Imitations of Gametogenesis & Sexual Reproduction using E. coli

Background

Project description

Project design

Five systems

Results

Modeling

Human practice

Team

Acknowledgements

Background

Project

description

Project design

Five systems

Results

Modeling

Human practice

Team

Ackn.

Imitating developmental process

Trends in Cell Biology, December 2012, Vol. 22, No. 12

Differentiation between Germline and Soma

• The SMB--Synthetic Multicellar Bacterium

Background

Project

description

Project design

Five systems

Results

Modeling

Human

practice

Team

Ackn.

http://2007.igem.org/Paris

Pattern formation Background

Project

description

Project design

Five systems

Results

Modeling

Human

practice

Team

Ackn. Nature 434, 1130-1134 (28 April 2005)

self-organization

http://2008.igem.org/File:USTC_08

_overview.jpg

the combination of the above two

processes

Background

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description

Project design

Five systems

Results

Modeling

Human

practice

Team

Ackn.

Cell migration Background

Project

description

Project design

Five systems

Results

Modeling

Human

practice

Team

Ackn.

http://2011.igem.org/Team:USTC-China

Background

Project

description

Project design

Five systems

Results

Modeling

Human

practice

Team

Ackn.

Background

Project

description

Project design

Five systems

Results

Modeling

Human

practice

Team

Ackn.

Pt7

Ptet lock

activator

Lac+pL

luxR

Lux pR

Lux l

Endogenous promoter

luxpR ftsk Po Flpe-ssrA

Ptet Lux pR

Po T7pol

Fin OP

Cre-ssrA

Pt7

RFP

PtetR Ribokey

Background

Project

description

Project design

Five systems

Results

Modeling

Human

practice

Team

Ackn.

Pt7

GFP-ssrA

Ptet lock

activator

Lac+pL

luxR

Lux+pR

Lux l

Endogenous promoter

luxpR ftsk Flpe-ssrA

Ptet

Lux pR

Po T7pol

Fin OP Cre-ssrA

Pt7

RFP

PtetR Ribokey

Po

Pt7

GFP-ssrA

Ptet lock

activator

Background

Project

description

Project design

Five systems

Results

Modeling

Human

practice

Team

Ackn.

Pt7

GFP-ssrA

Ptet lock

activator Lac+pL

luxR

Lux+pR

Lux l

Endogenous promoter

luxpR ftsk Flpe-ssrA

Ptet

Lux pR

Po T7pol

Fin OP Cre-ssrA

Pt7

RFP

PtetR Ribokey

Po

Background

Project

description

Project design

Five systems

Results

Modeling

Human

practice

Team

Ackn.

Pt7

GFP-ssrA

Ptet lock

activator

Lac+pL

luxR

Lux+pR

Lux l

Endogenous promoter

luxpR ftsk Flpe-ssrA

Ptet

Lux pR

Po T7pol

Fin OP Cre-ssrA

Pt7

RFP

PtetR Ribokey

Po

a) Quorum Sensing system in high copy plasmid;

b) Controlled DNA invertase device insertted in genome;

c) Real-time fluorescence reporters;

d) Three regulators and its responding promoters;

e) F factor and its incompatible plasmid

Background

Project

description

Project design

Five systems

Results

Modeling

Human

practice

Team

Ackn.

lacl+pl

luxr

Lux pR

Five systems a) Quorum Sensing

system in high copy plasmid;

b) Controlled DNA invertase device insertted in genome;

c) Real-time fluorescence reporters;

d) Three regulators and its responding promoters;

e) F factor and its incompatible plasmid

Five systems a) Quorum Sensing

system in high copy plasmid;

b) Controlled DNA invertase device insertted in genome;

c) Real-time fluorescence reporters;

d) Three regulators and its responding promoters;

e) F factor and its incompatible plasmid

PO

Endogenous promoter ftsk Lux-pR Flpe-ssrA

Lox

Lox

T7 pol

P(tet)

PO

Lux-pR

creCre-ssrA PO Lux-pR

P(tet)

PO

RFP P(lac) Cre-ssrA

GFP-ssrA

Five systems a) Quorum Sensing

system in high copy plasmid;

b) Controlled DNA invertase device insertted in genome;

c) Real-time fluorescence reporters;

d) Three regulators and its responding promoters;

e) F factor and its incompatible plasmid

Five systems a) Quorum Sensing

system in high copy plasmid;

b) Controlled DNA invertase device insertted in genome;

c) Real-time fluorescence reporters;

d) Three regulators and its responding promoters;

e) F factor and its incompatible plasmid

GFP-ssrA Pt7

Three regulators and its responding promoters

.

