Team:UFMG Brazil/Results
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- | The results shown in here were performed as described in <html><a href='https://2013.igem.org/Team:UFMG_Brazil/Protocols'>Protocols</a></html>. We used Varioskan Flash Multimode Reader (Thermo Scientific™) to do the reads of YFP constructs and Synergy 2 (Biotek) to do the reads of | + | The results shown in here were performed as described in <html><a href='https://2013.igem.org/Team:UFMG_Brazil/Protocols'>Protocols</a></html>. We used Varioskan Flash Multimode Reader (Thermo Scientific™) to do the reads of YFP constructs and Synergy 2 (Biotek) to do the reads of RFP constructs. |
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'''PSB1C3_TorCAD+RFP:''' | '''PSB1C3_TorCAD+RFP:''' | ||
- | We have constructed the composite TorCAD + RFP to detect TMAO (Trimethylamine N-oxide) in the serum of patients with cardiac risk. These patients present more TMAO than normal individuals. | + | We have constructed the composite TorCAD + RFP <html><a href=http://parts.igem.org/cgi/partsdb/pgroup.cgi?pgroup=iGEM2013&group=UFMG_Brazil>Ba_K1086002</a></html> to detect TMAO (Trimethylamine N-oxide) in the serum of patients with cardiac risk. These patients present more TMAO than normal individuals. |
- | To prove that our construct works, we made some tests with transformed ''E. coli'' XL1-Blue. We added different concentrations of TMAO to bacterial cultures and measured their fluorescence and absorbance for | + | To prove that our construct works, we made some tests with transformed ''E. coli'' XL1-Blue. We added different concentrations of TMAO to bacterial cultures and measured their fluorescence and absorbance for 15 hours. The fluorescence stars to increase only 8 hours after the treatment with TMAO (Figures 10 and 11) and continue to increase until 13 hours after the treatment. We could observe fluorescence appearence only with the concentration of 100 μM of TMAO. |
- | [[File:clone2_tmao_fluo.jpg|600px|thumb|center|'''Figure 10: | + | [[File:clone2_tmao_fluo.jpg|600px|thumb|center|'''Figure 10: Fluorometric reads of cultures ''of E. coli'' XL1-Blue carrying the plasmid PSB1C3_TorCAD + RFP, after treatment with different concentrations of TMAO.''' Bacteria were treated with 0 μM, 1 μM, 10 μM, 100 μM, 1 mM, 10 mM and 100 mM. After that, fluorescence was read hourly, until 15 hours. The beginning of the fluorescence increase can be seen about 8 hours after the treatment.]] |
- | [[File:clone2_tmao_fluo_per_abs.jpg|600px|thumb|center|'''Figure 11: | + | [[File:clone2_tmao_fluo_per_abs.jpg|600px|thumb|center|'''Figure 11: Fluorometric and absorbance reads of cultures of ''E. coli'' XL1-Blue carrying the plasmid PSB1C3_TorCAD + RFP, after treatment with different concentrations of TMAO.''' The fluorescence reads shown in figure 11 divided by the absorbance, resulting in the graphic above.]] |
- | == | + | == Conclusions and Perspectives == |
+ | During the time of the competition our team was able to: | ||
- | + | * Modify the previous described biobrick <html><a href=http://parts.igem.org/Part:BBa_K540001>BBa_K540001</a></html> adding a fluorescent yellow protein and give it a new functionality. Our biobrick <html><a href=http://parts.igem.org/cgi/partsdb/pgroup.cgi?pgroup=iGEM2013&group=UFMG_Brazil>BBa_K1086001</a></html> is able to differentiate serum from isquemic and non isquemic mice. | |
- | + | * Generate a new Biobrick <html><a href=http://parts.igem.org/cgi/partsdb/pgroup.cgi?pgroup=iGEM2013&group=UFMG_Brazil>BBa_K1086002</a></html> able to detect a concentration of 100 µM of TMAO. | |
- | + | * Generate modeling that describe our systems and simulate it. | |
- | + | * Expand the knowledge about synthetic biology in our University, our State and in Brazil. | |
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+ | As a perspective of this work, we aim: | ||
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+ | * To incorporate more plasmid copies with Tor_CAD <html><a href=http://parts.igem.org/cgi/partsdb/pgroup.cgi?pgroup=iGEM2013&group=UFMG_Brazil>BBa_K1086002</a></html> biobrick to decrease the time to detect TMAO and to detect less TMAO concentrations. | ||
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+ | * To test our 2 constructs with isquemic and non isquemic pacients serum. | ||
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+ | * To test our 2 constructs with serum from obese and non-obese patients to try to predict the potential risk of cardiovascular events in these patients. | ||
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{{Team:UFMG Brazil/sponsor}} | {{Team:UFMG Brazil/sponsor}} |
Latest revision as of 01:08, 29 October 2013