Difference between revisions of "Part:BBa K1493501"
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This BioBrick is used by [http://2014.igem.org/Team:Wageningen_UR BananaGuard] as a control in the [http://2014.igem.org/Team:Wageningen_UR/project/characterization rhamnose mediated characterization]. | This BioBrick is used by [http://2014.igem.org/Team:Wageningen_UR BananaGuard] as a control in the [http://2014.igem.org/Team:Wageningen_UR/project/characterization rhamnose mediated characterization]. | ||
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| + | ===Usage and Biology=== | ||
https://static.igem.org/mediawiki/2014/f/f9/Wageningen_UR_killswitch_rhamnose.png <br> | https://static.igem.org/mediawiki/2014/f/f9/Wageningen_UR_killswitch_rhamnose.png <br> | ||
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''Figure 1. Average GFP fluorescence of E. coli DH5α carrying a pSB3K3 plasmid containing the pRha gfp BioBrick. At t=0 the cells were induced with different concentrations of rhamnose in triplo. Fluorescence is measured in a plate reader in time (A) and at time point 8.25 (B) normalized for OD600.'' | ''Figure 1. Average GFP fluorescence of E. coli DH5α carrying a pSB3K3 plasmid containing the pRha gfp BioBrick. At t=0 the cells were induced with different concentrations of rhamnose in triplo. Fluorescence is measured in a plate reader in time (A) and at time point 8.25 (B) normalized for OD600.'' | ||
| − | + | The measurements in Figure 1 indicate a tuneable activation of pRha by rhamnose. The RFU values of 0% and 0.001% rhamnose are not significant taking into account the high standard deviation for these measurements as can be seen in Figure 1B. From a rhamnose concentration of 0.01% to 0.2% a significant increase in fluorescence is measured. Fluorescence from Figure 2 can be used to predict the concentration of repressor protein produced. Data points for time 8.25 h were chosen for the graph in Figure 2B due to the peak at time for 0.2% rhamnose, which is visible in Figure 1. This peak can be explained by the rhamnose depletion, as it is consumed by E. coli, causing the stop of promoter induction. | |
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Revision as of 08:41, 23 October 2014
pRha + GFP
GFP controlled by pRha.
This BioBrick is used by [http://2014.igem.org/Team:Wageningen_UR BananaGuard] as a control in the [http://2014.igem.org/Team:Wageningen_UR/project/characterization rhamnose mediated characterization].
Usage and Biology
Figure 1. Average GFP fluorescence of E. coli DH5α carrying a pSB3K3 plasmid containing the pRha gfp BioBrick. At t=0 the cells were induced with different concentrations of rhamnose in triplo. Fluorescence is measured in a plate reader in time (A) and at time point 8.25 (B) normalized for OD600.
The measurements in Figure 1 indicate a tuneable activation of pRha by rhamnose. The RFU values of 0% and 0.001% rhamnose are not significant taking into account the high standard deviation for these measurements as can be seen in Figure 1B. From a rhamnose concentration of 0.01% to 0.2% a significant increase in fluorescence is measured. Fluorescence from Figure 2 can be used to predict the concentration of repressor protein produced. Data points for time 8.25 h were chosen for the graph in Figure 2B due to the peak at time for 0.2% rhamnose, which is visible in Figure 1. This peak can be explained by the rhamnose depletion, as it is consumed by E. coli, causing the stop of promoter induction.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 792