http://2010.igem.org/wiki/index.php?title=Team:Davidson-MissouriW/MeasuringExpression&feed=atom&action=historyTeam:Davidson-MissouriW/MeasuringExpression - Revision history2024-03-28T14:23:09ZRevision history for this page on the wikiMediaWiki 1.16.5http://2010.igem.org/wiki/index.php?title=Team:Davidson-MissouriW/MeasuringExpression&diff=204300&oldid=prevAnraina at 01:40, 28 October 20102010-10-28T01:40:46Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;The data suggests that the order of genes (TetA and RFP) is important as pLac+RFP+TetA shows more fluorescence in comparison to pLac+TetA+RFP. The trend is reproducible when induced by IPTG (0.5mM) as well. Both cell types, however, fluoresced less than the positive control pLac+RFP. The data also supports our hypothesis that a transcriptional terminator is present in the TetA gene. </p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;The data suggests that the order of genes (TetA and RFP) is important as pLac+RFP+TetA shows more fluorescence in comparison to pLac+TetA+RFP. The trend is reproducible when induced by IPTG (0.5mM) as well. Both cell types, however, fluoresced less than the positive control pLac+RFP. The data also supports our hypothesis that a transcriptional terminator is present in the TetA gene. </p></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><center><img src="https://static.igem.org/mediawiki/2010/<del class="diffchange diffchange-inline">3</del>/<del class="diffchange diffchange-inline">32</del>/<del class="diffchange diffchange-inline">Davidson-MissouriWTPS3</del>.png" <del class="diffchange diffchange-inline">id="fig2" height=600 width=800 </del>></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><center><img src="https://static.igem.org/mediawiki/2010/<ins class="diffchange diffchange-inline">d</ins>/<ins class="diffchange diffchange-inline">de</ins>/<ins class="diffchange diffchange-inline">2ndpic</ins>.png"></center><br></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div></center><br></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>&nbsp;&nbsp;&nbsp;In <ins class="diffchange diffchange-inline">further tests</ins>, we observed a significant <ins class="diffchange diffchange-inline">decrease </ins>in fluorescence when the cell type pLac+RFP+TetA was grown in tetracycline (50ug/mL) media. The cells continued to live in the tetracycline media with a marginal drop in cell density. However, the cells displaced considerably less fluorescence in this media even when induced with <ins class="diffchange diffchange-inline">IPTG</ins>. <ins class="diffchange diffchange-inline">This unexpected result suggests </ins>that <ins class="diffchange diffchange-inline">growth media affects </ins>cell fluorescence. </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>&nbsp;&nbsp;&nbsp;In <del class="diffchange diffchange-inline">addition</del>, we observed a significant <del class="diffchange diffchange-inline">reduction </del>in fluorescence when the cell type pLac+RFP+TetA was grown in tetracycline (50ug/mL) media. The cells continued to live in the tetracycline media with a marginal drop in cell density. However, the cells displaced considerably less fluorescence in this media even when induced with <del class="diffchange diffchange-inline">iPTG</del>. <del class="diffchange diffchange-inline"> These data imply </del>that <del class="diffchange diffchange-inline">the inducer did increase gene expression even if overall </del>cell <del class="diffchange diffchange-inline">density decreased. The construct fluoresced very little in comparison with the positive control (pLac+RBS+RFP) no matter what the conditions. The data support the hypothesis that there is a transcriptional terminator in the TetA gene. We performed another battery of tests to compare the pLac+TetA+RFP construct to a construct with the pLac+RFP+TetA orientation. When compared to cells with RFP followed by TetA, the construct’s negligible </del>fluorescence <del class="diffchange diffchange-inline">was even more apparent. Interestingly, the cells with the flipped orientation grew poorly in the presence of tetracycline but had a high fluorescence per cell. One possible explanation for this phenomenon is that the fluorescence is divided by such a low number of cells that small variations are magnified. Alternatively, the presence of tetracycline could be selecting for cells with high expression levels of both gene products. These few cells are producing high levels of TetA effluent pumps in order to survive. Correspondingly, they are producing a lot of RFP</del>.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><center><img src="https://static.igem.org/mediawiki/2010/6/61/Davidson-MissouriW_RFP_in_JM_109_vs._MG_1655_%282%29.png" id=fig3 height=600 width=800 ></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><center><img src="https://static.igem.org/mediawiki/2010/6/61/Davidson-MissouriW_RFP_in_JM_109_vs._MG_1655_%282%29.png" id=fig3 height=600 width=800 ></div></td></tr>
