Difference between revisions of "Part:BBa K2332053"

Revision as of 23:44, 22 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.

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

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 <partinfo>BBa_K2332053 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 variants.
+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.
-To use the GFP-Spytag, one needs to lyse recombinant E. coli cells and purify the protein extract in order to visualize its localization on the cell surface of cells expressing intimin'-SpyCatcher variants.
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