RegionalTeamMeetingsand Bootcamp Fiveschoolsatthreecampuses - PowerPoint PPT Presentation

Regional
Team
Meetings
and
 Bootcamp 
 • Five
schools
at
three
campuses
 • Skype,
email,
Google
wave,
and
regional
meetings
used
to
 communicate
between
campuses.
 • Boot
camp
held
at
beginning
of
summer
to
bring
each
team
 together.


Quorum
Sensing
Amplifiers
 and
a
Co‐design
Approach
 for
Information
Processing 
 “Ditoxotector:
 Dual
Toxin
Detection”


Motivation 
 Applications
 Effect
 Fish
‐
Most
heavy
metal
 Environment
 combinations
 Human
liver
‐
Carbon
 Health
 Tetrachloride
and
Ethanol


Toxins
alone… 
 As Hg

…but
with
two… 
 As Hg

Goals 
 • Bacterial
biosensor
detects
simultaneous
 presence
of
two
toxins
 • Modular
design
allows
for
extension/
 improvement


Project
Overview 
 Input Amplification Logic/ Output Mercury Arsenic Copper

Toxic
Metal
Detection 


Arsenic
Switch
 ArsR Promoter Operator

Arsenic
Switch
 As As ArsR ArsR ArsR GFP GFP Promoter Operator Operator

Project
Overview 
 Input Amplification Logic/ Output Mercury Arsenic Copper

Quorum
Sensing
Design 


Amplifier
Model 
 Without the amplifier output is driven directly by the toxin switch. With the amplifier output is driven by the dimer. Dimer production is driven by the toxin switch as well as by positive feedback

Amplifier
Model 


Project
Overview 
 Input Amplification Logic/ Output Mercury Arsenic Copper

Logic
Structure 
 Input
1
 Input

2
 Input
3
 Output
 Output
 Output
 (Hg)
 (Cu)
 (Ar)
 Signal
A
 Signal
B
 Signal
C
 Detected
 ‐
 ‐
 ‐
 ‐
 ‐
 ‐
 Detected
 ‐
 ‐
 ‐
 ‐
 ‐
 ‐
 Detected
 ‐
 ‐
 ‐
 Detected
 Detected
 ‐
 Emitted
 ‐
 ‐
 Detected
 ‐
 Detected
 ‐
 Emitted
 ‐
 ‐
 Detected
 Detected
 ‐
 ‐
 Emitted
 Detected
 Detected
 Detected
 Emitted
 Emitted
 Emitted


Methods
for
AND
Logic
 A
 B
 Hybrid
Promoters
 Fluorescent
Complementation


Hybrid
Promoters
 • Constitutive
Promoters
J23110
&
J23106
 • Combination
of
2
operator
sites

 Tet Operator Lac Operator -35 -10 Ara Operator Tet Operator -35 -10 Ara Operator Lac Operator -35 -10

Our
System 


Complementation
 Magliery et. al 2005

Putting
it
all
together 
 • Co‐Transformation
 • Three
Antibiotic
 Resistances
per
 Plasmid
 Top10 E. coli cotransformed with pUC19 and pSB2K3 plasmids.

Optimization
 Used
for
creation
of
 customized
RBS
 sequences
for
the
PCR
 cassettes

 Used
to
evaluate
the
 effectiveness
of
the
current
 BioBrick
fluorescent
protein
 cassettes


RBS (only the Shine-Dalgarno) AAAGAGGAGAAA 11564.6 378.2

Possible
Applications 
 • Environmental
 ‐
Replace
fluorescence
with
remediation
 • National
Security
 ‐
Bomb
–
Thiodigylcol
and
Phosphorous
 Trichloride
(Mustard
Gas)


Conclusions 
 • Design
of
Quorum
Sensing
Amplifier
 • Construction
of
Hybrid
Promoters
 • Design
of
Complementation
System



iGEM
Experience 
 • Regional
team
experiment
itself
 • Influence
on
project
 • Learning
to
communicate
and
work
together
 • Vital
experiences
to
the
future
of
synthetic
 biology


Acknowledgements
 
 Our
Advisors 
 National
Science
Foundation 
 Mr.
Gene 
 Bluefield
State
College