Coding

Part:BBa_K5335001

Designed by: Zekun Luo   Group: iGEM24_HZAU-China   (2024-09-27)
Revision as of 08:09, 30 September 2024 by Jerseylandd (Talk | contribs)


The fusion protein of SpyCatcher with EGFP.

Spycatcher can form a covalent connection with Spytag. Using this property, we plan to link Spycatcher with functional proteins so that it can be displayed on the surface of VLP particles in combination with MS2-Spytag.


Usage and Biology

The SpyTag-SpyCatcher system is a fast, reliable and irreversibly linked protein coupling tool developed in recent years, which enables the two to recognize each other with high affinity and spontaneously form isopeptide bonds. Taking advantage of this feature, we used a fusion protein composed of SpyCatcher and EGFP to verify whether it could be displayed on the surface of VLP to confirm the reliability of our delivery platform.

Figure 1. The structure of the connection complex formed by SpyTag and SpyCatcher.
The image is from the PDB database with the PDB ID 4MLS. The cyan part represents the SpyCatcher and the orange part represents the 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 site found at 480
    Illegal BsaI.rc site found at 1187

SpyCatcher-EGFP was constructed on the PUC57 mini plasmid. The upstream MS2-SpyTag is used to verify the binding capability. (Figure 1.)


Figure 1. PUC57 mini plasmid with spycatcher-egfp sequence.


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Parameters
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