Part:BBa_K2721024:Design
RFP with SdyTag and SpyCatcher
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 784
Illegal AgeI site found at 600
Illegal AgeI site found at 712 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
The tag-catcher pairs are ideal for binding, immobilization and creating new kinds of protein architectures. Our team use them as scaffold of protein assembly. Here RFP is a reporter.
We have mutated some illegal enzyme sites through synonymous substitutions to make it compatible with RFC[10].
Source
SpyCatcher is derived from Streptococcus pyogenes fibronectin-binding protein. SdyTag is derived from a related fibronectin-binding protein in Streptococcus dysgalactiae.
References
1. Zakeri B, Fierer JO, Celik E, Chittock EC, Schwarz-Linek U, Moy VT, Howarth M. Peptide tag forming a rapid covalent bond to a protein, through engineering a bacterial adhesin. Proc Natl Acad Sci U S A. 2012 Mar 20;109(12):E690-7.
2. Tan LL, Hoon SS, Wong FT. Kinetic Controlled Tag-Catcher Interactions for Directed Covalent Protein Assembly. PLoS ONE 11(10): e0165074. https://doi.org/10.1371/journal.pone.0165074
3. P Hengen. Purification of His-Tag fusion proteins from Escherichia coli. Trends in Biochemical Sciences, 1995,20(7): 285-286.