Difference between revisions of "Team:HKUST-Rice/Nitrate Sensor PyeaR"
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<h1>Experiment that we did</h1> | <h1>Experiment that we did</h1> | ||
| − | <p> | + | <p>We have done two sets of characterization on pSB1C3-BBa_K381001 (BCCS-Britstol 2010), one using LB medium and the other in M9 minimal medium. We did quantitative characterization on the promoter by measuring the fluorescence signal intensity using a plate reader.</P> |
| − | + | <p><b><u>pSB1C3-BBa_K381001 characterization</u></b><br><b>Medium: LB</b><br><br><b>Responsive range of promoter characterization in LB</b><br> | |
| − | + | The concentration of the characterization of PyeaR promoter was from 0 to 50mM Nitrate, with an intervals of 10mM. | |
| − | + | Potassium Nitrate was being used as the source of nitrate in our experiments. | |
| + | 1M KNO3 stock solution was first made and added in the following ratios to produce different concentrations of medium. | ||
| + | E. coli strain DH10B was used in the characterization of the promoter.</p> | ||
| + | </p> | ||
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<img src="https://static.igem.org/mediawiki/2015/b/be/HKUST-Rice15_Resultsbutton.png" alt="image caption"> | <img src="https://static.igem.org/mediawiki/2015/b/be/HKUST-Rice15_Resultsbutton.png" alt="image caption"> | ||
Revision as of 02:40, 29 August 2015
Nitrate Sensor
Nitrate as a Macro-nutrient
#
Nitrate sensor Design
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, some will relieve the repression from the nar system and others 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. As a result, the reporter signal will increase with increasing nitrate concentration.
Experiment that we did
We have done two sets of characterization on pSB1C3-BBa_K381001 (BCCS-Britstol 2010), one using LB medium and the other in M9 minimal medium. We did quantitative characterization on the promoter by measuring the fluorescence signal intensity using a plate reader.
pSB1C3-BBa_K381001 characterization
Medium: LB
Responsive range of promoter characterization in LB
The concentration of the characterization of PyeaR promoter was from 0 to 50mM Nitrate, with an intervals of 10mM.
Potassium Nitrate was being used as the source of nitrate in our experiments.
1M KNO3 stock solution was first made and added in the following ratios to produce different concentrations of medium.
E. coli strain DH10B was used in the characterization of the promoter.
Result obtained
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Lorem ipsum dolor sit amet, pro aeque temporibus eu, eum qualisque assueverit te. Ad est admodum epicuri suscipit, te alterum aliquando adversarium usu, pro ex omnesque luptatum comprehensam. In vix alia percipit gloriatur, no ferri lorem aliquando cum. Fugit concludaturque sed ne, ea sumo dico adolescens eos, quo eu pertinax expetendis. An his omnes instructior, vide possim eam id. Te cum enim sale offendit, vocent copiosae luptatum ut per. Eam in alienum accusamus, et probo reque vix. Vivendum necessitatibus qui ad, no vis enim veniam perpetua. Eu pri habemus senserit, dicit tation expetenda usu et. Sea eu dolor deserunt dissentias, sed an oportere moderatius assueverit. Usu te tation gloriatur, vidit tollit utinam mea id.