Difference between revisions of "Team:Harvard BioDesign/Description"

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         <h2> Basic Parts</h2>
 
         <h2> Basic Parts</h2>
 
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We used the titratable Rhamnose promoter <partinfo>BBa_K902065</partinfo> in our <i>fimH</i> plasmids, and were able to improve its characterization. Preliminary results in our agglutination assays led us to hypothesize that the promoter was leaky. There is some question is to whether Rhamose is actually titratable. That is, the effect cross the population of the inducer appears to be aincrasing linearly with the under. But there was evidence in the literature, that t single cell elver, the cell was either completely induced or completely induced. So it appears to be tithable on a population level, but you'r actually seeing no expression over some subset and 100% of another. We wanted to see if that was the case for the iGEM repost, wWe found an existing part that contained the Rhamnose promoter expressing GFP, and we induced with varying concentrations of RHmnos, for very small to a lot of Rhamnose. We perfumed flow symmetry to measure th e fluorescence of individual cells, and shows the distribution of the fluorescence.  We see in our data that there is an intermediate concentration of RHamnoes for which part of the population shows the same fluoresce as the population with no Rhamnose added. Interesting for our own purposes because we cannot control the production of fimH like we thought we could.
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We used the rhamnose promoter <partinfo>BBa_K902065</partinfo> in our <i>fimH</i> plasmid, with the intention of controlling the amount of FimH production. Prior research indicates that there is some question as to whether rhamnose is actually titratable. That is, the effect on the population of the inducer appears to be increasing linearly with the amount of inducer . But evidence in the literature suggests that at the single cell level, each cell was either completely induced or completely uninduced. Thus while rhamnose appears to be titratable on the population level, it is possible that we were seeing no expression over some subset of cells and 100% expression over another. We wanted to see if this was the case for BBa_K902065. We found an existing iGEM part that contained BBa_K902065 and expressed GFP. Using this part, we induced with varying concentrations of rhamnose. We performed flow cytometry to measure the fluorescence of individual cells. Flow cytometry shows us the distribution of the fluorescence.  Our data shows that there is an intermediate concentration of inducer for which part of the population shows the same fluorescence  as the control population with no inducer added. This result confirms that the that the rhamnose promoter is bimodal; that is, each cell is turned either completely off or completely on. Thus result is interesting for our results because it shows that we could not control the level of <i> fimH</i> production of each cell, but only that of the population as a whole.
 
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Revision as of 01:09, 19 September 2015


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Basic Parts

We used the rhamnose promoter BBa_K902065 in our fimH plasmid, with the intention of controlling the amount of FimH production. Prior research indicates that there is some question as to whether rhamnose is actually titratable. That is, the effect on the population of the inducer appears to be increasing linearly with the amount of inducer . But evidence in the literature suggests that at the single cell level, each cell was either completely induced or completely uninduced. Thus while rhamnose appears to be titratable on the population level, it is possible that we were seeing no expression over some subset of cells and 100% expression over another. We wanted to see if this was the case for BBa_K902065. We found an existing iGEM part that contained BBa_K902065 and expressed GFP. Using this part, we induced with varying concentrations of rhamnose. We performed flow cytometry to measure the fluorescence of individual cells. Flow cytometry shows us the distribution of the fluorescence. Our data shows that there is an intermediate concentration of inducer for which part of the population shows the same fluorescence as the control population with no inducer added. This result confirms that the that the rhamnose promoter is bimodal; that is, each cell is turned either completely off or completely on. Thus result is interesting for our results because it shows that we could not control the level of fimH production of each cell, but only that of the population as a whole.

Composite Parts

Part Collection