Difference between revisions of "Team:Kingsborough NY/LacZ"
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<h1>Figure 1: LacZ expression of K2268004 grown in the Dark</h1> | <h1>Figure 1: LacZ expression of K2268004 grown in the Dark</h1> | ||
<p>[[File:Kbcc_3.211111.gif|700px|center]]</p> | <p>[[File:Kbcc_3.211111.gif|700px|center]]</p> | ||
| − | <p>( | + | <p>(Figure 1)Shows the plating of K2268004 grown in the dark where the blue indicator shows the direction of plating. The center had the highest expression of LacZ suggesting that rate of cell growth is the highest at the center and rate of cell growth towards the edge of the plate is lower. The cell was induced with 1mM of IPTG in 20 ɥg/ɥl of X-gal. |
</p> | </p> | ||
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<h1>Figure 2: Approach for ImageJ Analysis (Dark)</h1> | <h1>Figure 2: Approach for ImageJ Analysis (Dark)</h1> | ||
<p>[[File:Kbcc_mark_3.2dark.jpg|700px|center]]</p> | <p>[[File:Kbcc_mark_3.2dark.jpg|700px|center]]</p> | ||
| − | <p>( | + | <p>(Figure 2)Shows the ImageJ analysis for the average expression level. Each “cell” is a ratio of blue to gray and expression level was found by taking the ratio of the background to the cell. |
</p> | </p> | ||
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<h1>Figure 3: LacZ expression of K2268003 grown in the Light</h1> | <h1>Figure 3: LacZ expression of K2268003 grown in the Light</h1> | ||
<p>[[File:Kbcc_light3.2.gif|750px|center]]</p> | <p>[[File:Kbcc_light3.2.gif|750px|center]]</p> | ||
| − | <p>( | + | <p>(Figure 3)Shows the plating of K2268003 grown in the light where the blue indicator shows the direction of plating. The center had the highest expression of LacZ suggesting that rate of cell growth is the highest at the center and rate of cell growth towards the edge of the plate is lower. The cell was induced with 1mM of IPTG in 20 ɥg/ɥl of X-gal. |
</p> | </p> | ||
<h1>Figure 4: Approach for ImageJ Analysis (Light)</h1> | <h1>Figure 4: Approach for ImageJ Analysis (Light)</h1> | ||
<p>[[File:Kbcc_mark_3.2light.jpg|750px|center]]</p> | <p>[[File:Kbcc_mark_3.2light.jpg|750px|center]]</p> | ||
| − | <p>( | + | <p>(Figure 4)Shows the ImageJ analysis for the average expression level. Each “cell” is a ratio of blue to gray and expression level was found by taking the ratio of the background to the cell. |
</p> | </p> | ||
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<h1>Figure 6: Optimizing X-gal concentrations</h1> | <h1>Figure 6: Optimizing X-gal concentrations</h1> | ||
<p>[[File:Kbcc-20-2017.jpg|500px|left]][[File:Kbcc-40-2017.jpg|500px|right]]</p> | <p>[[File:Kbcc-20-2017.jpg|500px|left]][[File:Kbcc-40-2017.jpg|500px|right]]</p> | ||
| − | <p>( | + | <br clear=all> |
| + | <p>(Figure 6) shows that increasing x-gal from 20 ɥg/ɥl to 40 ɥg/ɥl showed a noticeable increase in LacZ expression for K2268004. Increased expression level shows that LacI inhibition is not strong and that LacZ expression is coming from the plasmid backbone (pSB1C3) or the bacteria itself.</p> | ||
<br clear=all> | <br clear=all> | ||
Revision as of 01:33, 2 November 2017
This page demonstrates the activity and function of our control constructs, K2268003 and K2268004, by measuring LacZ activity.
Contents
- 1 Figure 1: LacZ expression of K2268004 grown in the Dark
- 2 Figure 2: Approach for ImageJ Analysis (Dark)
- 3 Figure 3: LacZ expression of K2268003 grown in the Light
- 4 Figure 4: Approach for ImageJ Analysis (Light)
- 5 Figure 5: Quantified Results comparing Light and Dark conditions
- 6 Figure 6: Optimizing X-gal concentrations
Figure 1: LacZ expression of K2268004 grown in the Dark
(Figure 1)Shows the plating of K2268004 grown in the dark where the blue indicator shows the direction of plating. The center had the highest expression of LacZ suggesting that rate of cell growth is the highest at the center and rate of cell growth towards the edge of the plate is lower. The cell was induced with 1mM of IPTG in 20 ɥg/ɥl of X-gal.
Figure 2: Approach for ImageJ Analysis (Dark)
(Figure 2)Shows the ImageJ analysis for the average expression level. Each “cell” is a ratio of blue to gray and expression level was found by taking the ratio of the background to the cell.
Figure 3: LacZ expression of K2268003 grown in the Light
(Figure 3)Shows the plating of K2268003 grown in the light where the blue indicator shows the direction of plating. The center had the highest expression of LacZ suggesting that rate of cell growth is the highest at the center and rate of cell growth towards the edge of the plate is lower. The cell was induced with 1mM of IPTG in 20 ɥg/ɥl of X-gal.
Figure 4: Approach for ImageJ Analysis (Light)
(Figure 4)Shows the ImageJ analysis for the average expression level. Each “cell” is a ratio of blue to gray and expression level was found by taking the ratio of the background to the cell.
Figure 5: Quantified Results comparing Light and Dark conditions
(Figure5)Shows that there is a significant difference in expression level between K2268005 grown in light and K2268005 grown in dark with significantly higher expression in K2268005 grown in light, p<.001. Data analysis was done by comparing the mean expression level of figure 2 and figure 4 with GraphPad Software.
Figure 6: Optimizing X-gal concentrations
(Figure 6) shows that increasing x-gal from 20 ɥg/ɥl to 40 ɥg/ɥl showed a noticeable increase in LacZ expression for K2268004. Increased expression level shows that LacI inhibition is not strong and that LacZ expression is coming from the plasmid backbone (pSB1C3) or the bacteria itself.
