Transcript System Design

MIT’s Eau d’e coli iGEM 2006
By: Robert Ovadia
1
Background
System Design
Results
What I Liked and Disliked
Background
What makes a scent?
An enzyme that converts something
that DOESN’T smell, into something
that DOES smell.
2
JMT
CCMT
BAMT
BSMT
ATF1
jasmine
cinnamon
fresh / floral
wintergreen
banana
Background
System Design
Results
What I Liked and Disliked
3
Background
BSMT
ATF1
wintergreen
banana
Background
System Design
• WGD
• BGD
• SAGD
• IAGD
• pOsmY + Inverter
Results
What I Liked and Disliked
System Design
QuickTime™ and a
TIFF (LZW) decompressor
are needed to see this picture.
4
Background
System Design
• WGD
• BGD
• SAGD
• IAGD
• pOsmY + Inverter
Results
What I Liked and Disliked
System Design
Simple System
QuickTime™ and a
TIFF (LZW) decompressor
are needed to see this picture.
5
Background
System Design
• WGD
• BGD
• SAGD
• IAGD
• pOsmY + Inverter
Results
What I Liked and Disliked
System Design
Complex System
QuickTime™ and a
TIFF (LZW) decompressor
are needed to see this picture.
6
Background
System Design
• WGD
• BGD
• SAGD
• IAGD
• pOsmY + Inverter
Results
What I Liked and Disliked
System Design
More Complex System
QuickTime™ and a
TIFF (LZW) decompressor
are needed to see this picture.
7
Background
System Design
• WGD
• BGD
• SAGD
• IAGD
• pOsmY + Inverter
Results
What I Liked and Disliked
System Design
Complete System
QuickTime™ and a
TIFF (LZW) decompressor
are needed to see this picture.
8
Background
System Design
• WGD
• BGD
• SAGD
• IAGD
• pOsmY + Inverter
Results
What I Liked and Disliked
System Design
Wintergreen Generating Device (WGD)
QuickTime™ and a
TIFF (LZW) decompressor
are needed to see this picture.
9
Background
System Design
• WGD
• BGD
• SAGD
• IAGD
• pOsmY + Inverter
Results
What I Liked and Disliked
System Design
Banana Generating Device (BGD)
QuickTime™ and a
TIFF (LZW) decompressor
are needed to see this picture.
10
Background
System Design
• WGD
• BGD
• SAGD
• IAGD
• pOsmY + Inverter
Results
What I Liked and Disliked
System Design
Salicylic Acid Generating Device (SAGD)
QuickTime™ and a
TIFF (LZW) decompressor
are needed to see this picture.
11
Background
System Design
• WGD
• BGD
• SAGD
• IAGD
• pOsmY + Inverter
Results
What I Liked and Disliked
System Design
Isoamyl Alcohol Generating Device (IAGD)
QuickTime™ and a
TIFF (LZW) decompressor
are needed to see this picture.
12
Background
System Design
• WGD
• BGD
• SAGD
• IAGD
• pOsmY + Inverter
Results
What I Liked and Disliked
System Design
OsmY + Inverter
osmY: active in stationary phase & under high osmotic pressure conditions
13
osm
Y
osmY + inverter
Background
System Design
Results
• WGD
• Growth Dependent
Regulation
• Growth Dependent
Regulation + Inverter
• Indole Free Bacteria
• Smell Test
What I Liked and Disliked
Results
Wintergreen Generating Device
E. coli
no methyl salicylate
[no peak at 9.1
minutes]
E. coli + precursor
(SA) + WGD
methyl salicylate
MW = 152
O
O
OH
14
Background
System Design
Results
• WGD
• Growth Dependent
Regulation
• Growth Dependent
Regulation + Inverter
• Indole Free Bacteria
• Smell Test
What I Liked and Disliked
Results
Growth Dependent Regulation
GFP Signal (relative units)
2E+5
BBa_J45995 (pOsmY-GFP)
2E+5
BBa_J45017 (Negative control)
2E+5
BBa_I7100 (Constitutive promoter-GFP)
1E+5
1E+5
1E+5
8E+4
6E+4
4E+4
2E+4
0E+0
0.0
0.1
0.2
0.3
0.4
Absorbance 600nm (relative units)
15
0.5
Background
System Design
Results
• WGD
• Growth Dependent
Regulation
• Growth Dependent
Regulation + Inverter
• Indole Free Bacteria
• Smell Test
What I Liked and Disliked
Results
Growth Dependent Regulation + Inverter
16
Background
System Design
Results
• WGD
• Growth Dependent
Regulation
• Growth Dependent
Regulation + Inverter
• Indole-Free Bacteria
• Smell Test
What I Liked and Disliked
wild type E. coli
Results
Indole-Free Bacteria
chassis: tnaA5- E. coli
Indole
Peak at 10 minutes
MW = 117
No indole
[no peak at 10
minutes]
17
Background
System Design
Results
• WGD
• Growth Dependent
Regulation
• Growth Dependent
Regulation + Inverter
• Indole-Free Bacteria
• Smell Test
What I Liked and Disliked
Results
Smell Test
100
90
80
Percentage
70
60
Banana
Mint
Stinky
50
40
30
20
10
0
A (Mint)
18
B (Stinky)
C (Banana)
Background
System Design
Results
What I Liked and Disliked
• Likes
What I Liked
Likes
•Able to change a simple experiment into a more
complicated experiment
•They looked at what MORE there was to do; took their
experiment to the ‘next level’ EX: Simple System to
Complete System
•Got the audience involved, EX: Smell test
•Easy to comprehend
•Perfect example of a successful iGEM project
19
Sources
MIT’s iGEM PPT Presentation 2006.ppt (PowerPoint)
MIT’s iGEM PPT Presentation 2006.mov (Movie)
http://openwetware.org/wiki/IGEM:MIT/2006
20