"E colock" Chiba 2006

From 2006.igem.org

Contents 1 E colock "self-cycling-E. coli" 2 Member 3 Concept 4 How It Works 5 Techinical Issues 6 Option 7 Work Schedule and Progress

E colock "self-cycling-E. coli"

⇒Go to "E colock" Japanese page??

Member

Chiba 2006 http://chem.tf.chiba-u.jp/igem/ iGEM Chiba

『mu』team

• Shinpei Okuyama
• Akinori Katabami
• Sari Sugaya
• Shinya Matsuno

Concept

By installing circadian rhythms of Cyanobacteria，E. coli starts oscillating.

How It Works

Installing Kai system

Kai system：the simplest and the most well known clock-system with a period of about 24h.

Kai system is driven by phosphorylation cascade composed of three proteins, such as KaiA,KaiB, and KaiC.

The oscillation of Kai system is converted to transcription cycle by Sas A.

This cycle is observable by produced fluorescent proteins.

Techinical Issues

1.Installing Kai Operon

The DNA is installable, whether the Kai system in cyanobacteria is function in E.coli or not?

2.Lack of Synchronization Mechanism

Cyanobacteria use light for “reseting” the cycle, thus its population keeps synchronizing.

However the genes for “reset bottom” has not been identified.

→→①Placing Lac represser under KaiA and KaiBC promoter,Kai oscillation is all stopped. The population is synchronized.

→→②Can we use AI??

Option

Control of cycle duration

There have been KaiA mutants with altered cycle periods. We want to do a similar to create “E clock” s with various cycle periods on demand.

Work Schedule and Progress

08.06 

１.Seek　for　an appropriate　promoter　and　fluoresceint　protein　for this project.←NOW!!
２.Obtain　KaiABC　and　SasA.