"

From 2010.igem.org

Home	Brainstorm	Team	Acknowledgements	Project	Side Project	Parts	Notebook	Calendar	Protocol	Safety	Links	References	Media	Contact

Contents 1 Product Description: Bacterial Skin 2 Side Project: Magnetite Production 3 Northwestern University 4 Sponsors

Product Description: Bacterial Skin

TENSE(?)

Our main project is to simulate mammalian skin using chitin in E Coli. The process was realized by four major components: Lawn Formation, Chitin Synthesis, Bacterial Apoptosis, and lac-Operon Signaling.

A thick bacterial lawn was generated by plating ydgG K.O. mutants (acquired from the Keio Collection) in an enriched agar plate. The protein product of ydgG plays an integral role in AI-2 Transport. Also, the ydgG knock-outs exhibit increased motility. Ultimately, these properties allowed for a thick and level lawn.

Once the lawn was established, a lactose solution was sprayed over the lawn to induce chitin synthesis and apoptosis. Lactose is converted into allolactose which binds to the lacI repressor. Once bound, lacI detaches from DNA to allow RNA Polymerase to begin transcription.

The 3.5kbp ChiA gene was extracted from the Saccharomyces Cerevisiae genome. Chitin Synthase - ChiA protein product - catalyzes the polymerization of chitin by transferring UDP-N-acetyl-D-glucosamine to an N(1,4 N-Acetyl-beta-D-glucosaminyl) to produce N+1(1,4 N-Acetyl-beta-D-glucosaminyl).

Apoptosis of cells was achieved by using the bacteriophage lysis cassette by the Brown '08 iGEM Team. The cassette includes Holin, Endolysin, and Rz Protein genes. Combined, the enzymes puncture and degrade the cell membrane; thus lysing the cell.

In mammalian skin, mitosis occurs in the basal layer of the epithelial cells from which the cells travel. As the cells move further away from the basal layer, they begin to die due to lack of nutrients. They are then filled with keratin and their cytoplasm is released, thus forming a continuously regenerating protective layer on the outer-most part of epithelial layer. Similarly, our bacterial colony will have chitin-filled cells undergo lysis in the top layer of the lawn. The critical difference is that the bacterial colony will not be internally controlled, but this is something that can be remedied, and possibly a goal for future teams.

SIGNIFICANCE:

Chitin itself can be used for:

wound and burn treatment
hemostatic for orthopedic treatment of broken bones
viscoelastic solutions for ophthamology and orthopedic surgery
abdominal adhesions treatment
as antibacterial and antifungal agents and for treatment of mucous membranes
in tumor therapies
in micro surgery, neurosurgery
for treatment of chronic wounds, ulcers and bleeding (chitin powder)    

(http://www.gmp-chitosan.com/en/products-services/chitin.html)

wound healing (CITATION NEEDED)

industrial:

food/pharmaceutical thickener, stabilizer, paper-strengthener

insoluble in water and many organic solvents

etc

agricultural, cosmetics, dietary supplement, water treatment (filter), etc

derivatives such as chitosan have just as many uses 

biodegradable

structural (hard)

Contemporary application:

Internalization and Compartmentalization - and thus sequestering - of particles such as oil

Side Project: Magnetite Production

Our side project is to synthesize superparamagnetic magnetite nanoparticles in E Coli.

Nano-manipulation of magnetite particles, which can lead to a plethora of advances, especially in research technology and biomedical technology.

Understanding and utilization of magnetotaxis and/or magnetoception in bacteria.

Possible production and use of ferrofluid, a paramagnetic or superparamagnetic fluid.

Ability to create uniform nanoparticles will further nanotechnology. (biotech is nanotech that works)

Northwestern University

[http://en.wikipedia.org/wiki/Northwestern_University Northwestern University]

Sponsors