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- 1. Why “E.coli Pen ”
- 2. Experiments
- 3. How to use
2. Experiments 3. How to use Q. What do you think of the artificial - - PowerPoint PPT Presentation
2. Experiments 3. How to use Q. What do you think of the artificial - - PowerPoint PPT Presentation
E.Coli pen Draw with your own color. Todays 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
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- Q. What do you think of the
artificial genetic mutation?
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66%
32% 2%
Bad Good No respons
n=1284
Negative feeling for Biotechnology and Genetic Engineering
Results of iGEM JAPAN Survey
No response
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A r t Science
Enjoy and understand the biotechnology
Bio Art
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A r t Science
Science and Art meet in Kyoto Art Science
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Previous Bio Art
Only in laboratories Need equipments
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New Bio Art Tool E.Coli Pen
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New Bio Art Tool E.Coli Pen
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Theory of three primary colors
Promoter 1 RFP Promoter 2 GFP Promoter 3 CFP CFP GFP RFP
To make various inks
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- 1. Why “E.coli Pen ”
- 2. Experiments
- 3. How to use
Today’s topics
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500 1000 1500 2000 2500 3000
pSB6A1 (Low Copy Vector) pSB1A2 (High Copy Vector) Fluorescent density
[%]
select
The best vector for GFP expression
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The best promoters for E.coli Pen
Controlled by easily available substances
Different sensitivity to substances
Not used in previous iGEM
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Oxidative stress
Cell death
Cancer
Promoters responding to H2O2
Diabetes Hypertension Arteriosclerosis Wrinkle Aging etc...
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Survival ratio of E.coli under H202 stress
We used concentration of H202 , from 1 nM to 1 mM
50 100 150 200 250 0.000001 0.0001 0.01 1 100
H202 concentration Number of colonies/plate
[mM]
1 nM 1 mM
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Seven promoters responsive to H2O2
Inactive form Active form H2O2
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sodA ahpC sufA acrAB
- xyR
dps yaiA Promoters Binding
Seven promoters responsive to H2O2
Transcription… Active form
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ahpC sufA We have selected ahpC and sufA promoters.
Seven promoters responsive to H2O2
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OxyR activates the transcription of ahp operon
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5 10 15 20 25 30 10 20 30 40 50 60 70 80 less 1nM 10nM 100nM 1µM 10µM 100µM 1mM
(min.)
H2O2
ahpC promoter activity
Green Fluorescence/O.D.(488 nm)
1 mM 10 µM No treatment
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- xyR
H2O2
suf A suf B suf C suf S suf E
Reduced form Oxidized form Dimer OxyR binds to the OBR activates the transcription of the suf operon
OxyR activates the transcription of suf operon
OBR OBR
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0.5 1 1.5 2 2.5 3 3.5 4 4.5 5 10 20 30 40 50 60 70 80 90 less 1nM 10nM 100nM 1µM 10µM 100µM 1mM
H2O2
Depends on Concentration
1 mM 100 µM 10 µM
No treatment
sufA promoter activity
Green Fluorescence/O.D.(488 nm)
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Making inks
RFP : GFP = 2 : 1 RFP : GFP = 1 : 2 GFP Control (PBS) RFP
Promoter Promoter
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GFP
sufA
GFP
ahpC
GFP
acrAB
GFP
dps
GFP
- xyR
GFP
sodA
GFP
yaiA
CFP CFP CFP CFP
We designed many parts
YFP RFP CFP YFP RFP YFP RFP YFP RFP CFP CFP YFP RFP YFP RFP YFP RFP
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- 1. Why “E.coli Pen ”
- 2. Experiments
- 3. How to use
Today’s topics
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How to use
the ‘‘E.coli pen’’
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1
Make your own color.
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Select the tubes ‘‘ E.coli ’’ or ‘‘ H2O2 ’’.
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Knock the pen and make color.
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Under the black light
Make color as you like.
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Anti-contamination system
to keep the color
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2
Draw with your own color!
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Finished! Un
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3
Irradiated with the black light.
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Irradiate your works.
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fin.
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Very important item to enjoy Bio Art
?
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Pinch?
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How to use
This
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- hint. E.coli smells not good...
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Like this!
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Thank you for your attention !
Our sponsors
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