iGEM 2020 — Team UFlorida

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# Team:UFlorida/Notebook

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TEAM

[Team](https://2020.igem.org/Team%3AUFlorida/Team)
[Attributions](https://2020.igem.org/Team%3AUFlorida/Attributions)
[Collaborations](https://2020.igem.org/Team%3AUFlorida/Collaborations)

PROJECT

[Description](https://2020.igem.org/Team%3AUFlorida/Description)
[Contribution](https://2020.igem.org/Team%3AUFlorida/Contribution)
[Implementation](https://2020.igem.org/Team%3AUFlorida/Implementation)
[Engineering Success](https://2020.igem.org/Team%3AUFlorida/Engineering)

[PARTS](https://2020.igem.org/Team%3AUFlorida/Parts)
[HUMAN PRACTICES](https://2020.igem.org/Team%3AUFlorida/Human_Practices)

AWARDS

[Education](https://2020.igem.org/Team%3AUFlorida/Education)
[Model](https://2020.igem.org/Team%3AUFlorida/Model)

[JUDGING FORM ⇗](https://igem.org/2020_Judging_Form?team=UFlorida)

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# Notebook

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# Team:UFlorida/Engineering

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TEAM

[Team](https://2020.igem.org/Team%3AUFlorida/Team)
[Attributions](https://2020.igem.org/Team%3AUFlorida/Attributions)
[Collaborations](https://2020.igem.org/Team%3AUFlorida/Collaborations)

PROJECT

[Description](https://2020.igem.org/Team%3AUFlorida/Description)
[Contribution](https://2020.igem.org/Team%3AUFlorida/Contribution)
[Implementation](https://2020.igem.org/Team%3AUFlorida/Implementation)
[Engineering Success](https://2020.igem.org/Team%3AUFlorida/Engineering)

[PARTS](https://2020.igem.org/Team%3AUFlorida/Parts)
[HUMAN PRACTICES](https://2020.igem.org/Team%3AUFlorida/Human_Practices)

AWARDS

[Education](https://2020.igem.org/Team%3AUFlorida/Education)
[Model](https://2020.igem.org/Team%3AUFlorida/Model)

[JUDGING FORM ⇗](https://igem.org/2020_Judging_Form?team=UFlorida)

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##### Research → Imagine → Design → Build → Test → Learn → Improve → Research...

### RESEARCH

Harmful algal blooms, such as “red tide” caused by Karena Brevis, have a direct negative environmental and economic impact in Florida. Wide-scale fish kills and the release of neurotoxins harm wildlife that inhabit the coastal waters and people who depend on the environment to run their businesses. Eutrophication also contributes to a negative environmental impact in the state of Florida, as it causes excess algal growth. In turn, the excess algae deplete oxygen and essential nutrients for other organisms in the habitat, leading to the loss of biodiversity. A main contributor to eutrophication and harmful algal blooms are fertilizer runoffs from the land, with a major ingredient in fertilizer being phosphorous.

### IMAGINE

Team Florida strives to protect our waterways and wetland ecosystems from this devastation. The first step in solving the problem is detection, so Team UFlorida has taken on the challenge of creating a biosensor that can detect above-safe levels of phosphorus in samples of water. This biosensor would allow ecologists to make informed decisions about the impacted waterways. The biosensor would be integrated into E.coli and would serve to quantify the amount of phosphorus in a given body of water.

### DESIGN

Team Florida saw the potential to engineer a naturally occurring system to produce a desired response. This is where Team Florida built upon their previous project to couple the PhoB-PhoR system with SCRIBE and a genetic inverter that would allow this system to respond at phosphorous concentrations above 300uM instead of below these conditions.

By adding our own genetic inverter, we have genetically engineered the PhoB-PhoR system to enable it to respond to high phosphorus levels. The PhoB-PhoR network has a histidine kinase sensor (PhoR) and a response regulator protein (PhoB). When E.coli cells are low in phosphate, these organisms activate gene expression of genes like PhoR which serve as a sensor protein that when stimulated by the lack of phosphate, catalyzes the activation of response regulator PhoB. As a positive response regulator, PhoB activates the gene expression of itself, alongside PhoR and other genes essential for the intake of (Pi) into E.coli. PhoB then goes on to directly active promoter PhoA.

Tetr encodes for a repressor protein that comes directly after the pPhoA promoter. Then, there is a pTet promoter after the Tetr gene. In low phosphate conditions, PhoB binds to activate pPhoA leading to the transcription of the Tetr repressor gene. The Tetr repressor gene is then translated into the Tetr protein, which binds and represses the pTet promoter; therefore, SCRIBE is not transcribed.

Because pTet is a constitutive promoter, when the biosensor is placed in high phosphate conditions, PhoBR is turned off and the pTet promoter remains active due to the absence of Tetr. Thus, the SCRIBE system is under control of the pTet promoter. SCRIBE is transcribed and a mutation results in the bacterial chromosome.

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# Team:UFlorida/Experiments

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TEAM

[Team](https://2020.igem.org/Team%3AUFlorida/Team)
[Attributions](https://2020.igem.org/Team%3AUFlorida/Attributions)
[Collaborations](https://2020.igem.org/Team%3AUFlorida/Collaborations)

PROJECT

[Description](https://2020.igem.org/Team%3AUFlorida/Description)
[Contribution](https://2020.igem.org/Team%3AUFlorida/Contribution)
[Implementation](https://2020.igem.org/Team%3AUFlorida/Implementation)
[Engineering Success](https://2020.igem.org/Team%3AUFlorida/Engineering)

[PARTS](https://2020.igem.org/Team%3AUFlorida/Parts)
[HUMAN PRACTICES](https://2020.igem.org/Team%3AUFlorida/Human_Practices)

AWARDS

[Education](https://2020.igem.org/Team%3AUFlorida/Education)
[Model](https://2020.igem.org/Team%3AUFlorida/Model)

[JUDGING FORM ⇗](https://igem.org/2020_Judging_Form?team=UFlorida)

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# TITLE

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# Team:UFlorida/Results

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[HOME](https://2020.igem.org/Team%3AUFlorida)

TEAM

[Team](https://2020.igem.org/Team%3AUFlorida/Team)
[Attributions](https://2020.igem.org/Team%3AUFlorida/Attributions)
[Collaborations](https://2020.igem.org/Team%3AUFlorida/Collaborations)

PROJECT

[Description](https://2020.igem.org/Team%3AUFlorida/Description)
[Contribution](https://2020.igem.org/Team%3AUFlorida/Contribution)
[Implementation](https://2020.igem.org/Team%3AUFlorida/Implementation)
[Engineering Success](https://2020.igem.org/Team%3AUFlorida/Engineering)

[PARTS](https://2020.igem.org/Team%3AUFlorida/Parts)
[HUMAN PRACTICES](https://2020.igem.org/Team%3AUFlorida/Human_Practices)

AWARDS

[Education](https://2020.igem.org/Team%3AUFlorida/Education)
[Model](https://2020.igem.org/Team%3AUFlorida/Model)

[JUDGING FORM ⇗](https://igem.org/2020_Judging_Form?team=UFlorida)

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