Difference between revisions of "Team:HKUST-Rice/Phosphate Sensor"
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| − | <div id= "page_title"><h1>Phosphate Sensor - | + | <div id= "page_title"><h1>Phosphate Sensor - <i>phoAp</i> , <i>phoBRp</i></h1> |
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<h1>Introduction</h1> | <h1>Introduction</h1> | ||
| − | <p>Phosphorus plays an essential role in plant growth. It | + | <p>Phosphorus plays an essential role in plant growth. It associates with various growth factors for root development and seed production, etc. Deficiency in Phosphorus lead to stunted growth, yet symptoms are not obvious. Therefore, it is important to monitor the level of both the organic and inorganic phosphorus level in soil for healthy plant growth. |
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| − | <p><i>Escherichia coli</i> detects inorganic phosphate (P(i)) from the environment by the PhoR/PhoB | + | <p><i>Escherichia coli</i> detects inorganic phosphate (P(i)) from the environment by the PhoR/PhoB two-component system (Hsieh & Wanner, 2010). As illustrated in Figure.1, <i>phoAp</i> and <i>phoBRp</i> is cross-regulated by PhoB and PhoR. The sensory histidine kinase PhoR can behaves as an activator or inactivator for PhoB depending on different states (inhibition state, activation state, deactivation state). When phosphate is limited, PhoR act as a phospho-donor for the autophosphorylation of PhoB. The phosphorylated PhoB will directly activate <i>phoAp</i> and <i>phoBRp</i>. In contrast, when there is phosphate, PhoR interferes with phosphorylation of PhoB which in turn inactivates the <i>phoAp</i> and <i>phoBR</i>. |
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| − | <br><br>With the phosphate (Pho) regulon from <i>E. coli</i>,it can be utilized for detecting phosphate level. | + | <br><br>With the phosphate (Pho) regulon from <i>E. coli</i>,it can be utilized for detecting phosphate level. <i>phoAp</i> and <i>phoBR</i> from <i>E. coli</i> strain DH10B were cloned and ligated with the GFP generator (<a href="http://parts.igem.org/Part:BBa_I13504"target="_blank">pSB1C3-BBa_I13504</a>) respectively. Under high phosphate concentrations, repression on the green fluorescence intensity is expected; while under low phosphate concentrations, expression on green fluorescence is expected.</p> |
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<h1>Results obtained</h1> | <h1>Results obtained</h1> | ||
| − | <p>Characterization on the contructs (<a>BBa_K1682013</a>) using <a>M9 minimal medium (without phosphate)</a> were performed. Quantitative | + | <p>Characterization on the contructs (<a href="http://parts.igem.org/Part:BBa_K1682013"target="_blank">BBa_K1682013</a>) using <a>M9 minimal medium (without phosphate)</a> were performed. Quantitative characterization on the promoters were done by measuring the fluorescence signal intensity using an EnVision multilabel reader.<br><br>The results were obtained by combining the 3 characterization results together.</p> |
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Revision as of 13:37, 3 September 2015
Phosphate Sensor - phoAp , phoBRp
Introduction
Phosphorus plays an essential role in plant growth. It associates with various growth factors for root development and seed production, etc. Deficiency in Phosphorus lead to stunted growth, yet symptoms are not obvious. Therefore, it is important to monitor the level of both the organic and inorganic phosphorus level in soil for healthy plant growth.
Phosphate sensor Design
Escherichia coli detects inorganic phosphate (P(i)) from the environment by the PhoR/PhoB two-component system (Hsieh & Wanner, 2010). As illustrated in Figure.1, phoAp and phoBRp is cross-regulated by PhoB and PhoR. The sensory histidine kinase PhoR can behaves as an activator or inactivator for PhoB depending on different states (inhibition state, activation state, deactivation state). When phosphate is limited, PhoR act as a phospho-donor for the autophosphorylation of PhoB. The phosphorylated PhoB will directly activate phoAp and phoBRp. In contrast, when there is phosphate, PhoR interferes with phosphorylation of PhoB which in turn inactivates the phoAp and phoBR.
With the phosphate (Pho) regulon from E. coli,it can be utilized for detecting phosphate level. phoAp and phoBR from E. coli strain DH10B were cloned and ligated with the GFP generator (pSB1C3-BBa_I13504) respectively. Under high phosphate concentrations, repression on the green fluorescence intensity is expected; while under low phosphate concentrations, expression on green fluorescence is expected.
Results obtained
Characterization on the contructs (BBa_K1682013) using M9 minimal medium (without phosphate) were performed. Quantitative characterization on the promoters were done by measuring the fluorescence signal intensity using an EnVision multilabel reader.
The results were obtained by combining the 3 characterization results together.
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Reference
Hsieh, Y. J., & Wanner, B. L. (2010). Global regulation by the seven-component P i signaling system. Current opinion in microbiology, 13(2), 198-203.