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Revision as of 03:48, 27 October 2010

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Contents 1 Overall project 2 Project Details 2.1 Genetic Circuit 3 Ptet is repressed, and the positive feedback loop is switched on 4 AFP expression ramps up 4.1 The Experiments 4.2 Genetic Circuit Assembly 4.3 Oxygen Sensor 4.4 Results

Overall project

The Penn State Team will be researching and experimenting two different ideas. The first area of study will be creating an oxygen-sensing promoter. This will be done through researching the way that FNR and arcA regulate aerobic/anaerobic respiration in E. coli; engineering applications will be applied to these processes. The second topic we are working on is Bacterial Fireworks, an incorporation of quorum sensing, pigments, and cell lysis. Our goal is to assemble the necessary parts to display the spreading of color as the cells lyse. The team would like to incorporate both aspects of the project to create a bacterial fireworks display that is triggered by a lack of oxygen inside of an agar sphere.

A reliable modular lysis mechanism through quorum sensing could be used in many different fields. As engineered microorganisms are becoming more widely considered for use in the open environment, concerns are growing about possible detrimental effects these microbes could have on ecosystems if they were ever to grow and multiply past expectations. A quorum-based lysis "safety" mechanism would only require one cell to receive a predetermined signal in order to eliminate the entire rogue population, should it escape the area of its intended use.

Project Details

Genetic Circuit

The Penn State iGEM team began work and research in mid May. After learning lab techniques from the grad students, we began researching possible topics. We decided to create a cell lysis device.

Initially, we transformed Lux Promoters, Constitutive Promoters, LuxI + RBS, LuxR + RBS, Lambda Phage lysis devices, Terminators, LacI, LuxR, pLac, CI, RBS, Pigments, FP, LacI + RBS, and CI + RBS.

Lux Promoters	Lux Promoters	Constitutive Promoters	LuxI + RBS	LuxR + RBS	Lambda Phage Lysis Device	Terminator
K091107	R0061	J23108	C0261	J37033	K112022	B0010
K091146	R0063	J23116		S01414	K112808	B1002
K145150	R1062	J23100				B1006

LuxR	LacI	pLac	cI	RBS	Pigment	FP	LacI + RBS	cI + RBS
C0062	C0012	R0010	C0051	B0033	K274100	RFP + RBS	J24679	K081007
				B0034	K274004			P0451

Here is an overview of our genetic circuit. Below, we will take a look at the function of each individual part in detail.

Initially, our genetic circuit cI mechanism

FNR breaks apart in oxygen

Ptet is repressed, and the positive feedback loop is switched on

AFP expression ramps up

The Experiments

Genetic Circuit Assembly

Oxygen Sensor

Results