Team:Ehime-Japan/Modeling
From 2012.igem.org
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+ | <p><font size="5"> | ||
+ | <p>Introduction</p> | ||
+ | <b> | ||
+ | <p><font size="3"> | ||
+ | In our experiment of the E.co-mail, the important thing is to degrade GFP-Lon-tag more rapidly. GFP is usually stable and has a long half–life. So, we thought of how to measure the reaction rate. | ||
+ | <p/> | ||
+ | <p><font size="5"> | ||
+ | <p>Method<p/> | ||
+ | <b><font size="3"> | ||
+ | E.coli (pIJ106b, pJT122, plPCB) is cultivated in TY culture medium for 7 hours. It was precipitated by centrifugation and washed with 1 % NaClaq. | ||
+ | 400 μL of 1% NaCl was added to the E.coli sample. | ||
+ | An aliquot from each sample was plated on agar and cultured overnight. | ||
+ | The solution was exposed to blue LED light, and we took a color picture every 1 hour for 6 hours. | ||
+ | By using the ImageJ software, we measured the intensity of light from GFP. | ||
+ | <b> | ||
+ | <p><font size="5"> | ||
- | + | <p>Result<p/> | |
+ | <b> | ||
+ | <p><font size="3"> | ||
+ | The relative light intensity values are summarized in Table 1. As we can only obtain relative values as intensity of light, we set the value for the 0-hour sample as 1. | ||
+ | <table> | ||
+ | <tr> | ||
+ | <th>TABlE 1 Intensity of light</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td> | ||
+ | <img src="https://static.igem.org/mediawiki/2012/6/69/%E7%84%A1%E9%A1%8Ca.png"width="600"></td> | ||
+ | </table> | ||
+ | |||
+ | <p/> | ||
+ | <img src="https://static.igem.org/mediawiki/2012/a/ac/%E7%84%A1a%E9%A1%8C.png"> | ||
+ | Fig Relation between Intensity of light and time<br> | ||
+ | We did not use date of 5 hour and 6 hour. | ||
+ | |||
+ | <br> | ||
+ | |||
+ | <p><font size="5"><p/> | ||
+ | <p>Discussion<p/> | ||
+ | <b> | ||
+ | <p><font size="3"><p/> | ||
+ | Because there is the propotional relationship between light intensity and GFP concentration, we made a graph of this and took collinear approximation. Besides considering parameter, three is this approximation straight line is primary rate equation. | ||
+ | |||
+ | <b><p> | ||
+ | (d [GFP-Lon tag])/dt = -k1 [GFP-Lon tag][Lon-LVA] | ||
+ | <br> | ||
+ | (d [Lon-ClpXP])/dt = -k2 [Lon – LVA][LVA] +Lon synthesis rate | ||
+ | <p/> | ||
</body> | </body> | ||
+ | |||
</html> | </html> |
Latest revision as of 03:59, 27 September 2012
Introduction
In our experiment of the E.co-mail, the important thing is to degrade GFP-Lon-tag more rapidly. GFP is usually stable and has a long half–life. So, we thought of how to measure the reaction rate.
Method
Result
The relative light intensity values are summarized in Table 1. As we can only obtain relative values as intensity of light, we set the value for the 0-hour sample as 1.
Discussion
Because there is the propotional relationship between light intensity and GFP concentration, we made a graph of this and took collinear approximation. Besides considering parameter, three is this approximation straight line is primary rate equation.
(d [GFP-Lon tag])/dt = -k1 [GFP-Lon tag][Lon-LVA]
Fig Relation between Intensity of light and time
TABlE 1 Intensity of light
We did not use date of 5 hour and 6 hour.
(d [Lon-ClpXP])/dt = -k2 [Lon – LVA][LVA] +Lon synthesis rate