Difference between revisions of "Part:BBa K2332041"

Revision as of 14:26, 25 October 2017

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

This construct encodes the fusion of GFP with SpyCatcher under the control of Pblind, a blue light inducible promoter. Cells must also express EL222 (BBa_K2332004) for blue light induction.

To use the GFP-SpyCatcher, one needs to transform E. Coli cells with our Biobrick and expose to blue-light to allow the cells to produce the fusion protein. Then, cells are lysed and the protein purified. 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.

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 741

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K2332041 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.
+This construct encodes the fusion of GFP with SpyCatcher under the control of [https://parts.igem.org/Part:BBa_K2332002 Pblind], a blue light inducible promoter. Cells must also express EL222 ([https://parts.igem.org/Part:BBa_K2332004 BBa_K2332004]) for blue light induction.
+To use the GFP-SpyCatcher, one needs to transform E. Coli cells with our Biobrick and expose to blue-light to allow the cells to produce the fusion protein. Then, cells are lysed and the protein purified. 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.