Difference between revisions of "Team:HKUST-Rice/Description"
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| − | < | + | <div class="one"><font size= "6" color=#000000>Overview</font><br><br> |
| − | <p><font size= "4" color=#000000> Nitrogen (N), Phosphorus (P), and potassium (K) are three macronutrients for plants, and deficiencies in any of these can lead to plant diseases. By creating a biological sensor that can quickly provide soil status to plant owners, we can prevent plant diseases due to the lack of nutrients. In view of this, our team is constructing a biological sensor in <i>E. coli</i>, which can detect NPK levels in the surrounding environment and give responses in the form of colors. In addition, we are characterizing the effects of a dual output system, in contrast to a single output system, in order to anticipate the expression of multiple outputs in a single system.</font></p><br> | + | <p><font size= "4" color=#000000> Nitrogen (N), Phosphorus (P), and potassium (K) are three macronutrients for plants, and deficiencies in any of these can lead to plant diseases. By creating a biological sensor that can quickly provide soil status to plant owners, we can prevent plant diseases due to the lack of nutrients. In view of this, our team is constructing a biological sensor in <i>E. coli</i>, which can detect NPK levels in the surrounding environment and give responses in the form of colors. In addition, we are characterizing the effects of a dual output system, in contrast to a single output system, in order to anticipate the expression of multiple outputs in a single system.</font><br></p></div><br> |
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| + | <p><font size= "3" color=#000000>KdpFABC transporter is a high affinity K+ uptake system (Siebers, A. & Altendorf, K., 1988). The promoter upstream of kdpFABC operon, kdpFp, works under low K+ concentration (Polarek, J. W., et al., 1992; Walderhaug, M. O., et al., 1992). The goal is to characterize kdpFp and build a device which is able to sense different concentration of K+ and express different level of GFP accordingly.</font><br> | ||
| + | <img src= "https://static.igem.org/mediawiki/2015/b/b4/Team-HKUST-Rice-KdpdF_promoter_1.JPG"></p> | ||
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| + | <p><font size= "3" color=#000000><i><br>P<sub>phoA</sub></i> promoter (Hsieh, Y. J., & Wanner, B. L.,2010).is cross-regulated by <i>phoB</i> and <i>phoR</i>, and is usually repressed under high phosphate concentration. <i>phoR</i> behaves as an activator as well as an inactivator for <i>phoB</i>. When phosphate is limited, <i>phoR</i> will phosphorylate <i>phoB</i> and the phosphorylated <i>phoB</i> will directly activates the expression of <i>P<sub>phoA</sub></i> promoter. </font><br> | ||
| + | <img src= "https://static.igem.org/mediawiki/2015/7/7f/Team-HKUST-Rice-Pphoa_characterization.JPG"></p> | ||
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| − | + | <p><font size= "3" color=#000000><i>P<sub>yeaR</sub></i> promoter (Lin, et al, 2007)is normally cross-regulated by the Nar two-component regulatory system (T.Nohno, et,al. , 1989) and <i>nsrR</i>, a regulatory protein. When there is nitrate and nitrite, it will be converted into nitric oxide. The nitric oxide will bind to <i>nsrR</i> and relieve the repression on the <i>P<sub>yeaR</sub></i> promoter. As a result, any genes that are downstream of the <i>P<sub>yeaR</sub></i> promoter will be expressed.</font><br> | |
| − | + | <img src= "https://static.igem.org/mediawiki/2015/0/09/Team-HKUST-Rice-Pyear_characterization.JPG"><p> | |
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| − | + | <p><font size= "3" color=#000000>In order to characterize the output difference between a single expression system and co-expression from a one vector, we will be constructing several inducible systems that give off fluorescence output and compare the dose response of the individual systems in the two scenarios.<br><br>As part of the expression platform, we also aim to construct a 3-input AND logic gate using toehold switches (Green, A. A., et al., 2014) to integrate the input signals detected from the three (N, P and K) promoters.</font><br> | |
| − | + | <img src= "https://static.igem.org/mediawiki/2015/0/08/Team-HKUST-Rice-3inputlogic_gate.JPG"> | |
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<p><font size= "3" color=#000000>Reference: | <p><font size= "3" color=#000000>Reference: | ||
<li>Nohno, T., Noji, S., Taniguchi, S., & Saito, T. (1989). The narX and narL genes encoding the nitrate-sensing regulators of Escherichia coli are homologous to a family of prokaryotic two-component regulatory genes. Nucleic acids research,17(8), 2947-2957.</li><br> | <li>Nohno, T., Noji, S., Taniguchi, S., & Saito, T. (1989). The narX and narL genes encoding the nitrate-sensing regulators of Escherichia coli are homologous to a family of prokaryotic two-component regulatory genes. Nucleic acids research,17(8), 2947-2957.</li><br> | ||
Revision as of 10:18, 27 July 2015
Nitrogen (N), Phosphorus (P), and potassium (K) are three macronutrients for plants, and deficiencies in any of these can lead to plant diseases. By creating a biological sensor that can quickly provide soil status to plant owners, we can prevent plant diseases due to the lack of nutrients. In view of this, our team is constructing a biological sensor in E. coli, which can detect NPK levels in the surrounding environment and give responses in the form of colors. In addition, we are characterizing the effects of a dual output system, in contrast to a single output system, in order to anticipate the expression of multiple outputs in a single system.
KdpFABC transporter is a high affinity K+ uptake system (Siebers, A. & Altendorf, K., 1988). The promoter upstream of kdpFABC operon, kdpFp, works under low K+ concentration (Polarek, J. W., et al., 1992; Walderhaug, M. O., et al., 1992). The goal is to characterize kdpFp and build a device which is able to sense different concentration of K+ and express different level of GFP accordingly.
PphoA promoter (Hsieh, Y. J., & Wanner, B. L.,2010).is cross-regulated by phoB and phoR, and is usually repressed under high phosphate concentration. phoR behaves as an activator as well as an inactivator for phoB. When phosphate is limited, phoR will phosphorylate phoB and the phosphorylated phoB will directly activates the expression of PphoA promoter.
PyeaR promoter (Lin, et al, 2007)is normally cross-regulated by the Nar two-component regulatory system (T.Nohno, et,al. , 1989) and nsrR, a regulatory protein. When there is nitrate and nitrite, it will be converted into nitric oxide. The nitric oxide will bind to nsrR and relieve the repression on the PyeaR promoter. As a result, any genes that are downstream of the PyeaR promoter will be expressed.
In order to characterize the output difference between a single expression system and co-expression from a one vector, we will be constructing several inducible systems that give off fluorescence output and compare the dose response of the individual systems in the two scenarios.
As part of the expression platform, we also aim to construct a 3-input AND logic gate using toehold switches (Green, A. A., et al., 2014) to integrate the input signals detected from the three (N, P and K) promoters.
Reference: