Difference between revisions of "Part:BBa K861169:Experience"
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===Applications of BBa_K861169=== | ===Applications of BBa_K861169=== | ||
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| + | <html> | ||
| + | <h2>Device Design</h2> | ||
| + | <p algin="justify"> | ||
In this design, we used promoter PcstA and an intermediate regulator cI, as PcstA is activated by CRP in low glucose concentration, whereas we need a device boosted by high concentration of glucose. | In this design, we used promoter PcstA and an intermediate regulator cI, as PcstA is activated by CRP in low glucose concentration, whereas we need a device boosted by high concentration of glucose. | ||
| − | As you can see, in this pathway, protein cI will be expressed when CRP is activated under low glucose concentration.The large amount of cI repressed the promoter heavily. On the contrary, when glucose concentration is high, the promoter is derepressed. | + | As you can see, in this pathway, protein cI will be expressed when CRP is activated under low glucose concentration.The large amount of cI repressed the promoter R0051 heavily. On the contrary, when glucose concentration is high, the promoter is derepressed.</p> |
| − | We made RFP as reporter and certify that the design works well via two different methods. | + | <p algin="justify"> |
| + | We made RFP as reporter and certify that the design works well via two different methods.</p> | ||
| − | + | <h2>Fluorescence measurement</h2> | |
| + | <p algin="justify"> | ||
In the first method, Fluorescence intensity of cell culture was directly read from a plate reader. And all data is normalized by OD600. | In the first method, Fluorescence intensity of cell culture was directly read from a plate reader. And all data is normalized by OD600. | ||
| − | In this figure, the fluorescence of RFP generator promoted by PcstA decreased when concentration of glucose rose, meanwhile, the fluorescence of the indirect regulatory device increased with the glucose concentration, indicating that the device works as expected. | + | </p> |
| − | + | ||
| − | The second method is practiced for single cells. The fluorescence pictures of different glucose concentration were captured by fluorescence microscope. Also, a program named FANCY was designed to recognize single cell and calculate the fluorescence strength in the images. As the result shows, it conformed well to that of the first method. | + | <p align="justify"><img name="" src="https://static.igem.org/mediawiki/igem.org/9/9c/Indirect_Fig_1.png" width="506" height="409" alt=""></p> |
| − | + | ||
| + | <p algin="justify"> | ||
| + | In this figure, the fluorescence of RFP generator promoted by PcstA decreased when concentration of glucose rose, meanwhile, the fluorescence of the indirect regulatory device increased with the glucose concentration, indicating that the device works as expected.</p> | ||
| + | <p align="justify"><img name="" src="https://static.igem.org/mediawiki/igem.org/d/dc/Indirect_Fig_2.png" width="500" height="220" alt=""></p> | ||
| + | <p align="justify"><img name="" src="https://static.igem.org/mediawiki/igem.org/2/21/Indirect_Fig_3.png" width="506" height="189" alt=""></p> | ||
| + | <p align="justify"><img name="" src="https://static.igem.org/mediawiki/igem.org/c/c3/Indirect_Fig_4.png" width="500" height="226" alt=""></p> | ||
| + | <p algin="justify"> | ||
| + | The second method is practiced for single cells. The fluorescence pictures of different glucose concentration were captured by fluorescence microscope. Also, a program named <a href="http://2012.igem.org/Team:WHU-China/Modeling?catalog=3">FANCY</a> was designed to recognize single cell and calculate the fluorescence strength in the images. As the result shows, it conformed well to that of the first method.</p> | ||
| + | <p algin="justify"> | ||
All these results indicated that the device we designed can meet the demand, promoters activated by glucose. | All these results indicated that the device we designed can meet the demand, promoters activated by glucose. | ||
| + | </p> | ||
| + | </html> | ||
===User Reviews=== | ===User Reviews=== | ||
Latest revision as of 03:35, 27 September 2012
This experience page is provided so that any user may enter their experience using this part.
Please enter
how you used this part and how it worked out.
Applications of BBa_K861169
Device Design
In this design, we used promoter PcstA and an intermediate regulator cI, as PcstA is activated by CRP in low glucose concentration, whereas we need a device boosted by high concentration of glucose. As you can see, in this pathway, protein cI will be expressed when CRP is activated under low glucose concentration.The large amount of cI repressed the promoter R0051 heavily. On the contrary, when glucose concentration is high, the promoter is derepressed.
We made RFP as reporter and certify that the design works well via two different methods.
Fluorescence measurement
In the first method, Fluorescence intensity of cell culture was directly read from a plate reader. And all data is normalized by OD600.
In this figure, the fluorescence of RFP generator promoted by PcstA decreased when concentration of glucose rose, meanwhile, the fluorescence of the indirect regulatory device increased with the glucose concentration, indicating that the device works as expected.
The second method is practiced for single cells. The fluorescence pictures of different glucose concentration were captured by fluorescence microscope. Also, a program named FANCY was designed to recognize single cell and calculate the fluorescence strength in the images. As the result shows, it conformed well to that of the first method.
All these results indicated that the device we designed can meet the demand, promoters activated by glucose.
User Reviews
UNIQ2b024eee857ba5c3-partinfo-00000001-QINU UNIQ2b024eee857ba5c3-partinfo-00000002-QINU