20 12-iGEM

2012-iGEMBreak Symmetry

Southeast university

凌昕彤

周醒驭

陈牧原

高 珅

Team members

周 乐 程 鹏

桂 凯

张 俊 樊雪龙

徐小丹

贾正阳

Technical Background

Design Scheme

Results && Biobricks &&Potential Work

ContentsContents

Biosafety &&Human Practice

Experiment

Technical BackgroundTechnical Background

Create a star!

Main GoalMain Goal : Engineering bacteria colony to automatically break their own symmetry

: Engineering bacteria colony to automatically break their own symmetry

Whole System

AHL transduction

Light sensor

Toggle switch

Toggle switch

Antisense FtsZ repress division

Antisense FtsZ repress division

FtsZ mRNA

FtsZ asRNA

FtsZ geneFtsZ protein

silencing

transcription

translate

Tby gene transcription

Lux I gene Lux I protein

SAM

AHL

luxR gene express luxR protein

combine

Luxpr promoterpromote

pcI promoter

Lac promoter

lacI promoter

lacI gene

cI gene cI protein

lacI proteinpromote

promoteinhibit

inhibit

IPTG

ultraviolet rays

promote

Light sensor promoter

cI gene cI proteinpromote

Light receiverexpress

Red light

OmpC OmpC

express

express

express

express

Light Sensor

Red Light11

Blue Light22

Red Light

Light Sensor

Cph8Cph8

RegulatoryBLp

ycgZ ycgF ycgE

regulated by the ycgF/E system

Blue Light

Conformation

Binding&&Repress Expression

Blue Light

Light Sensor

Toggle Switch

Toggle Switch

lacI+pL LuxR LuxIGenerating More AHL

Activating

AHL-Based Quorum Sensing

Antisense RNA

Antisense RNA

Verification for Antisense RNA

IPTG=0

After 10 hours in 37°C environment

IPTG=100 IPTG=200

Design Scheme

Related Kits:

j5526=r11+b34+mRFP+tt i13507=b34+mRFP+tt i13521=tetR+b34+mRFP+tt j04450=lac+b34+mRFP+tt

r0011=lacipl r0051=pci r0040=ptetr r0063=luxpl(ahl--) r0062=luxpr(ahl++) r1051=r0051=pcI

i732820=b34+LacI+tt j23039=tetR+b34+luxI+tt i13973=pcI+b34+yfp+tt k098991=pcI+b34+gfp+tt p0451=b34+cI+tt k092400=b34+luxi+tt i13202=lacipl+b34+luxI+tt

t9002=tetr+b34+luxr+tt+luxpr+b32+gfp+tt f2620=ptetr+b34+luxr+tt+luxpr

k145270=pro+b34+luxr+tt j37033=b34+luxr k091204=pro+b34+luxr+tt i0462=b34+luxr+tt k116640=ptetR+b32+luxr+tt j37015=ptetR+b34+luxr+tt+luxpr+b34+luxI+b34+gfp+tt

K088006=b33+lacI+lac+b34+cI+b32+gfp+tt+pcI+b34+rfp+tt

I763026=ptetR+b34+lacI+tt I763035=ptetr+b34+lacI+b34+gfp+tt p0151=b31+cI+tt p0440=b34+tetr+tt

k145205=ptetr+b32+gfp+tt K082034=lac+b34+gfp+tt

The Simulation Experiment

The Micro-Model

-Based on Cellular Automaton

The Macro-Model

-Based on Improved Cellular

Automaton

Modeling

Approximate Bayesian Computation （ ABC ） for parameter inference in gene expression model

Modeling

1. Approximate Bayesian computation (ABC)

ABC rejection sampler

ABC MCMC

2. Gene expression model

Modeling

Results

Results

Verification for Antisense RNA

IPTG=0

After 10 hours in 37°C environment

IPTG=100 IPTG=200

others

Potential applications

Biosensor

Cell Printing

Signal Transnduction

Potential applications

Acknowledgements

We would like to thank :Academy of Biomedical Engineering for all these support;

数学模型数学模型

文档文档

反义反义

光敏启动光敏启动

开关开关

AHLAHLOthers

③

②

①

该项目大部分由我们自主完成（除了非天然序列发夹结构外包合成），技术上不依赖外援。

项目总结 由于缺乏经验，该项目实现效果不如我们预期，但已部分实现。

下一步我们将完成元件的标准化和功能验证，以及各种材料的整理和项目展示工作。