Difference between revisions of "Part:BBa K2582011"
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<partinfo>BBa_K2582011 short</partinfo> | <partinfo>BBa_K2582011 short</partinfo> | ||
| − | This Biobrick contains the T7 RNA Polymerase-based expression cassette of the light-up RNA Aptamer, miniSpinach. | + | This Biobrick contains the T7 RNA Polymerase-based expression cassette of the light-up RNA Aptamer, [https://parts.igem.org/wiki/index.php?title=Part:BBa_K2582000 miniSpinach]. |
===Usage and Biology=== | ===Usage and Biology=== | ||
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===Characterization=== | ===Characterization=== | ||
The fluorescence of miniSpinach without scaffold in E. coli is hard to visualize and quite close to the E. coli autofluorescence level. Upon comparison between in-cell fluorescence and total RNA, it is found that miniSpinach does not fold well in E. coli. | The fluorescence of miniSpinach without scaffold in E. coli is hard to visualize and quite close to the E. coli autofluorescence level. Upon comparison between in-cell fluorescence and total RNA, it is found that miniSpinach does not fold well in E. coli. | ||
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[[File:BBa K2582000-Fig2.png|500px|thumb|left|Comparison of miniSpinach fluorescence between in-cell and extracted conditions. For total RNA fluorescence assay, the same RNA amount corresponding to OD600 for in-cell assay was used.]] | [[File:BBa K2582000-Fig2.png|500px|thumb|left|Comparison of miniSpinach fluorescence between in-cell and extracted conditions. For total RNA fluorescence assay, the same RNA amount corresponding to OD600 for in-cell assay was used.]] | ||
Latest revision as of 13:42, 16 October 2018
T7 Promoter-miniSpinach(P1-a5-b3)-T7 Terminator
This Biobrick contains the T7 RNA Polymerase-based expression cassette of the light-up RNA Aptamer, miniSpinach.
Usage and Biology
miniSpinach is a minimal version of the Spinach aptamer developed by Paige et. al. (2011). It is created previously by generating mutants of the Spinach aptamer by decreasing stem lengths (Ong et. al. 2017). When transcribed in vitro, it allows the docking of fluorogen 3,5-difluoro-4-hydroxybenzylidene imidazolinone (DFHBI) and increases its quantum yields by hundreds of folds.
Characterization
The fluorescence of miniSpinach without scaffold in E. coli is hard to visualize and quite close to the E. coli autofluorescence level. Upon comparison between in-cell fluorescence and total RNA, it is found that miniSpinach does not fold well in E. coli.
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]