</table>Anrainahttp://2010.igem.org/wiki/index.php?title=Team:Davidson-MissouriW/MeasuringExpression&diff=204148&oldid=prevAnraina at 01:28, 28 October 20102010-10-28T01:28:18Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Results from past Davidson and Missouri Western research hinted at the presence of a transcriptional terminator within the TetA gene. Any gene located downstream of the 3’ end of the TetA gene did not appear to be expressed. This gene codes for an effluent pump that physically removes the antibody tetracycline from the cell. To test for the presence of this terminator, we created a construct with a fluorescent protein gene downstream of TetA. If TetA really does block transcription, we would expect to see no fluorescence despite induction of the promoter that precedes the construct (pLac+TetA+RFP).</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Results from past Davidson and Missouri Western research hinted at the presence of a transcriptional terminator within the TetA gene. Any gene located downstream of the 3’ end of the TetA gene did not appear to be expressed. This gene codes for an effluent pump that physically removes the antibody tetracycline from the cell. To test for the presence of this terminator, we created a construct with a fluorescent protein gene downstream of TetA. If TetA really does block transcription, we would expect to see no fluorescence despite induction of the promoter that precedes the construct (pLac+TetA+RFP).</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><center></p><img src="https://static.igem.org/mediawiki/2010/7/70/Normailized.png" ></center><br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><center></p><img src="https://static.igem.org/mediawiki/2010/7/70/Normailized.png" ></center><br></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;The data suggests that the order of genes (TetA and RFP) is important as pLac+RFP+TetA shows more fluorescence in comparison to pLac+TetA+RFP. The trend is reproducible when induced by IPTG (0.5mM) as well. <del class="diffchange diffchange-inline">In addition</del>, <del class="diffchange diffchange-inline">the construct with TetA before RFP was only able to survive the presence of tetracycline (50ug/mL) when the inducer IPTG was present. These data imply that the inducer did increase gene expression even if overall cell density decreased. The construct </del>fluoresced <del class="diffchange diffchange-inline">very little in comparison with </del>the positive control <del class="diffchange diffchange-inline">(</del>pLac<del class="diffchange diffchange-inline">+RBS</del>+RFP<del class="diffchange diffchange-inline">) no matter what the conditions</del>. The data <del class="diffchange diffchange-inline">support the </del>hypothesis that <del class="diffchange diffchange-inline">there is </del>a transcriptional terminator in the TetA gene.</p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;The data suggests that the order of genes (TetA and RFP) is important as pLac+RFP+TetA shows more fluorescence in comparison to pLac+TetA+RFP. The trend is reproducible when induced by IPTG (0.5mM) as well. <ins class="diffchange diffchange-inline">Both cell types, however</ins>, fluoresced <ins class="diffchange diffchange-inline">less than </ins>the positive control pLac+RFP. The data <ins class="diffchange diffchange-inline">also supports our </ins>hypothesis that a transcriptional terminator <ins class="diffchange diffchange-inline">is present </ins>in the TetA gene. </p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><center><img src="https://static.igem.org/mediawiki/2010/3/32/Davidson-MissouriWTPS3.png" id="fig2" height=600 width=800 ></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><center><img src="https://static.igem.org/mediawiki/2010/3/32/Davidson-MissouriWTPS3.png" id="fig2" height=600 width=800 ></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></center><br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></center><br></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>&nbsp;&nbsp;&nbsp;We performed another battery of tests to compare the pLac+TetA+RFP construct to a construct with the pLac+RFP+TetA orientation. When compared to cells with RFP followed by TetA, the construct’s negligible fluorescence was even more apparent. Interestingly, the cells with the flipped orientation grew poorly in the presence of tetracycline but had a high fluorescence per cell. One possible explanation for this phenomenon is that the fluorescence is divided by such a low number of cells that small variations are magnified. Alternatively, the presence of tetracycline could be selecting for cells with high expression levels of both gene products. These few cells are producing high levels of TetA effluent pumps in order to survive. Correspondingly, they are producing a lot of RFP.