Difference between revisions of "Part:BBa K2332017"

Latest revision as of 21:51, 22 October 2017

GFP-SpyTag

We designed our GFP-SpyTag Biobrick to test bacterial cell adhesion through fluorescence microscopy. This part produces the GFP-SpyTag fusion protein, 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.

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 644

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 <partinfo>BBa_K2332017 short</partinfo>
-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).
+We designed our GFP-SpyTag Biobrick to test bacterial cell adhesion through fluorescence microscopy. This part produces the GFP-SpyTag fusion protein, 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.
-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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