Team:Northwestern

From 2013.igem.org

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<p id = "project-description-title"> <font size=1>N</font> <font size=2>N</font> <font size=3>N</font> <font size=4>N</font> <font size=5>N</font> NUtralize! </p>  
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    This year the Northwestern iGEM team has decided to focus on oral health care as the theme of our project. It has been well characterized that lactic acid secreted by natural bacteria in the human mouth decreases pH, leading to cavity formation. The enamel demineralization threshold is known to be around pH 5.5. Previous research has found that the genes asr and gadA are highly expressed near pH 5.5 and below. Our project aims to take a two-pronged approach at combating the natural lactic acid secretion. We plan to create a promoter that is always on at a basal level as well as one that is induced to express at a high level as a result of low pH. This project will lay the foundation for a significant amount of research as the dual-state promoter that we develop could then be incorporated with a gene related to lactic acid catabolism or neutralization.  
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    Poor oral health (tooth decay and infection) can lead to many painful symptoms and complicated treatments. As mouth bacteria are a primary culprit in tooth decay, as lactic acid secreted by natural bacteria in the human mouth decreases pH, leading to cavity formation. The enamel demineralization threshold is known to be around pH 5.5. The overall vision is to prevent mouth pH from falling below 5.5 by engineering bacteria to sense pH and respond by increasing alkalinity. Our summer project aimed at creating genetic pH sensing elements that would work in the mouth.  Previous research has found that the genes asr and gadA are highly expressed when  pH drops below 5.5. Our project laid the foundation for a dual-state promoter consisting of two promoters in series with a constitutive promoter on at a (tunable) basal level and a downstream pH inducible promoter. In the process, we have characterized a new pH responsive promoter, asr, as well as the individual gadA and constitutive promoters.  Ongoing work will evaluate how promoters in series affect each other and characterize the dual state phenomenon.  This project lays the groundwork for sensing pH drops and could induce enzymes that catalyze reactions to increase the pH.  
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Revision as of 02:38, 27 September 2013



NUtralize!

Poor oral health (tooth decay and infection) can lead to many painful symptoms and complicated treatments. As mouth bacteria are a primary culprit in tooth decay, as lactic acid secreted by natural bacteria in the human mouth decreases pH, leading to cavity formation. The enamel demineralization threshold is known to be around pH 5.5. The overall vision is to prevent mouth pH from falling below 5.5 by engineering bacteria to sense pH and respond by increasing alkalinity. Our summer project aimed at creating genetic pH sensing elements that would work in the mouth. Previous research has found that the genes asr and gadA are highly expressed when pH drops below 5.5. Our project laid the foundation for a dual-state promoter consisting of two promoters in series with a constitutive promoter on at a (tunable) basal level and a downstream pH inducible promoter. In the process, we have characterized a new pH responsive promoter, asr, as well as the individual gadA and constitutive promoters. Ongoing work will evaluate how promoters in series affect each other and characterize the dual state phenomenon. This project lays the groundwork for sensing pH drops and could induce enzymes that catalyze reactions to increase the pH.

Summary of Problem

Following meals, the pH within the mouth drops substantially which leads to cavities.

pH

The first step in combating cavities is to create a bacteria that can detect pH.

Dual State Promoter

In order for a quick response from basal level, we designed the dual state promoter.