Difference between revisions of "Part:BBa K2582000"
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<partinfo>BBa_K2582000 short</partinfo>
 
<partinfo>BBa_K2582000 short</partinfo>
  
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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. However, it alone does not fluoresce well in E. coli, possibly due to low folding efficiency.
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This Biobrick contains the DNA sequence of the light-up RNA Aptamer, 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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miniSpinach can be in vitro transcribed or expressed in E. coli.
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Mixing the Spinach aptamer with excess DFHBI yielded a 100-fold increase in the fluorescence level of the reaction mixture. However, its fluorescence in E. coli is hard to visualize and quite close to the E. coli autofluorescence level. Upon comparison between In-cell, total RNA and in vitro synthesized RNA, it is found that miniSpinach does not fold well in E. coli.
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K2582000 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K2582000 SequenceAndFeatures</partinfo>

Revision as of 10:07, 14 October 2018


miniSpinach (P1-a5-b3)

This Biobrick contains the DNA sequence 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

miniSpinach can be in vitro transcribed or expressed in E. coli.

Mixing the Spinach aptamer with excess DFHBI yielded a 100-fold increase in the fluorescence level of the reaction mixture. However, its fluorescence in E. coli is hard to visualize and quite close to the E. coli autofluorescence level. Upon comparison between In-cell, total RNA and in vitro synthesized RNA, it is found that miniSpinach does not fold well in E. coli.


Sequence and Features


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