Difference between revisions of "Part:BBa K1460008:Experience"
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===Applications of BBa_K1460008=== | ===Applications of BBa_K1460008=== | ||
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| + | <html> | ||
| + | <script type="text/javascript"> | ||
| + | $(window).load(function() { | ||
| + | $('li.p_wet_metal').addClass('active'); | ||
| + | }); | ||
| + | </script> | ||
| + | <div class="container" id="top"> | ||
| + | <div class="row"> | ||
| + | <div class="col-md-12 col-xs-18"> | ||
| + | <h2>Cornell 2014 Results</h2> | ||
| + | <br> | ||
| + | Part <a href="https://parts.igem.org/Part:BBa_K1460005">BBa_K1460005</a> in pUC57 was co-transformed with part <a href="https://parts.igem.org/Part:BBa_K1460001">BBa_K1460001</a> (GST-YMT) in pSB1C3 and selected for with both ampicillin and chloramphenicol to effectively create the lead sequestration part BBa_K1460008. To test for sequestration efficiency, both BL21 and BL21 engineered with BBa_K1460001 and BBa_K1460005 were grown with LB + 0.1% Arabinose for 8 hours and then diluted by 1/2 with LB + 2 mM Pb for a final lead concentration of 1 mM. These cultures were grown for 8 more hours. The cells were then removed and supernatant was tested for lead concentration using Inductively Coupled Plasma Atomic Emission Spectroscopy (ICP-AES) with the help of Cornell's Nutrient Analysis Lab. Error bars in chart represent standard deviation of three biological replicates. | ||
| + | </div> | ||
| + | </div> | ||
| + | <div class="row"> | ||
| + | <div class="col-md-6 col-xs-9"> | ||
| + | <img src="https://static.igem.org/mediawiki/2014/b/bf/Cornell_final_lead_concentration.png" width="100%"> | ||
| + | </div> | ||
| + | <div class="row"> | ||
| + | <div class="col-md-6 col-xs-9"> | ||
| + | <img src="https://static.igem.org/mediawiki/2014/5/57/Cornell_Lead_per_OD.png" width="100%"> | ||
| + | </div> | ||
| + | </div> | ||
| + | <div class="row"> | ||
| + | <div class="col-md-12 col-xs-18"> | ||
| + | The chart on the top shows the average final concentration of lead in the cultures. There was no statistically significant difference between BL21 and BL21 engineered with BBa_K1460001 and BBa_K1460005. However, when we consider cell density and plot the amount of metal removed per OD (on the bottom chart) there is a statistically significant difference between the two strains. This data suggests that cells engineered with CBP4 and GST-YMT are in fact able to remove lead ions from water. | ||
| + | <br><br> | ||
| + | Ideally, this experiment would be run with the OD of both strains remaining the same to prevent changes in metabolite concentrations. This is difficult in this experiment as, <a href="https://parts.igem.org/Part:BBa_K1460001:Experience">as we have shown</a>, cells expressing metallothionein have inhibited growth. | ||
| + | </div> | ||
| + | </div> | ||
| + | </body> | ||
| + | </html> | ||
===User Reviews=== | ===User Reviews=== | ||
Revision as of 17:43, 14 October 2014
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_K1460008
Cornell 2014 Results
Part BBa_K1460005 in pUC57 was co-transformed with part BBa_K1460001 (GST-YMT) in pSB1C3 and selected for with both ampicillin and chloramphenicol to effectively create the lead sequestration part BBa_K1460008. To test for sequestration efficiency, both BL21 and BL21 engineered with BBa_K1460001 and BBa_K1460005 were grown with LB + 0.1% Arabinose for 8 hours and then diluted by 1/2 with LB + 2 mM Pb for a final lead concentration of 1 mM. These cultures were grown for 8 more hours. The cells were then removed and supernatant was tested for lead concentration using Inductively Coupled Plasma Atomic Emission Spectroscopy (ICP-AES) with the help of Cornell's Nutrient Analysis Lab. Error bars in chart represent standard deviation of three biological replicates.
The chart on the top shows the average final concentration of lead in the cultures. There was no statistically significant difference between BL21 and BL21 engineered with BBa_K1460001 and BBa_K1460005. However, when we consider cell density and plot the amount of metal removed per OD (on the bottom chart) there is a statistically significant difference between the two strains. This data suggests that cells engineered with CBP4 and GST-YMT are in fact able to remove lead ions from water.
Ideally, this experiment would be run with the OD of both strains remaining the same to prevent changes in metabolite concentrations. This is difficult in this experiment as, as we have shown, cells expressing metallothionein have inhibited growth.
Ideally, this experiment would be run with the OD of both strains remaining the same to prevent changes in metabolite concentrations. This is difficult in this experiment as, as we have shown, cells expressing metallothionein have inhibited growth.