Difference between revisions of "Team:William and Mary/Description"
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Original work done by Michael Elowitz investigated stochasticity in gene expression at the single cell level using a dual-reporter fluorescent system [1]. In this method two constructs are created that both drive expression of a fluorescent protein using the same promoter, RBS, and terminator. One of these reporter constructs uses YFP and the other CFP. This dual reporter system is what allows the investigator to differentiate between the extrinsic factors of noise, which should affect both constructs equally, and the intrinsic factors, which will result in a difference in the ratio of CFP: YFP fluorescence. These constructs are then integrated (notebook link here) into specific sites on the genome and the fluorescence output measured using fluorescent imaging. </p> <p> | Original work done by Michael Elowitz investigated stochasticity in gene expression at the single cell level using a dual-reporter fluorescent system [1]. In this method two constructs are created that both drive expression of a fluorescent protein using the same promoter, RBS, and terminator. One of these reporter constructs uses YFP and the other CFP. This dual reporter system is what allows the investigator to differentiate between the extrinsic factors of noise, which should affect both constructs equally, and the intrinsic factors, which will result in a difference in the ratio of CFP: YFP fluorescence. These constructs are then integrated (notebook link here) into specific sites on the genome and the fluorescence output measured using fluorescent imaging. </p> <p> | ||
− | <div><p | + | <div><p><IMG SRC="https://static.igem.org/mediawiki/2015/e/ed/WmRaserHoriz.jpeg"></p><p> Panel A shows an example of intrinsic and extrinsic noise; both the ratio of the fluorescence of the two reporters (intrinsic) and the absolute fluorescence of the two reporters (extrinsic) changes. Panel B shows an example of extrinsic noise only; the ratio of the fluorescence of the two reporters stays constant while the absolute fluorescence changes. Figure taken from:[2].</p></div></p> |
<div><p>Once individual cell measurements have been taken for both CFP and YFP expression using confocal microscopy, the intrinsic noise measurements can be calculated (link to John’s part about this) and further analyzed.</p></div> | <div><p>Once individual cell measurements have been taken for both CFP and YFP expression using confocal microscopy, the intrinsic noise measurements can be calculated (link to John’s part about this) and further analyzed.</p></div> |
Revision as of 11:46, 18 September 2015
Integrator Suites
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Antibiotic Operons
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dCas9s
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gRNAs
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XFPs Under Various Promoters
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G^2
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