Cell wall-anchoring domain of B. subtilis endonuclease YhcR
Cell wall-anchoring domain (amino acids 1086 to 1217) of Bacillus subtilis endonuclease YhcR (UniProt P54602). YhcR is secreted and covalently anchored to the petidoglycan wall by the sortase YhcS.
A YhcR-based surface display system has been shown to covalently anchor heterologous proteins to the B. subtilis cell wall (Liew et. al., 2012). It was used in the BaKillus project to display pathogen-specific peptides and mediate adhesion of B. subtlis to pathogens.
It consists of four components: the sortase substrate with a signal peptide (BBa_K1351008), a linker (BBa_K1351009) and a cell wall-anchoring domain (this part) as well as the sortase YhcS itself (BBa_K1351011), whose overexpression increases the efficiency of the surface display.
This part was generated in a modified version of RFC25, where a strong Shine Dalgarno Sequence (SD) is included, and has the following prefix and suffix:
|prefix with EcoRI, NotI, XbaI, SD and NgoMIV:||GAATTCGCGGCCGCTTCTAGAGTAAGGAGGAGCCGGC|
|suffix with AgeI, SpeI, NotI and PstI:||ACCGGTTAATACTAGTAGCGGCCGCTGCAG|
Sites of restriction enzymes generating compatible overhangs have the same color:
EcoRI and PstI in blue, NotI in green, XbaI and SpeI in red, NgoMIV and AgeI in orange. Shine-Dalgarno sequence and stop codons are underlined.
Sequence and Features
- 10COMPATIBLE WITH RFC
- 12COMPATIBLE WITH RFC
- 21Illegal BamHI site found at 81
Illegal BamHI site found at 96
- 23COMPATIBLE WITH RFC
- 25COMPATIBLE WITH RFC
- 1000Illegal SapI.rc site found at 52