"
Team:PennState/diauxie/progress
From 2008.igem.org
| Line 185: | Line 185: | ||
<h6>Test Construct</h6> | <h6>Test Construct</h6> | ||
<img src="https://static.igem.org/mediawiki/2008/1/1b/Test_construct.JPG" alt="[Test Consturct]" title="" style="width: 100%; border: solid 1px #000" /> | <img src="https://static.igem.org/mediawiki/2008/1/1b/Test_construct.JPG" alt="[Test Consturct]" title="" style="width: 100%; border: solid 1px #000" /> | ||
| - | |||
| - | |||
| - | |||
<p>Tests were then run to compare the levels of induction with various mixes of xylose, glucose and xylose+glucose. The goal was to obtain a noticeably higher level of induction with the xylose/glucose mixture when compared to the wild-type construct.</p> | <p>Tests were then run to compare the levels of induction with various mixes of xylose, glucose and xylose+glucose. The goal was to obtain a noticeably higher level of induction with the xylose/glucose mixture when compared to the wild-type construct.</p> | ||
| + | </td> | ||
<table> <!-- this table separates content into column-like quadrants --> | <table> <!-- this table separates content into column-like quadrants --> | ||
Latest revision as of 02:19, 30 October 2008
| Home | The Team | The Project | Parts | Notebook |
Diauxie EliminationNHR Biosensors
|
Progress & Results
Each test construct (promoter + GFP) was cloned into the pSB1A2 plasmid and transformed into several E. coli strains: DH5α, W3110 ∆xylB-G, and W3110 ∆xylB-R. Preliminary induction studies were run to find the optimal induction time and to analyze the linear range for OD versus fluorescence. Test Construct![[Test Consturct]](https://static.igem.org/mediawiki/2008/1/1b/Test_construct.JPG)
Tests were then run to compare the levels of induction with various mixes of xylose, glucose and xylose+glucose. The goal was to obtain a noticeably higher level of induction with the xylose/glucose mixture when compared to the wild-type construct. |
 DH5a fluoresence |
 W3110 fluoresence |
These graphs show the normalized fluoresence strength for PN, P1, and P3 xylose promoters induced with xylose, glucose, and a mixture. The W3110 cells have xylE and xylG knocked out while DH5α still contain the natural xylose transport and metabolim. This data shows that there is little effect on the fluorescence intensity using strains with xylE and xylG sequences deleted. Our next step is to transform these promoters into E. coli cells with deleted xylose metabolism and transporters.
 PN induction time |
 P1 induction time |
 P3 induction time |
The cells were induced iduced with sugars and then allowed to grow with sample being removed every half hour. The goal was to find the induction time where fluorescence starts to level off. The intensity has leveled off and begins to drop after 7.5 hours so this was the growth time for all future tests.
 PN linear range |
 P1 linear range |
 P3 linear range |
The three promoters analyzed show linear behavior in the optical density range. The strength of fluorescence did change depending on the promoter. With this information we were able to accurately normalize fluorescence data.
