Difference between revisions of "Part:BBa K1351008"

Latest revision as of 16:12, 16 October 2014

Signal peptide of B. subtilis endonuclease YhcR

Signal peptide (amino acids 1 to 46) of Bacillus subtilis endonuclease YhcR (UniProt [http://www.uniprot.org/uniprot/P54602 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 [http://2014.igem.org/Team:LMU-Munich/Project/Bakillus BaKillus project] to display pathogen-specific peptides and mediate adhesion of B. subtilis 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.

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

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

@@ Line 2: / Line 2: @@
 <partinfo>BBa_K1351008 short</partinfo>
-Signal peptide (amino acids 1 to 46) of ''Bacillus subtilis'' endonuclease YhcR (UniProt [http://www.uniprot.org/uniprot/P54602 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).
+Signal peptide (amino acids 1 to 46) of ''Bacillus subtilis'' endonuclease YhcR (UniProt [http://www.uniprot.org/uniprot/P54602 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 [http://2014.igem.org/Team:LMU-Munich/Project/Bakillus BaKillus project] to display pathogen-specific peptides and mediate adhesion of ''B. subtlis'' to pathogens.
+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 [http://2014.igem.org/Team:LMU-Munich/Project/Bakillus BaKillus project] to display pathogen-specific peptides and mediate adhesion of ''B. subtilis'' to pathogens.
 It consists of four components: the sortase substrate with a signal peptide (this part), a linker ([https://parts.igem.org/Part:BBa_K1351009 BBa_K1351009]) and a cell wall-anchoring domain ([https://parts.igem.org/Part:BBa_K1351010 BBa_K1351010]) as well as the sortase YhcS itself ([https://parts.igem.org/Part:BBa_K1351011 BBa_K1351011]), whose overexpression increases the efficiency of the surface display.