Difference between revisions of "Part:BBa K2582017"
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===Characterization=== | ===Characterization=== | ||
| − | This aptamer can be expressed in <i>E. coli</i> DH5a under a constitutive lpp promoter. Comparing the 3 RNA reporters we constructed and the pre-existed Spinach2.1, | + | This aptamer can be expressed in <i>E. coli</i> DH5a under a constitutive lpp promoter. Comparing the 3 RNA reporters we constructed and the pre-existed Spinach2.1, tdBroccoli is moderately brighter. |
[[File:Improve a previous bobrick.jpg|400px|thumb|left|Fluorescence data obtained from plate reader. DH5a transformed with pSB1C3-lpp-Spinach2.1 or other Spinach variants were incubated overnight at 37 degrees Celsius. Overnight culture was normalized to OD=0.2 and incubated in 200uM DFHBI for 45 minutes. N=1. More replicates are needed.]] | [[File:Improve a previous bobrick.jpg|400px|thumb|left|Fluorescence data obtained from plate reader. DH5a transformed with pSB1C3-lpp-Spinach2.1 or other Spinach variants were incubated overnight at 37 degrees Celsius. Overnight culture was normalized to OD=0.2 and incubated in 200uM DFHBI for 45 minutes. N=1. More replicates are needed.]] | ||
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Latest revision as of 19:41, 16 October 2018
lpp promoter-tdBroccoli
This part is a constitutive expression cassette of a RNA reporter that can be used to measure promoter activity directly. It contains the light-up RNA Aptamer [http://doi.org/10.1021/ja508478x dimeric Broccoli]in the human tRNALys3 scaffold. This biobrick is an improvement from K1330000, which is brighter than the original design.
Usage and Biology
The Spinach aptamer and its derivatives allow the docking of fluorogen 3,5-difluoro-4-hydroxybenzylidene imidazolinone (DFHBI) and increases its quantum yields by hundreds of folds. By surrounding it with a tRNA scaffold, its folding in cell can be enhanced and its expression level can be stabilized.
Characterization
This aptamer can be expressed in E. coli DH5a under a constitutive lpp promoter. Comparing the 3 RNA reporters we constructed and the pre-existed Spinach2.1, tdBroccoli is moderately brighter.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]