Team:Northwestern/Project
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Our main project is to simulate mammalian skin using chitin in E. Coli. This process involves four major components: Lawn Formation, Chitin Synthesis, Bacterial Apoptosis, and lac-Operon Signaling. | Our main project is to simulate mammalian skin using chitin in E. Coli. This process involves four major components: Lawn Formation, Chitin Synthesis, Bacterial Apoptosis, and lac-Operon Signaling. | ||
- | A thick bacterial lawn is generated by plating ydgG knock-out mutants | + | A thick bacterial lawn is generated by plating Keio Collection ydgG knock-out mutants in an enriched agar plate. The protein product of ydgG plays an integral role in AI-2 transport. ydgG knock-outs exhibit increased motility which should ultimately allow for a thick and level lawn. |
- | The 3.5kbp Chitin Synthase III gene is extracted from the Saccharomyces Cerevisiae genome. Chitin Synthase III catalyzes the polymerization of chitin | + | The 3.5kbp Chitin Synthase III gene is extracted from the Saccharomyces Cerevisiae genome. Chitin Synthase III catalyzes the polymerization of chitin. |
Apoptosis of cells is achieved by using the bacteriophage lysis cassette built by the Brown '08 iGEM Team. The cassette includes Holin, Endolysin, and Rz Protein genes. These enzymes puncture and degrade the cell membrane which results in lysing of the cell and the release of synthesized chitin. | Apoptosis of cells is achieved by using the bacteriophage lysis cassette built by the Brown '08 iGEM Team. The cassette includes Holin, Endolysin, and Rz Protein genes. These enzymes puncture and degrade the cell membrane which results in lysing of the cell and the release of synthesized chitin. | ||
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Once the lawn is established, an IPTG solution is sprayed over the lawn to induce chitin synthesis (fast response) and apoptosis (slow response). IPTG mimics allolactose which binds to the LacI repressor, causing LacI to detach from DNA and allowing RNA Polymerase to begin transcription. This will allow for a build-up in chitin followed by apoptosis and subsequent release into the extracellular environment. Thus, any abrasions to the chitinous surface can be repaired by spraying IPTG. | Once the lawn is established, an IPTG solution is sprayed over the lawn to induce chitin synthesis (fast response) and apoptosis (slow response). IPTG mimics allolactose which binds to the LacI repressor, causing LacI to detach from DNA and allowing RNA Polymerase to begin transcription. This will allow for a build-up in chitin followed by apoptosis and subsequent release into the extracellular environment. Thus, any abrasions to the chitinous surface can be repaired by spraying IPTG. | ||
- | In mammalian skin, mitosis occurs in the basal layer of the epithelial cells and cells travel outwards towards the surface of the skin as they mature. As the cells move further away from the basal layer, they | + | In mammalian skin, mitosis occurs in the basal layer of the epithelial cells and cells travel outwards towards the surface of the skin as they mature. As the cells move further away from the basal layer, they die and their cytoplasm is released and the cells are filled with keratin, thus forming a continuously regenerating protective layer on the outer-most part of epithelial layer; our project is modeled after this. The bacterial lawn is externally controlled to produce chitin only at the top-most layer and these cells then undergo apoptosis to result in the formation of a chitinous layer at the surface of the lawn. |
Revision as of 02:51, 23 October 2010
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