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Team:ETHZ Basel/Biology/Status
From 2010.igem.org
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= testing of biobricks = | = testing of biobricks = | ||
| - | == testing of che-fusions == | + | === testing of che-fusions === |
We're currently trying to establish an assay to test for the chemotactic behavour of ''E. coli''. A first try with a "syringe test" failed. We'll redo the test and are looking for other possibilites. Maybe a swarm test? | We're currently trying to establish an assay to test for the chemotactic behavour of ''E. coli''. A first try with a "syringe test" failed. We'll redo the test and are looking for other possibilites. Maybe a swarm test? | ||
*tested biobricks: che-fusions | *tested biobricks: che-fusions | ||
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*positive control: wild type ''E. coli'' | *positive control: wild type ''E. coli'' | ||
| - | == testing of localicer-fusions == | + | === testing of localicer-fusions === |
We are planning to use a fluorescence microscope in order to visualize the distribution of localizer-cyFP fusions within a single ''E. coli''. | We are planning to use a fluorescence microscope in order to visualize the distribution of localizer-cyFP fusions within a single ''E. coli''. | ||
| - | == testing of couplers == | + | === testing of couplers === |
We're planning to use the setup as for the testing of the localizers. | We're planning to use the setup as for the testing of the localizers. | ||
*tested biobricks: localizer-PhyB and PIF3-cyFP, localizer-PIF3 and PhyB-cyFP | *tested biobricks: localizer-PhyB and PIF3-cyFP, localizer-PIF3 and PhyB-cyFP | ||
Revision as of 15:12, 6 August 2010
generation of subparts
| Protein | primers | PCR product | library of transformants | single colonies | storage vector | insert | |
|---|---|---|---|---|---|---|---|
| che proteins | CheY-A | recieved | purified on stock | on agar plate | not available | purified on stock | on stock |
| CheY-C | recieved | purified in stock | on agar plate | not available | purified on stock | on stock | |
| CheB-A | recieved | purified in stock | on agar plate | 3 on agar plate | purified on stock | on stock | |
| CheB-C | recieved | purified in stock | on agar plate | 3 on agar plate | purified on stock | on stock | |
| CheR-A | recieved | purified in stock | on agar plate | 3 on agar plate | digest has to be redone | ||
| CheR-C | recieved | purified in stock | on agar plate | 3 on agar plate | digest has to be redone | ||
| localizers | TrigF-A | recieved | purified in stock | on agar plate | 3 on agar plate | purified on stock | |
| MreB-C | recieved | purified in stock | in process | ||||
| tetR-A | recieved | purified in stock | on agar plate | 3 on agar plate | purified on stock | on stock | |
| couplers | PIF3-A | recieved | purified in stock | on agar plate | 3 on agar plate | purified on stock | |
| PIF3-C | recieved | purified in stock | on agar plate | 3 on agar plate | purified on stock | ||
| PhyB-C | recieved | purified in stock | on agar plate | 3 on agar plate | purified on stock | ||
| reporters | cyFP-A | recieved | purified in stock | on agar plate | 3 on agar plate | purified on stock | on stock |
| cyFP-C | recieved | purified in stock | on agar plate | 3 on agar plate | purified on stock |
generation of biobricks
We'll talk to Fabian. Apparently he has a nice protocol to generate acceptor vectors and transform E. coli sequentially in a 96 well plate.
testing of biobricks
testing of che-fusions
We're currently trying to establish an assay to test for the chemotactic behavour of E. coli. A first try with a "syringe test" failed. We'll redo the test and are looking for other possibilites. Maybe a swarm test?
- tested biobricks: che-fusions
- negative control: E. coli mutated for the accoding che-protein
- positive control: wild type E. coli
testing of localicer-fusions
We are planning to use a fluorescence microscope in order to visualize the distribution of localizer-cyFP fusions within a single E. coli.
testing of couplers
We're planning to use the setup as for the testing of the localizers.
- tested biobricks: localizer-PhyB and PIF3-cyFP, localizer-PIF3 and PhyB-cyFP
Images should look as in testing of localizer-fusions. Preliminary clarificaiton of the suitability of the approach are to be done.
bringing E. lemming alive
timetable
