Bacillus subtilis 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 (BBa_K1351010) as well as the sortase YhcS itself (this part), whose overexpression increases the efficiency of the surface display.
This part was generated in RFC10 with the following prefix and suffix:
|prefix with EcoRI, NotI, XbaI, SD and NgoMIV:||GAATTCGCGGCCGCTTCTAGAGTAAGGAGGAGCCGGC|
|suffix with SpeI, NotI and PstI:||TACTAGTAGCGGCCGCTGCAG|
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 in orange. Shine-Dalgarno sequence and stop codons are underlined.
Sequence and Features
- 10COMPATIBLE WITH RFC
- 12COMPATIBLE WITH RFC
- 21COMPATIBLE WITH RFC
- 23COMPATIBLE WITH RFC
- 25Illegal NgoMIV site found at 34
- 1000Illegal BsaI.rc site found at 199
Illegal SapI.rc site found at 508