ProjectAims ProjectAims ProjectIdea ROCK SCISSORS PAPER ham - PowerPoint PPT Presentation

Project
Aims


Project
Aims


Project
Idea
 ROCK
 SCISSORS
 PAPER
 ham


‘Rock‐Paper‐Scissors’
Plasmid
Map
 IPTG
 mCherry
 P J23119 P Tac rhIR mCherry lasI P RhI lacI LacI
 Autoinducers
 HSL
 (activates
Plas
in
scissors)
 (from
RhlI
in
paper)
 5


Conway’s
Game
of
Life
 P J23119 P Lux GFP ssRA LuxR LacI ssRA P Tac luxI LacI
 GFP
 AHL
 IPTG
 [Game
of
Life
animation]
http://en.wikipedia.org/wiki/Conway's_Game_of_Life

Modelling


Modelling
  
 Figure
1:
Simulation
results [1]
http://tcl‐nap.sourceforge.net/

Game
of
Life
 Figure
2:
Game
of
Life
results

Rock‐Paper‐Scissors
 Figure
3:
R‐P‐S
beginning
of
simulation

Rock‐Paper‐Scissors
 Figure
4:
R‐P‐S
mid‐simulation

Rock‐Paper‐Scissors
 Figure
5:
R‐P‐S
end
of
simulation

Rock‐Paper‐Scissors
 Figure
6:
Animated
results

Simulator
 [2]
http://2009.igem.org/Team:Southampton/Modeling/Open


Wetlab

 
http://2009.igem.org/Team:Southampton/Project/Safety


Results
‐
Game
of
life
 P J23119 P Lux ssRA ssRA luxR lacI gfp P Tac luxI Original
digestion
 Game of conditions:
2
hours
 Life at
37°C
 Original
ligation
 conditions:
2
hours
 at
RT



Results
‐
Rock,
paper,
scissors
 Original
digestion
 conditions:
2
hours
 at
37°C
 Original
ligation
 conditions:
2
hours
 at
RT

 P J23119 P Tac rhIR mCherry lasI P RhI lacI ROCK

Results
 Original
digestion
 TA
cloning
of
the
 conditions:
2
hours
 insert
followed
by
 at
37°C
 single
digestions

 Original
ligation
 Overnight
gel
 conditions:
2
hours
 puri`ication
 at
RT

 Dephosphorylation
 of
the
vector 


Results
 Original
digestion
 TA
cloning
of
the
 Ligation
 conditions:
2
hours
 insert
followed
by
 conditions:
18
 at
37°C
 single
digestions

 hours
at
16°C
 Original
ligation
 Overnight
gel
 Successful
cloning
 conditions:
2
hours
 puri`ication
 of
P Tac 
 at
RT

 Dephosphorylation
 of
the
vector 


Results
‐
P Tac
 Off No IPTG? Yes LuxI? No Processing No LuxI? Yes On P Tac 
‐
De
Boer
et
al.,
 Proc. Natl. Acad. Sci USA 1983 ,
80,
21‐25


Results
‐
P Tac
 P Tac gfp

Achievements


Christopher Chope MP Alan Whitehead MP

Postgraduate: Stephanie
Chambers
&
Richard
Wilson
 Simula<ons: Paul
Baseo<o
&
Karolina
Kaniewska
 Wet lab: Danielle
Edwards;
Jonathan
Hunter;
 Claire
Sco<;
Jo
Sampson
 &
Tom
Tweed
 Outreach: Charles
Harrison
 Advisors: Prof.
George
A<ard;
Dr
Ali
Tavassoli;
Dr
 Klaus‐Peter
Zauner;
Dr
Marcus
Dymond;
 Dr
Eugen
Stulz
 With thanks also to: Karin
Nordgren ; Dr David Read & Lee Mulholland