Team:Colombia Uniandes/Parts
From 2013.igem.org
(Difference between revisions)
(→Glucocorticoid sensor) |
Sylvita1015 (Talk | contribs) (→Glucocorticoid sensor) |
||
Line 66: | Line 66: | ||
==='''Characterization of reporters'''=== | ==='''Characterization of reporters'''=== | ||
- | To assess the strength of our promoters when induced with Dexamethasone, we performed a fluorometric assay using mCherry as our reporter.Since we don't have our transactivating protein ready yet, we co-transformed our parts with the | + | To assess the strength of our promoters when induced with Dexamethasone, we performed a fluorometric assay using mCherry as our reporter.Since we don't have our transactivating protein ready yet, we co-transformed our parts with the pAT7002 vector (Aoyama and Chua, 997, Plant J. 11 , p605-612), which contains a well characterized Glucocorticoid responsive element, similar to our proposed inductor, into ''E. coli'' DH10B cells. |
[[File:Response1hr.jpg|700px|thumb|center|''' Unsaturated curve where we see how the mCherry reporter appears after the addition of 10 uM dexamethasone, a glucocorticoid. Emmision intensity was measured at 607nm after excitation at 586nm''']] | [[File:Response1hr.jpg|700px|thumb|center|''' Unsaturated curve where we see how the mCherry reporter appears after the addition of 10 uM dexamethasone, a glucocorticoid. Emmision intensity was measured at 607nm after excitation at 586nm''']] |
Revision as of 03:23, 28 September 2013
Parts
Glucocorticoid sensor
Here are the parts sent to the registry:
Part Name | Registry Number |
E1 | BBa_K1144001 |
E2 | BBa_K1144002 |
E3 | BBa_K1144003 |
E4 | BBa_K1144004 |
E1GF | BBa_K1144005 |
E2GF | BBa_K1144006 |
E3GF | BBa_K1144007 |
E4GF | BBa_K1144008 |
You can see the confirmation gels of the parts below:
Characterization of reporters
To assess the strength of our promoters when induced with Dexamethasone, we performed a fluorometric assay using mCherry as our reporter.Since we don't have our transactivating protein ready yet, we co-transformed our parts with the pAT7002 vector (Aoyama and Chua, 997, Plant J. 11 , p605-612), which contains a well characterized Glucocorticoid responsive element, similar to our proposed inductor, into E. coli DH10B cells.
We also decided to visually inspect our induced transformants. Here two of the images taken using a epifluorescent microscopy with a TRITC filter.