Difference between revisions of "Part:BBa R0082:Experience"
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We tested this promoter for use in our landmine-detection system. Our plan was to use this promoter together with a computationally designed TNT receptor and the hybrid Trz signal transduction protein, which has the EnvZ kinase domain, hence phosphorylates OmpR, activating this promoter, in the presence of TNT. To check that this promoter was working correctly, we attached it to GFP and assayed its activity in ''E. coli'' TOP10 cells as described on the [https://parts.igem.org/Measurement Registry Measurement Page]. (We also made a ''lacZ'' reporter version, BBa_K216010, so that we could do Miller assays as a backup, but at the time of writing we have not obtained good quantitative results from this). To induce the promoter, we used procaine, which activates the EnvZ kinase. Our results indicated that the promoter is activated by increasing levels of procaine from 0 to 15 mM, but there is very high basal activity under the conditions used, even in the absence of procaine. We also made an ''envZ'' mutant host strain using the KEIO collection of ''E. coli'' mutants, but at the time of writing, we have not yet repeated the tests using this host. | We tested this promoter for use in our landmine-detection system. Our plan was to use this promoter together with a computationally designed TNT receptor and the hybrid Trz signal transduction protein, which has the EnvZ kinase domain, hence phosphorylates OmpR, activating this promoter, in the presence of TNT. To check that this promoter was working correctly, we attached it to GFP and assayed its activity in ''E. coli'' TOP10 cells as described on the [https://parts.igem.org/Measurement Registry Measurement Page]. (We also made a ''lacZ'' reporter version, BBa_K216010, so that we could do Miller assays as a backup, but at the time of writing we have not obtained good quantitative results from this). To induce the promoter, we used procaine, which activates the EnvZ kinase. Our results indicated that the promoter is activated by increasing levels of procaine from 0 to 15 mM, but there is very high basal activity under the conditions used, even in the absence of procaine. We also made an ''envZ'' mutant host strain using the KEIO collection of ''E. coli'' mutants, but at the time of writing, we have not yet repeated the tests using this host. | ||
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[http://2010.igem.org/Team:Tokyo_Tech/Project/wolf_coli/New_Series_of_PompC ...see more about P''ompC'' series] | [http://2010.igem.org/Team:Tokyo_Tech/Project/wolf_coli/New_Series_of_PompC ...see more about P''ompC'' series] | ||
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+ | '''Characterization of OmpC promoter by chemotaxis''' | ||
+ | <!-- DON'T DELETE --><partinfo>BBa_K1412010 StartReviews</partinfo> | ||
+ | {|width='80%' style='border:1px solid gray' | ||
+ | |- | ||
+ | |width='10%'| | ||
+ | <I>XMU-China iGEM 2014</I> | ||
+ | |width='60%' valign='top'| | ||
+ | [[File:BBa_K1412010-2.png|500px]] | ||
+ | |||
+ | '''The plot of Moving radius versus Sucrose concentration. The four curves were measured after 10h, 11h, 12h and 16.5h respectively. | ||
+ | ''' | ||
+ | |||
+ | <br> | ||
+ | ''E.coli'' make use of the EnvZ/OmpR system to mediate signal transduction in response to environmental osmolarity changes. EnvZ, a histidine kinase, undergoes trans-autophosphorylation, then the high energy phosphoryl group is subsequently transferred to OmpR, a response regulator. | ||
+ | In the system, OmpR-controlled promoter (PompC) is involved in. The expression strength of PompC is depending upon the medium osmolarity. When medium osmolarity is increasing, the EnvZ will phosphorylate more OmpR into phosphorylated OmpR (OmpR-P), and more OmpR-P will result in stronger expression strength of PompC. In our circuitry design, ''CheZ'' is upstream regulated by PompC. As the osmotic pressure is increasing, the motile ability of the engineered E.coli keeps growing, resulting in it's suicide. | ||
+ | |} | ||
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Revision as of 14:59, 10 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_R0082
User Reviews
UNIQ96938c87f07f094d-partinfo-00000000-QINU
No review score entered. Edinburgh iGEM 2009 |
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UNIQ96938c87f07f094d-partinfo-00000002-QINU
UNIQ96938c87f07f094d-partinfo-00000003-QINU
- Characterization of new series of OmpC propmoters
Tokyo Tech iGEM2010 |
In order to characterize PompC(C) BBa_395301, PompC(CB) BBa395302 and PompC(CS1) BBa_395303, each promoter was attached to GFP and its transcriptional activity was measured through the GFP expression. [http://2010.igem.org/Team:Tokyo_Tech/Project/wolf_coli/New_Series_of_PompC ...see more about PompC series] |
UNIQ96938c87f07f094d-partinfo-00000004-QINU
Characterization of OmpC promoter by chemotaxis
UNIQ96938c87f07f094d-partinfo-00000005-QINU
XMU-China iGEM 2014 |
The plot of Moving radius versus Sucrose concentration. The four curves were measured after 10h, 11h, 12h and 16.5h respectively.
|
UNIQ96938c87f07f094d-partinfo-00000006-QINU