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>&nbsp;&nbsp;&nbsp;<ins class="diffchange diffchange-inline">In addition, we observed a significant reduction in fluorescence when the cell type pLac+RFP+TetA was grown in tetracycline (50ug/mL) media. The cells continued to live in the tetracycline media with a marginal drop in cell density. However, the cells displaced considerably less fluorescence in this media even when induced with iPTG. These data imply that the inducer did increase gene expression even if overall cell density decreased. The construct fluoresced very little in comparison with the positive control (pLac+RBS+RFP) no matter what the conditions. The data support the hypothesis that there is a transcriptional terminator in the TetA gene. </ins>We performed another battery of tests to compare the pLac+TetA+RFP construct to a construct with the pLac+RFP+TetA orientation. When compared to cells with RFP followed by TetA, the construct’s negligible fluorescence was even more apparent. Interestingly, the cells with the flipped orientation grew poorly in the presence of tetracycline but had a high fluorescence per cell. One possible explanation for this phenomenon is that the fluorescence is divided by such a low number of cells that small variations are magnified. Alternatively, the presence of tetracycline could be selecting for cells with high expression levels of both gene products. These few cells are producing high levels of TetA effluent pumps in order to survive. Correspondingly, they are producing a lot of RFP.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><center><img src="https://static.igem.org/mediawiki/2010/6/61/Davidson-MissouriW_RFP_in_JM_109_vs._MG_1655_%282%29.png" id=fig3 height=600 width=800 ></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><center><img src="https://static.igem.org/mediawiki/2010/6/61/Davidson-MissouriW_RFP_in_JM_109_vs._MG_1655_%282%29.png" id=fig3 height=600 width=800 ></div></td></tr>
</table>Anrainahttp://2010.igem.org/wiki/index.php?title=Team:Davidson-MissouriW/MeasuringExpression&diff=203326&oldid=prevAnraina at 01:03, 28 October 20102010-10-28T01:03:19Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;The system we originally designed to address the knapsack problem had several vital components. The TetA and RFP genes, along with their corresponding gene products, served as our reporters to see if and how a cell solved the knapsack problem. However, we recorded several observations regarding gene expression as we progressed. Characterizing and attempting to understand these foundational problems became one of the new focuses of this team.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;The system we originally designed to address the knapsack problem had several vital components. The TetA and RFP genes, along with their corresponding gene products, served as our reporters to see if and how a cell solved the knapsack problem. However, we recorded several observations regarding gene expression as we progressed. Characterizing and attempting to understand these foundational problems became one of the new focuses of this team.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Results from past Davidson and Missouri Western research hinted at the presence of a transcriptional terminator within the TetA gene. Any gene located downstream of the 3’ end of the TetA gene did not appear to be expressed. This gene codes for an effluent pump that physically removes the antibody tetracycline from the cell.<del class="diffchange diffchange-inline"></p></del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Results from past Davidson and Missouri Western research hinted at the presence of a transcriptional terminator within the TetA gene. Any gene located downstream of the 3’ end of the TetA gene did not appear to be expressed. This gene codes for an effluent pump that physically removes the antibody tetracycline from the cell. To test for the presence of this terminator, we created a construct with a fluorescent protein gene downstream of TetA. If TetA really does block transcription, we would expect to see no fluorescence despite induction of the promoter that precedes the construct (pLac+TetA+RFP).</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"><p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</del>To test for the presence of this terminator, we created a construct with a fluorescent protein gene downstream of TetA. If TetA really does block transcription, we would expect to see no fluorescence despite induction of the promoter that precedes the construct (pLac+TetA+RFP)<del class="diffchange diffchange-inline">.</del>.