E.Coli pen Draw with your own color.
Today’s topics 1. Why “ E.coli Pen ” 2. Experiments 3. How to use
Q. What do you think of the artificial genetic mutation?
Results of iGEM JAPAN Survey 2% 32% Bad 66% Good No respons No response n=1284 Negative feeling for Biotechnology and Genetic Engineering
A r t Science Bio Art Enjoy and understand the biotechnology
Art Science A r t Science Science and Art meet in Kyoto
Previous Bio Art Only in laboratories Need equipments
New Bio Art Tool E.Coli Pen
New Bio Art Tool E.Coli Pen
To make various inks Theory of three primary colors Promoter 1 RFP Promoter 2 GFP Promoter 3 CFP RFP GFP CFP
Today’s topics 1. Why “ E.coli Pen ” 2. Experiments 3. How to use
The best vector for GFP expression [%] 3000 Fluorescent density 2500 2000 1500 select 1000 500 0 pSB1A2 pSB6A1 (High Copy Vector) (Low Copy Vector)
The best promoters for E.coli Pen Controlled by easily available substances Different sensitivity to substances Not used in previous iGEM
Promoters responding to H 2 O 2 Cancer Oxidative stress Diabetes Hypertension Arteriosclerosis Wrinkle Aging etc... Cell death
Survival ratio of E.coli under H 2 0 2 stress 250 Number of colonies/plate 200 150 100 50 1 mM 1 nM 0 [mM] 0.000001 0.0001 0.01 1 100 H 2 0 2 concentration We used concentration of H 2 0 2 , from 1 nM to 1 mM
Seven promoters responsive to H 2 O 2 H 2 O 2 Inactive form Active form
Seven promoters responsive to H 2 O 2 acrAB sodA ahpC sufA Active form Transcription… yaiA Binding oxyR dps Promoters
Seven promoters responsive to H 2 O 2 ahpC sufA We have selected ahpC and sufA promoters.
OxyR activates the transcription of ahp operon
ahpC promoter activity 30 Green Fluorescence/O.D.(488 nm) 1 mM H 2 O 2 25 No treatment less 20 1nM 10 µM 10nM 15 100nM 1µM 10 10µM 100µM 5 1mM 0 (min.) 10 20 30 40 50 60 70 80
OxyR activates the transcription of suf operon oxyR H 2 O 2 Reduced form Oxidized form Dimer activates the transcription of the suf operon OBR suf A suf B suf C suf S suf E OBR OxyR binds to the OBR
sufA promoter activity Green Fluorescence/O.D.(488 nm) 5 H 2 O 2 1 mM 4.5 Depends on Concentration No treatment 4 less 3.5 1nM 100 µM 3 10nM 2.5 100nM 10 µM 2 1µM 1.5 10µM 1 100µM 0.5 1mM 0 10 20 30 40 50 60 70 80 90
Making inks Promoter Promoter RFP GFP RFP : GFP = 2 : 1 Control (PBS) RFP : GFP = 1 : 2
We designed many parts YFP RFP CFP GFP sufA RFP CFP YFP GFP ahpC YFP RFP GFP CFP acrAB CFP YFP RFP GFP dps YFP RFP GFP CFP oxyR YFP RFP GFP CFP sodA GFP YFP RFP CFP yaiA
Today’s topics 1. Why “ E.coli Pen ” 2. Experiments 3. How to use
How to use the ‘‘ E.coli pen’’
1 Make your own color.
Select the tubes ‘‘ E.coli ’’ or ‘‘ H 2 O 2 ’’.
Knock the pen and make color.
Make color as you like. Under the black light
Anti-contamination system to keep the color
2 Draw with your own color!
Un Finished!
3 Irradiated with the black light.
Irradiate your works.
fin.
? Very important item to enjoy Bio Art
Pinch?
How to use This
hint. E.coli smells not good...
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