Difference between revisions of "Team:HKUST-Rice/Description"

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<p><font size= "6" color=#6B6B47>Overview</font></p><br>
 
<p><font size= "6" color=#6B6B47>Overview</font></p><br>
 
<p><font size= "4" color=#6B6B47> 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=#6B6B47> 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>
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<p><font size= "4" color=#6B6B47>For nitrate sensing, we decided to use <i>P<sub>yeaR</sub></i> promoter (Lin, et al, 2007). It is normally cross-regulated by the Nar two-component regulatory system (T.Nohno, et,al. , 1989) and <i>nsrR</i>, a regulatory protein that prevents the transcription of a number of operons in <i>E. coli</i>. When there is nitrate and nitrite in the environment, it will enter the cell and then being 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. As the biobrick of <i>P<sub>yeaR</sub></i> promoter with a GFP coding device has already been submitted by the University of Bristol in 2010, we could characterize it and directly apply to our biosensor.</font></p><br>
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<p><font size= "4" color=#6B6B47>For phosphate sensing, we decided to use <i>P<sub>phoA</sub></i> promoter (Hsieh, Y. J., & Wanner, B. L.,2010).. <i>P<sub>phoA</sub></i> promoter is cross-regulated by <i>phoB</i>, a DNA binding response regulator and <i>phoR</i>, a sensory histidine kinase, 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. In contrast, when in high concentration of phosphate, <i>phoR</i> will dephosphorylate <i>phoB</i> and thus, inactivates the expression of <i>P<sub>phoA</sub></i> promoter. As the biobrick of <i>P<sub>phoA</sub></i> promoter with a GFP coding device submitted is not released, we have to clone the promoter from <i>E. coli</i> strain DH10B.</font></p>
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    <td><p><font size= "3" color=#6B6B47>For nitrate sensing, we decided to use <i>P<sub>yeaR</sub></i> promoter (Lin, et al, 2007). It is normally cross-regulated by the Nar two-component regulatory system (T.Nohno, et,al. , 1989) and <i>nsrR</i>, a regulatory protein that prevents the transcription of a number of operons in <i>E. coli</i>. When there is nitrate and nitrite in the environment, it will enter the cell and then being 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. As the biobrick of <i>P<sub>yeaR</sub></i> promoter with a GFP coding device has already been submitted by the University of Bristol in 2010, we could characterize it and directly apply to our biosensor.</font></p><br></td>  
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    <td><p><font size= "3" color=#6B6B47>For phosphate sensing, we decided to use <i>P<sub>phoA</sub></i> promoter (Hsieh, Y. J., & Wanner, B. L.,2010).. <i>P<sub>phoA</sub></i> promoter is cross-regulated by <i>phoB</i>, a DNA binding response regulator and <i>phoR</i>, a sensory histidine kinase, 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. In contrast, when in high concentration of phosphate, <i>phoR</i> will dephosphorylate <i>phoB</i> and thus, inactivates the expression of <i>P<sub>phoA</sub></i> promoter. As the biobrick of <i>P<sub>phoA</sub></i> promoter with a GFP coding device submitted is not released, we have to clone the promoter from <i>E. coli</i> strain DH10B.</font></p></td>
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Revision as of 03:10, 10 July 2015

Overview


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.


For nitrate sensing, we decided to use PyeaR promoter (Lin, et al, 2007). It is normally cross-regulated by the Nar two-component regulatory system (T.Nohno, et,al. , 1989) and nsrR, a regulatory protein that prevents the transcription of a number of operons in E. coli. When there is nitrate and nitrite in the environment, it will enter the cell and then being 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. As the biobrick of PyeaR promoter with a GFP coding device has already been submitted by the University of Bristol in 2010, we could characterize it and directly apply to our biosensor.

For phosphate sensing, we decided to use PphoA promoter (Hsieh, Y. J., & Wanner, B. L.,2010).. PphoA promoter is cross-regulated by phoB, a DNA binding response regulator and phoR, a sensory histidine kinase, 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. In contrast, when in high concentration of phosphate, phoR will dephosphorylate phoB and thus, inactivates the expression of PphoA promoter. As the biobrick of PphoA promoter with a GFP coding device submitted is not released, we have to clone the promoter from E. coli strain DH10B.