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><center></p><img src="https://static.igem.org/mediawiki/2010/7/70/Normailized.png" ></center><br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><center></p><img src="https://static.igem.org/mediawiki/2010/7/70/Normailized.png" ></center><br></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;The data suggests that the order of genes (TetA and RFP) is important as pLac+RFP+TetA shows more fluorescence in comparison to pLac+TetA+RFP. The trend is reproducible when induced by IPTG (0.5mM) as well. In addition, the construct with TetA before RFP was only able to survive the presence of tetracycline (50ug/mL) when the inducer IPTG was present. These data imply that the inducer did increase gene expression even if overall cell density decreased. The construct fluoresced very little in comparison with the positive control (pLac+RBS+RFP) no matter what the conditions. The data support the hypothesis that there is a transcriptional terminator in the TetA gene.</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;The data suggests that the order of genes (TetA and RFP) is important as pLac+RFP+TetA shows more fluorescence in comparison to pLac+TetA+RFP. The trend is reproducible when induced by IPTG (0.5mM) as well. In addition, the construct with TetA before RFP was only able to survive the presence of tetracycline (50ug/mL) when the inducer IPTG was present. These data imply that the inducer did increase gene expression even if overall cell density decreased. The construct fluoresced very little in comparison with the positive control (pLac+RBS+RFP) no matter what the conditions. The data support the hypothesis that there is a transcriptional terminator in the TetA gene.</p></div></td></tr>
</table>Anrainahttp://2010.igem.org/wiki/index.php?title=Team:Davidson-MissouriW/MeasuringExpression&diff=203271&oldid=prevAnraina at 01:01, 28 October 20102010-10-28T01:01:22Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;The system we originally designed to address the knapsack problem had several vital components. The TetA and RFP genes, along with their corresponding gene products, served as our reporters to see if and how a cell solved the knapsack problem. However, we recorded several observations regarding gene expression as we progressed. Characterizing and attempting to understand these foundational problems became one of the new focuses of this team.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;The system we originally designed to address the knapsack problem had several vital components. The TetA and RFP genes, along with their corresponding gene products, served as our reporters to see if and how a cell solved the knapsack problem. However, we recorded several observations regarding gene expression as we progressed. Characterizing and attempting to understand these foundational problems became one of the new focuses of this team.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Results from past Davidson and Missouri Western research hinted at the presence of a transcriptional terminator within the TetA gene. Any gene located downstream of the 3’ end of the TetA gene did not appear to be expressed. This gene codes for an effluent pump that physically removes the antibody tetracycline from the cell.</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Results from past Davidson and Missouri Western research hinted at the presence of a transcriptional terminator within the TetA gene. Any gene located downstream of the 3’ end of the TetA gene did not appear to be expressed. This gene codes for an effluent pump that physically removes the antibody tetracycline from the cell.</p></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;To test for the presence of this terminator, we created a construct with a fluorescent protein gene downstream of TetA. If TetA really does block transcription, we would expect to see no fluorescence despite induction of the promoter that precedes the construct (pLac+TetA+RFP)<del class="diffchange diffchange-inline">. We also tested pLac+TetA+RFP to see if the gene products provided protection from tetracycline compared to a negative control</del>..</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;To test for the presence of this terminator, we created a construct with a fluorescent protein gene downstream of TetA. If TetA really does block transcription, we would expect to see no fluorescence despite induction of the promoter that precedes the construct (pLac+TetA+RFP)..