Difference between revisions of "Part:BBa K2332040"

Revision as of 13:59, 25 October 2017

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

This construct displays SpyTag fused to GFP on the cell surface of E. coli cells under the control of Pblind, a blue light inducible promoter. Cells must also express EL222 (BBa_K2332004). Upon blue-light (465nm) exposure, EL222 dimerises and binds to its DNA-binding region within Pblind to recruit RNAP and induce transcription of Intimin'-GFP-SpyTag.

Intimin' is a truncated version of Intimin (EaeA intimin), an outer membrane surface protein, which has shown to display proteins of similar size to GFP fused to its N terminal. This construct allows to directly observe through fluorescence microscopy the functionality of Intimin for cell surface display under blue light induction.

Sequence and Features

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal SpeI site found at 2844
12
INCOMPATIBLE WITH RFC[12]
Illegal SpeI site found at 2844
21
COMPATIBLE WITH RFC[21]
23
INCOMPATIBLE WITH RFC[23]
Illegal SpeI site found at 2844
25
INCOMPATIBLE WITH RFC[25]
Illegal SpeI site found at 2844
Illegal NgoMIV site found at 880
Illegal NgoMIV site found at 1621
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI.rc site found at 2715

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K2332040 short</partinfo>
-This construct displays GFP fused with SpyTag on the cell surface of E. coli cells under the control of our blue light inducible promoter. Intimin' is a truncated version of Intimin (EaeA intimin), an outer membrane surface protein, which has shown to display proteins of similar size to GFP fused to its N terminal. For blue-light transcriptional induction, cells must also express EL222 ([https://parts.igem.org/Part:BBa_K2332004 BBa_K2332004]).
+This construct displays SpyTag fused to GFP on the cell surface of E. coli cells 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]). Upon blue-light (465nm) exposure, EL222 dimerises and binds to its DNA-binding region within Pblind to recruit RNAP and induce transcription of Intimin'-GFP-SpyTag.
+Intimin' is a truncated version of Intimin (EaeA intimin), an outer membrane surface protein, which has shown to display proteins of similar size to GFP fused to its N terminal.
 This construct allows to directly observe through fluorescence microscopy the functionality of Intimin for cell surface display under blue light induction.