Difference between revisions of "Part:BBa K2332031"

Latest revision as of 22:06, 22 October 2017

GFP-SpyCatcher

We designed our GFP-SpyCatcher 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'-SpyTag construct variants to analyse their co-localization. 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 662

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 <partinfo>BBa_K2332031 short</partinfo>
-This construct is the fusion of GFP with SpyCatcher. This construct allows to directly observe through fluorescence microscopy its localization, and most importantly its co-localization with SpyTag variants. Particularly for our project,  it allows us to quantify its binding onto cell surface displayed Intimin'-SpyTag.
+We designed our GFP-SpyCatcher 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'-SpyTag construct variants to analyse their co-localization. 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.
 <!-- Add more about the biology of this part here