Types | DnaRegion
|
Roles | Coding
CDS
|
Sequences | BBa_K1351008_sequence (Version 1)
|
Description
Signal peptide (amino acids 1 to 46) of
Bacillus subtilis endonuclease YhcR (UniProt
P54602. YhcR is secreted and covalently anchored to the petidoglycan wall by the sortase YhcS. Despite its signal protein of RR-type, which is typical for proteins exported via the Tat pathway, export of YhcR is not Tat-dependent and seems to function via the Sec pathway (Tjalsma
et. al., 2004).
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 (this part), a linker (
BBa_K1351009) and a cell wall-anchoring domain (
BBa_K1351010) as well as the sortase YhcS itself (
BBa_K1351011), whose overexpression increases the efficiency of the surface display.
Notes
The native signal peptide of YhcR was used instead of known Sec-dependent signal peptides as Liew et. al. did 2012.
Source
This part was generated by amplification from B. subtilis W168 gDNA with the primers listed below, followed by digestion with EcoRI and PstI and ligation into pSB1C3.
SP_YhcR_ ENX_SD_Ngo_fwd: gatcGAATTCgcggccgctTCTAGAgtaaggaggaaGCCGGC ATGCTGTCTGTCGAAATGATAAGC
SP_YhcR_SNP_Age_rev: gatcCTGCAGcggccgctACTAGTattaACCGGT AGCTTCGAACGTGTACATTACATTTAAG