control and negative control using the plate reader. It was the second time for our team to join this InterLab Study (The first time in iGEM 2015).


Summary

Our purpose was to investigate the RBS intensity and promoter intensity effects on gene expression by obtaining the fluorescence data of samples and comparing them. We prepared Device1〜Device6, Positive Control and Negative Control as shown below.


Fig. 1 Designated devices

Results

1. We obtained LUDOX correction values.


Fig. 2 LUDOX correct value

2. We obtained the two Fluorescein Standard Curve (normal and log scale).


Fig. 3 Fluorescein Standard Curve
Fig. 4 Fluorescein Standard Curve (log scale)

3. We obtained the graph bellow. The RFU of GFP / Turbidity was in the following order, Device1>Device4>Device2>Positive Control>Device5>Device3>Device6>Negative Control.


Fig. 5 Fluorescein (using AVERGE)
Fig. 6 Fluorescein (using GEOMEAN)

Discussion

Judging from the results, the gene expression is controlled by the intensity of promoter and the intensity of RBS. The promoter intensity of is supposed to be in the following order, J23101>J23106>J23117. The RBS intensity of I13504 is supposed to be higher than that of BCD2.E0040. Moreover, it is considered that the GFP expression level is dependent on individual to some extent.


Materials and Methods

Hajime Fujita: All Rights Reserved