</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><center></p><img src="https://static.igem.org/mediawiki/2010/7/70/Normailized.png" ></center><br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><center></p><img src="https://static.igem.org/mediawiki/2010/7/70/Normailized.png" ></center><br></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"><center></p><img src="https://static.igem.org/mediawiki/2010/9/91/Wiki3.png" ></del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;The data <ins class="diffchange diffchange-inline">suggests </ins>that the <ins class="diffchange diffchange-inline">order of genes (TetA and RFP) is important as pLac+RFP+TetA shows more fluorescence in comparison to pLac+TetA+RFP. The trend is reproducible when induced by </ins>IPTG (0.5mM) <ins class="diffchange diffchange-inline">as well</ins>. In addition, the construct with TetA before RFP was only able to survive the presence of tetracycline (50ug/mL) when the inducer IPTG was present. These data imply that the inducer did increase gene expression even if overall cell density decreased. The construct fluoresced very little in comparison with the positive control (pLac+RBS+RFP) no matter what the conditions. The data support the hypothesis that there is a transcriptional terminator in the TetA gene.</p></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"></center><br></del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;The data <del class="diffchange diffchange-inline">suggest </del>that the IPTG (0.5mM) <del class="diffchange diffchange-inline">inducer decreases cell density in all three cell types</del>. In addition, the construct with TetA before RFP was only able to survive the presence of tetracycline (50ug/mL) when the inducer IPTG was present. These data imply that the inducer did increase gene expression even if overall cell density decreased. The construct fluoresced very little in comparison with the positive control (pLac+RBS+RFP) no matter what the conditions. The data support the hypothesis that there is a transcriptional terminator in the TetA gene.</p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><center><img src="https://static.igem.org/mediawiki/2010/3/32/Davidson-MissouriWTPS3.png" id="fig2" height=600 width=800 ></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><center><img src="https://static.igem.org/mediawiki/2010/3/32/Davidson-MissouriWTPS3.png" id="fig2" height=600 width=800 ></div></td></tr>
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</table>Anrainahttp://2010.igem.org/wiki/index.php?title=Team:Davidson-MissouriW/MeasuringExpression&diff=202985&oldid=prevAnraina at 00:49, 28 October 20102010-10-28T00:49:50Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Results from past Davidson and Missouri Western research hinted at the presence of a transcriptional terminator within the TetA gene. Any gene located downstream of the 3’ end of the TetA gene did not appear to be expressed. This gene codes for an effluent pump that physically removes the antibody tetracycline from the cell.</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Results from past Davidson and Missouri Western research hinted at the presence of a transcriptional terminator within the TetA gene. Any gene located downstream of the 3’ end of the TetA gene did not appear to be expressed. This gene codes for an effluent pump that physically removes the antibody tetracycline from the cell.</p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;To test for the presence of this terminator, we created a construct with a fluorescent protein gene downstream of TetA. If TetA really does block transcription, we would expect to see no fluorescence despite induction of the promoter that precedes the construct (pLac+TetA+RFP). We also tested pLac+TetA+RFP to see if the gene products provided protection from tetracycline compared to a negative control..</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;To test for the presence of this terminator, we created a construct with a fluorescent protein gene downstream of TetA. If TetA really does block transcription, we would expect to see no fluorescence despite induction of the promoter that precedes the construct (pLac+TetA+RFP). We also tested pLac+TetA+RFP to see if the gene products provided protection from tetracycline compared to a negative control..</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><center></p><img src="https://static.igem.org/mediawiki/2010/<del class="diffchange diffchange-inline">8</del>/<del class="diffchange diffchange-inline">8a</del>/<del class="diffchange diffchange-inline">Lbamp</del>.png" ></center><br></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><center></p><img src="https://static.igem.org/mediawiki/2010/<ins class="diffchange diffchange-inline">7</ins>/<ins class="diffchange diffchange-inline">70</ins>/<ins class="diffchange diffchange-inline">Normailized</ins>.png" ></center><br></div></td></tr>
