Difference between revisions of "Part:BBa K2332052"
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<partinfo>BBa_K2332052 short</partinfo> | <partinfo>BBa_K2332052 short</partinfo> | ||
− | This construct displays GFP fused with SpyTag on the cell surface of E. coli cells constitutively. Intimin' is a truncated version of Intimin (EaeA intimin) 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. | + | This construct displays GFP fused with SpyTag on the cell surface of E. coli cells constitutively. Intimin' (159kDa) is a truncated version of Intimin (EaeA intimin) 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. It can be also used in combination with cells expressing our SpyCatcher 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. |
Latest revision as of 23:38, 22 October 2017
Intimin'-GFP-SpyTag (constitutive)
This construct displays GFP fused with SpyTag on the cell surface of E. coli cells constitutively. Intimin' (159kDa) is a truncated version of Intimin (EaeA intimin) 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. It can be also used in combination with cells expressing our SpyCatcher 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
- 10INCOMPATIBLE WITH RFC[10]Illegal SpeI site found at 2826
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30
Illegal SpeI site found at 2826 - 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal SpeI site found at 2826
- 25INCOMPATIBLE WITH RFC[25]Illegal SpeI site found at 2826
Illegal NgoMIV site found at 862
Illegal NgoMIV site found at 1603 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 2697