colorful e.coli weave time and space2011.igem.org/files/presentation/whu-china_championship.pdf ·...

Colorful E.coli Weave Time and Space

iGEM_Team_11 WHU_China

Wuhan_University

Our University and Team

Magic: Sichuan Opera

Sichuan Opera is a “ national treasure” in China.

Oscillator（time）

Time

Y

X

E.coli Color Film (space)

Colorful E.coli Weave Time and Space

New Standards

Oscillator

How do E.coli Weave Time？

Reporter

GFP LsrR LsrK cI pLsrA/cI pLsrA

B2 B1

Oscillator: Gene Circuit Design

Time

Signal

LsrR LsrK

Design

Model

• The ODE(Ordinary Differential Equation):

Oscillator: Equation

1 2

1 1 2 2

3

3 3

1 2 11 1 1 2 1

1 2 2 1

2 13 2 2 2

2 1

(1 )

(1 )

n n

n n n n

n

n n

dx x xa b x a x

dt k x k x

dx xa x b x

dt k x

LsrR LsrK

0 5 10 15 20 25 30 35 40 45 500

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

Time

Concentr

ation

x1

x2

Deterministic Oscillations

0 5 10 15 20 25 30 35 40 45 500

100

200

300

400

500

600

700

800

TimeN

um

ber

of

mole

cule

s

x1

x2

Stochastic Oscillations

Oscillator: Simulation Diagram

LsrR LsrK

Oscillator: Bifurcation Diagram

a1 b2

X2 X2

0.8

0.6

0.4

0.2

0 0

0.05 0.1 0.15 0.2 0.25 0.35 0.3 0.4 0.5 0.45

0.8

0.6

0.4

0.2

0 0

0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 1.8 2.0

X2 X2

0.8

0.6

0.4

0.2

0

0

1 2 3 4 5 6 a2

0.8

0.6

0.4

0.2

0 0

2 10 4 12 6 14 8 a3

Oscillator: Period Variation with Parameters

8

7

6

5

4

3

2

1

0 0.05 0.1 0.15 0.25 0.2 0.3 0.4 0.35 0.45

30

25

20

15

10

5

0 0.4 0.2 0.8 0.6 1 1.4 1.2 1.8 1.6 2

Period Period

0 0 0.5

10

8

6

4

2

0

0 1 2 4 5 6 a2

3

6

5

4

3

2

1

0 0

2 4 6 10 12 14 a3 8

X2 X2 a1 b2

Conclusion: Colorful E.coli Weave Time and Space

Time

Y

E.coli Color Film (space)

X

Oscillator（time）

Constructed mathematical models to analyze our oscillator.

0 5 10 15 20 25 30 35 40 45 500

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

Time

Concentr

ation

x1

x2

New Standards

E.coli Color Film

How do E.coli Weave Space？

Color Film: Advantages of E.coli Color Film

Traditional film E.coli color film

Complex Convenient

Expensive Cost less

Harmful Harmless

Pollution Environment friendly

Low resolution High resolution

28 600W 4000W

• How to construct?

• Light tricolor theory: red, green and blue can compose any light with different proportion.

• Also the complementary colors which are cyan, yellow and magenta can also compose any color with different proprtion.

Color Film: Mechanism of E.coli Color Film

Blue strain

• Blue light sensor

• Yellow pigment reporting system

Green strain

• Green light sensor

• Magenta pigment reporting system

Red strain

• Red light sensor

• Cyan pigment reporting system

Color Film: Design of E.coli Color Film

Color Film: Design of E.coli Color Film

Orange Luciferase Lov/TAP

Max 450nm

pTrp

Blue strain

• Blue light sensor

• Yellow pigment reporting system

Color Film: Design of Blue Strain

Blue Light Dark

Color Film: Results of Blue Light Sensor

Color Film: Results of Blue Light Sensor

Light sensor

CI

Pigment LuxR

B0034 C0051 B0010 B0012

K145150 B0034 C0062 B0034 Pigment coding gene

K091143 B0034 C0079 B0030 C0053 B0010 B0012

Color Film: Future Work—Amplify System

AHL

Light sensor

CI

LasR CII

Pigment LuxR

AHL

B0034 C0051 B0010 B0012

K145150 B0034 C0062 B0034 Pigment coding gene

K091143 B0034 C0079 B0030 C0053 B0010 B0012

Color Film: Future Work—Amplify System

Conclusion: Colorful E.coli Weave Time and Space

Time

Y

E.coli Color Film (space)

X

Oscillator（time）

Constructed one strain E.coli of sensing blue light. New Standards

Application

E.coli Color Film

E.coli Weave Space.

Oscillator

E.coli Weave Time.

Colorful logo Auto-changing mask

Future Work: Application

Future Work: Application

Smart Lamp Post Tree

Lizard+ Colorful E.coli =Chameleon!

People+ Colorful E.coli= Cloak of Invisibility

Future Work: Application

Conclusion: Colorful E.coli Weave Time and Space

Time

Y

E.coli Color Film (space)

X

Oscillator（time）

Constructed one strain E.coli of sensing blue light.

Constructed mathematical models to analyze our oscillator.

0 5 10 15 20 25 30 35 40 45 500

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

Time

Concentr

ation

x1

x2

New Standards

Biobricks Assembly

New Standards

Reverse Assembly

Golden Gate Method

New Standards

CHL

Tet

Tet

Tet

Tet

Amp

Amp

Amp

Amp

ATGC GTAC

GGTC ATGC

CCGC GCTT

GTAC CTAT Part1

Part2

Part3

Part 10

Golden Gate Method

Golden Gate Method

Traditional Assembly Golden Gate Cloning

Part 2

Amp

E X HindIII

S P HindIII

Part 1

Amp

E X HindIII

S P HindIII

Part 1

Amp

E X HindIII

S P HindIII

Part 2

HindIII

Part 1

HindIII HindIII

Part 1

Amp

E X HindIII

S P HindIII

part2 part1

part2 part1

part2 part1

Reverse Assembly

Summary: Colorful E.coli Weave Time and Space

Time

Y

E.coli Color Film (space)

X

Oscillator（time）

Constructed one strain E.coli of sensing blue light.

X Golden Gate Method; Reverse Assembly

Constructed mathematical models to analyze our oscillator.

0 5 10 15 20 25 30 35 40 45 500

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

Time

Concentr

ation

x1

x2

New Standards

Instructors: Prof Jiangbo Wang Prof Zhixiong Xie

Acknowledgement

Advisors: Prof Lin Guo Prof Xiaodong Zhang Yan Long