Difference between revisions of "Part:BBa K2582011"

 
 
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<partinfo>BBa_K2582011 short</partinfo>
 
<partinfo>BBa_K2582011 short</partinfo>
  
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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].
  
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===Usage and Biology===
 
===Usage and Biology===
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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.
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===Characterization===
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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.]]
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K2582011 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K2582011 SequenceAndFeatures</partinfo>

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.

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.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]