Part:BBa_K5335008
SpyCatcher-Linker-Cry6Aa2-Linker-CPPs+SpyTag-6*His
SpyCatcher and SpyTag are able to connect via isopeptides, so Cry6Aa2 will load onto VLP (virus-like particles) through these two connector proteins and act upon the release of VLP. Cell Penetrating Peptides (CPPs) fuse to the C-terminal of Cry6Aa and help the Cry6Aa2 protein enter the cells of nematode worms to trigger the necrosis pathway
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 1517
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30
Illegal PstI site found at 1517 - 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 1622
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 1517
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 1517
Illegal AgeI site found at 692 - 1000COMPATIBLE WITH RFC[1000]
Usage and Biology
The SpyCatcher-SpyTag system was modified from the CnaB2 domain of Streptococcus pyogenes fibronectin binding protein. SpyCatcher is an immunoglobulin-like protein with a molecular weight of about 12 kDa, and SpyTag is a short peptide composed of 13 residues that can recognize each other with high affinity and spontaneously form isopeptide bonds.Cell Penetrating Peptides (CPPs), also known as Trojan peptides, are a family of short hydrophobic peptides that can pass through the cell membrane. A variety of cellular transmembrane peptides have been found to facilitate the uptake of various molecules by cells, such as RNA/DNA, peptides, radioisotopes, liposomes, nanoparticles, and protein molecules.In our project, we fused Cry6Aa2 with SpyCatcher and CPPs, attaching it to virus-like particles with SpyTag, releasing it into the soil environment through the cleavage of engineered bacteria, and CPPs helped our insecticidal proteins enter the nematode body for killing.
Figure . SpyCatcher-Cry6Aa2-CPPs+SpyTag-6*His(pET28a)plasmid vector
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