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</table>Anrainahttp://2010.igem.org/wiki/index.php?title=Team:Davidson-MissouriW/MeasuringExpression&diff=202905&oldid=prevAnraina at 00:46, 28 October 20102010-10-28T00:46:42Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Results from past Davidson and Missouri Western research hinted at the presence of a transcriptional terminator within the TetA gene. Any gene located downstream of the 3’ end of the TetA gene did not appear to be expressed. This gene codes for an effluent pump that physically removes the antibody tetracycline from the cell.</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Results from past Davidson and Missouri Western research hinted at the presence of a transcriptional terminator within the TetA gene. Any gene located downstream of the 3’ end of the TetA gene did not appear to be expressed. This gene codes for an effluent pump that physically removes the antibody tetracycline from the cell.</p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;To test for the presence of this terminator, we created a construct with a fluorescent protein gene downstream of TetA. If TetA really does block transcription, we would expect to see no fluorescence despite induction of the promoter that precedes the construct (pLac+TetA+RFP). We also tested pLac+TetA+RFP to see if the gene products provided protection from tetracycline compared to a negative control..</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;To test for the presence of this terminator, we created a construct with a fluorescent protein gene downstream of TetA. If TetA really does block transcription, we would expect to see no fluorescence despite induction of the promoter that precedes the construct (pLac+TetA+RFP). We also tested pLac+TetA+RFP to see if the gene products provided protection from tetracycline compared to a negative control..</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><center></p><img src="https://static.igem.org/mediawiki/2010/<del class="diffchange diffchange-inline">b</del>/<del class="diffchange diffchange-inline">b2</del>/<del class="diffchange diffchange-inline">Finalwikiimage</del>.png" ></center><br></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><center></p><img src="https://static.igem.org/mediawiki/2010/<ins class="diffchange diffchange-inline">8</ins>/<ins class="diffchange diffchange-inline">8a</ins>/<ins class="diffchange diffchange-inline">Lbamp</ins>.png" ></center><br></div></td></tr>
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</table>Anrainahttp://2010.igem.org/wiki/index.php?title=Team:Davidson-MissouriW/MeasuringExpression&diff=202789&oldid=prevAnraina at 00:42, 28 October 20102010-10-28T00:42:35Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Results from past Davidson and Missouri Western research hinted at the presence of a transcriptional terminator within the TetA gene. Any gene located downstream of the 3’ end of the TetA gene did not appear to be expressed. This gene codes for an effluent pump that physically removes the antibody tetracycline from the cell.</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Results from past Davidson and Missouri Western research hinted at the presence of a transcriptional terminator within the TetA gene. Any gene located downstream of the 3’ end of the TetA gene did not appear to be expressed. This gene codes for an effluent pump that physically removes the antibody tetracycline from the cell.</p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;To test for the presence of this terminator, we created a construct with a fluorescent protein gene downstream of TetA. If TetA really does block transcription, we would expect to see no fluorescence despite induction of the promoter that precedes the construct (pLac+TetA+RFP). We also tested pLac+TetA+RFP to see if the gene products provided protection from tetracycline compared to a negative control..</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;To test for the presence of this terminator, we created a construct with a fluorescent protein gene downstream of TetA. If TetA really does block transcription, we would expect to see no fluorescence despite induction of the promoter that precedes the construct (pLac+TetA+RFP). We also tested pLac+TetA+RFP to see if the gene products provided protection from tetracycline compared to a negative control..</div></td></tr>
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</table>Anrainahttp://2010.igem.org/wiki/index.php?title=Team:Davidson-MissouriW/MeasuringExpression&diff=202660&oldid=prevAnraina at 00:37, 28 October 20102010-10-28T00:37:48Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Results from past Davidson and Missouri Western research hinted at the presence of a transcriptional terminator within the TetA gene. Any gene located downstream of the 3’ end of the TetA gene did not appear to be expressed. This gene codes for an effluent pump that physically removes the antibody tetracycline from the cell.</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Results from past Davidson and Missouri Western research hinted at the presence of a transcriptional terminator within the TetA gene. Any gene located downstream of the 3’ end of the TetA gene did not appear to be expressed. This gene codes for an effluent pump that physically removes the antibody tetracycline from the cell.