a) T7 polymerase and T7 promoter

b) Riboregulator

c) Delta activator from phiR73 phage

Five systems a) Quorum Sensing

system in high copy plasmid;

b) Controlled DNA invertase device insertted in genome;

c) Real-time fluorescence reporters;

d) Three regulators and its responding promoters;

e) F factor and its incompatible plasmid

b) Riboregulator Five systems a) Quorum Sensing

system in high copy plasmid;

b) Controlled DNA invertase device insertted in genome;

c) Real-time fluorescence reporters;

d) Three regulators and its responding promoters;

e) F factor and its incompatible plasmid

a) T7 polymerase and T7 promoter

P(tet) R0040 J23008 J23009 B0010 B0012

P(tet) Lock P(tet) J23032

Negative control: BBa_J61046

16 16.5 17 17.5 18 18.5 190

5

10

15

20

25

hours

Flu

ore

scence q

uantification/O

D v

alu

e

6I+3N+20L

6I+3N+20L+13J

5H

Figure 1. fluorescence observation using fluorescence microscope

Figure2. quantitatively measurement of the fluorescence intensity.

Positive control

R0040 J23032

BBa_K1087015

mRFP1

BBa_K1087021 R0040 mRFP1E

1010 B0034

R0040 J23008 J23009 J23032 R0040 mRFP1

Ribolock

Positive

Ribokey+Ribolock

Delta activator from phiR73 phage

BBa_K1087018

BBa_K1087022

BBa_K1087023

A B

C D

A B

C D

Figure 1. fluorescence observation using fluorescence microscope

Figure2. quantitatively measurement of the fluorescence intensity.

16 16.5 17 17.5 18 18.5 190

10

20

30

40

50

60

70

hours

Flu

ore

scence q

uantification/O

D v

alu

e

7H+23L+13J

7H+22O+6I~23L

7H+22O

Ribokey+activator

Basal level

Ribokey+activator

Five systems a) Quorum Sensing

system in high copy plasmid;

b) Controlled DNA invertase device insertted in genome;

c) Real-time fluorescence reporters;

d) Three regulators and its responding promoters;

e) F factor and its incompatible plasmid

logistic growth model

cre concentration

Background

Project

description

Project design

Five systems

Results

Modeling

Human

practice

Team

Acknowledgem

ents

Model 1 Transforming into gametes

)1(max

1N

NNk

dt

dN

ttT

Cdt

cd

tttT

CNkdt

cd

crecre

cresignccre

2

21

5.0ln

)(

5.0ln

)(

Model 1 Figure

Background

Project

description

Project design

Five systems

Results

Modeling

Human

practice

Team

Ackn.

Model 1 Figure Background

Project

description

Project design

Five systems

Results

Modeling

Human

practice

Team

Acknowledgem

ents

Lindbergh - Levi central limit theorem

Step 1.Calculating the sample's

expectations E(ξ) =μ, variance D (ξ) =σ2

and standardization

Step 2 By the Lindbergh - Levi central limit

theorem

we get function of concentration:

Background

Project

description

Project design

Five systems

Results

Modeling

Human

practice

Team

Acknowledgem

ents

)(1

1

k

N

kN

N

dte2

1)(lim

x

-

2

t2

πxP N

N

}

)(

{lim)( 1 xN

Px

N

k

k

N

Model 2:Gametes are combined

• Markov random modeling

Background

Project

description

Project design

Five systems

Results

Modeling

Human

practice

Team

Acknowledgem

ents

])1(x)(x[

])1-n(x,...,)1(x,)0(x)([

)1(

)1(10

nn

nn

ininP

iiiinxP

丨

丨

1. SCU Synthetic Biology Association

2. Synthetic Biology Home

Background

Project

description

Project design

Five systems

Results

Modeling

Human

practice

Team

Ackn.

http://www.isynbio.com/forum.php

Advisor and team members Background

Project

description

Project design

Five systems

Results

Modeling

Human

practice

Team

Ackn.

• Sichuan University, SCU

• Academic Affairs Office, SCU

• School of Life Sciences, SCU

Background

Project

description

Project design

Five systems

Results

Modeling

Human

practice

Team

Ackn.