Difference between revisions of "Part:BBa K2332000"

 
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We designed our GFP-SpyTag Biobrick to test bacterial cell adhesion through fluorescence microscopy. This part produces the GFP-SpyTag fusion protein, which we experimentally used in combination with our Intimin-SpyCatcher Biobrick (see experiment).
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We designed our GFP-SpyTag Biobrick to test bacterial cell adhesion through fluorescence microscopy. This part produces the GFP-SpyTag fusion protein constitutively, which can then be cell extracted and purified to use in combination with cells expressing our Intimin'-SpyCatcher construct variants. SpyTag (13 amino acids) and SpyCatcher (138 amino acids, 15 kDa) are protein binding partners that originate from CnaB2 (immunoglobulin-llike collagen adhesin domain) of the FbaB protein, found in the invasive strains of S. pyogenes.
  
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To use the GFP-Spytag, one needs to transform E. Coli cells with our Biobrick and allow the cells to produce the fusion protein. Then, cells are lysed and the protein purified to be used in the experiment.
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[[File:GFP-SpyTag.png|thumb|center|500px| Figure 1: E. coli cells constitutively expressing GFP-SpyTag]]
  
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
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===Functional Parameters===
 
===Functional Parameters===
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<partinfo>BBa_K2332000 parameters</partinfo>
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<partinfo>BBa_K2332053 parameters</partinfo>
 
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Revision as of 23:59, 23 October 2017


Blue light inducible expression of GFP-SpyTag (Pblind GFP-SpyTag)

We designed our GFP-SpyTag Biobrick to test bacterial cell adhesion through fluorescence microscopy. This part produces the GFP-SpyTag fusion protein constitutively, which can then be cell extracted and purified to use in combination with cells expressing our Intimin'-SpyCatcher construct variants. SpyTag (13 amino acids) and SpyCatcher (138 amino acids, 15 kDa) are protein binding partners that originate from CnaB2 (immunoglobulin-llike collagen adhesin domain) of the FbaB protein, found in the invasive strains of S. pyogenes.

Figure 1: E. coli cells constitutively expressing GFP-SpyTag

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
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 723