ProjectAims ProjectAims ProjectIdea ROCK SCISSORS PAPER ham - - PowerPoint PPT Presentation

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ProjectAims ProjectAims ProjectIdea ROCK SCISSORS PAPER ham - - PowerPoint PPT Presentation

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ProjectAims ProjectAims ProjectIdea ROCK SCISSORS PAPER ham RockPaperScissorsPlasmidMap IPTG mCherry P J23119 P Tac rhIR mCherry lasI P RhI lacI LacI Autoinducers HSL

slide-1
SLIDE 1
slide-2
SLIDE 2

Project
Aims


slide-3
SLIDE 3

Project
Aims


slide-4
SLIDE 4

ham


ROCK
 PAPER
 SCISSORS


Project
Idea


slide-5
SLIDE 5

5


PJ23119 PTac mCherry lasI rhIR PRhI lacI

IPTG
 HSL
 (from
RhlI
in
paper)
 LacI
 mCherry
 Autoinducers
 (activates
Plas
in
scissors)


‘Rock‐Paper‐Scissors’
Plasmid
Map


slide-6
SLIDE 6

Conway’s
Game
of
Life


PJ23119 LacI LuxR PTac luxI ssRA ssRA PLux GFP

IPTG
 LacI
 GFP
 AHL


[Game
of
Life
animation]
http://en.wikipedia.org/wiki/Conway's_Game_of_Life

slide-7
SLIDE 7

Modelling


slide-8
SLIDE 8

Modelling


[1]
http://tcl‐nap.sourceforge.net/

Figure
1:
Simulation
results

slide-9
SLIDE 9

Game
of
Life


Figure
2:
Game
of
Life
results

slide-10
SLIDE 10

Rock‐Paper‐Scissors


Figure
3:
R‐P‐S
beginning
of
simulation

slide-11
SLIDE 11

Rock‐Paper‐Scissors


Figure
4:
R‐P‐S
mid‐simulation

slide-12
SLIDE 12

Rock‐Paper‐Scissors


Figure
5:
R‐P‐S
end
of
simulation

slide-13
SLIDE 13

Rock‐Paper‐Scissors


Figure
6:
Animated
results

slide-14
SLIDE 14

Simulator


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


slide-15
SLIDE 15

Wetlab




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


slide-16
SLIDE 16

Results
‐
Game
of
life


Original
digestion
 conditions:
2
hours
 at
37°C
 Original
ligation
 conditions:
2
hours
 at
RT



lacI PLux luxI PTac Game of Life luxR PJ23119 gfp ssRA ssRA

slide-17
SLIDE 17

Results
‐
Rock,
paper,
scissors


Original
digestion
 conditions:
2
hours
 at
37°C
 Original
ligation
 conditions:
2
hours
 at
RT

 rhIR

PJ23119

mCherry

PTac

lasI lacI

PRhI

ROCK

slide-18
SLIDE 18

Original
digestion
 conditions:
2
hours
 at
37°C
 Original
ligation
 conditions:
2
hours
 at
RT

 TA
cloning
of
the
 insert
followed
by
 single
digestions

 Overnight
gel
 puri`ication
 Dephosphorylation


  • f
the
vector


Results


slide-19
SLIDE 19

Results


Original
digestion
 conditions:
2
hours
 at
37°C
 Original
ligation
 conditions:
2
hours
 at
RT

 TA
cloning
of
the
 insert
followed
by
 single
digestions

 Overnight
gel
 puri`ication
 Dephosphorylation


  • f
the
vector


Ligation
 conditions:
18
 hours
at
16°C
 Successful
cloning


  • f
PTac

slide-20
SLIDE 20

Results
‐
PTac


Off

IPTG?

Processing

LuxI?

On

LuxI? No

No No Yes Yes

PTac
‐
De
Boer
et
al.,
Proc. Natl. Acad. Sci USA 1983,
80,
21‐25


slide-21
SLIDE 21

Results
‐
PTac


gfp

PTac

slide-22
SLIDE 22

Achievements


slide-23
SLIDE 23
slide-24
SLIDE 24

Alan Whitehead MP Christopher Chope MP

slide-25
SLIDE 25
slide-26
SLIDE 26

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