</p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;To test for the presence of this terminator, we created a construct with a fluorescent protein gene downstream of TetA. If TetA really does block transcription, we would expect to see no fluorescence despite induction of the promoter that precedes the construct (pLac+TetA+RFP). We also tested pLac+TetA+RFP to see if the gene products provided protection from tetracycline compared to a negative control..</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;To test for the presence of this terminator, we created a construct with a fluorescent protein gene downstream of TetA. If TetA really does block transcription, we would expect to see no fluorescence despite induction of the promoter that precedes the construct (pLac+TetA+RFP). We also tested pLac+TetA+RFP to see if the gene products provided protection from tetracycline compared to a negative control..</div></td></tr>
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</table>Anrainahttp://2010.igem.org/wiki/index.php?title=Team:Davidson-MissouriW/MeasuringExpression&diff=202527&oldid=prevAnraina at 00:33, 28 October 20102010-10-28T00:33:18Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;The data suggest that the IPTG (0.5mM) inducer decreases cell density in all three cell types. In addition, the construct with TetA before RFP was only able to survive the presence of tetracycline (50ug/mL) when the inducer IPTG was present. These data imply that the inducer did increase gene expression even if overall cell density decreased. The construct fluoresced very little in comparison with the positive control (pLac+RBS+RFP) no matter what the conditions. The data support the hypothesis that there is a transcriptional terminator in the TetA gene.</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;The data suggest that the IPTG (0.5mM) inducer decreases cell density in all three cell types. In addition, the construct with TetA before RFP was only able to survive the presence of tetracycline (50ug/mL) when the inducer IPTG was present. These data imply that the inducer did increase gene expression even if overall cell density decreased. The construct fluoresced very little in comparison with the positive control (pLac+RBS+RFP) no matter what the conditions. The data support the hypothesis that there is a transcriptional terminator in the TetA gene.</p></div></td></tr>
</table>Anrainahttp://2010.igem.org/wiki/index.php?title=Team:Davidson-MissouriW/MeasuringExpression&diff=202296&oldid=prevAnraina at 00:23, 28 October 20102010-10-28T00:23:59Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;To test for the presence of this terminator, we created a construct with a fluorescent protein gene downstream of TetA. If TetA really does block transcription, we would expect to see no fluorescence despite induction of the promoter that precedes the construct (pLac+TetA+RFP). We also tested pLac+TetA+RFP to see if the gene products provided protection from tetracycline compared to a negative control..</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;To test for the presence of this terminator, we created a construct with a fluorescent protein gene downstream of TetA. If TetA really does block transcription, we would expect to see no fluorescence despite induction of the promoter that precedes the construct (pLac+TetA+RFP). We also tested pLac+TetA+RFP to see if the gene products provided protection from tetracycline compared to a negative control..</div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;The data suggest that the IPTG (0.5mM) inducer decreases cell density in all three cell types. In addition, the construct with TetA before RFP was only able to survive the presence of tetracycline (50ug/mL) when the inducer IPTG was present. These data imply that the inducer did increase gene expression even if overall cell density decreased. The construct fluoresced very little in comparison with the positive control (pLac+RBS+RFP) no matter what the conditions. The data support the hypothesis that there is a transcriptional terminator in the TetA gene.</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;The data suggest that the IPTG (0.5mM) inducer decreases cell density in all three cell types. In addition, the construct with TetA before RFP was only able to survive the presence of tetracycline (50ug/mL) when the inducer IPTG was present. These data imply that the inducer did increase gene expression even if overall cell density decreased. The construct fluoresced very little in comparison with the positive control (pLac+RBS+RFP) no matter what the conditions. The data support the hypothesis that there is a transcriptional terminator in the TetA gene.</p></div></td></tr>
</table>Anraina