Difference between revisions of "Part:BBa K2332015:Design"

 
(→‎Design Notes)
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===Design Notes===
 
===Design Notes===
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Amberless E. coli must be used to see the effects of photocageing spyCatcher
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This construct was designed to not contain any other amber stop codon but at position 31 (Lys31X) (Amberless E. coli must be used to see the effects of photocageing spyCatcher).
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We decided to fuse SpyCatcher to the N-terminal of a truncated version of Intimin as this has been previously proven to function effectively for cell surface display of up to 30kDa passenger proteins (Wentzel et al., 2001). We also included a linker between Intimin and SpyCatcher to facilitate mobility of SpyCatcher on the cell surface and a HisTag for protein purification.
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===Source===
 
===Source===

Revision as of 22:38, 22 October 2017


Intimin'-mutSpyCatcher (Lys31X, for photocaging)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 819
    Illegal NgoMIV site found at 1560
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

This construct was designed to not contain any other amber stop codon but at position 31 (Lys31X) (Amberless E. coli must be used to see the effects of photocageing spyCatcher). We decided to fuse SpyCatcher to the N-terminal of a truncated version of Intimin as this has been previously proven to function effectively for cell surface display of up to 30kDa passenger proteins (Wentzel et al., 2001). We also included a linker between Intimin and SpyCatcher to facilitate mobility of SpyCatcher on the cell surface and a HisTag for protein purification.

Source

SpyCatcher: BBa_K1159200

References