Difference between revisions of "Part:BBa K1351008"

Revision as of 12:57, 5 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. 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.

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:	ACCGGTTAATACTAGTAGCGGCCGCCTGCAG

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 1: / Line 1: @@
 __NOTOC__
 <partinfo>BBa_K1351008 short</partinfo>
@@ Line 8: / Line 7: @@
 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.
+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:
+|<span style="color:blue">GAATTC</span><span style="color:green">GCGGCCGC</span>T<span style="color:red">TCTAGA</span>GT<u>AAGGAGG</u>A<span style="color:orange">GCCGGC</span>
+|-
+|suffix with AgeI, SpeI, NotI and PstI:
+|<span style="color:orange">ACCGGT</span><u>TAA</u>T<span style="color:red">AC<u>TAG</u><u>T</u></span><u>A</u><span style="color:green"><u>G</u>CGGCCGC</span><span style="color:blue">CTGCAG</span>
+|}
+Sites of restriction enzymes generating compatible overhangs have the same color:
+<span style="color:blue">EcoRI</span> and <span style="color:blue">PstI</span> in blue, <span style="color:green">NotI</span> in green, <span style="color:red">XbaI</span> and <span style="color:red">SpeI</span> in red, <span style="color:orange">NgoMIV</span> and <span style="color:orange">AgeI</span> in orange. Shine-Dalgarno sequence and stop codons are underlined.
 <!-- Add more about the biology of this part here
@@ Line 13: / Line 27: @@
 <!-- -->
-<span class='h3bb'>Sequence and Features</span>
+===Sequence and Features===
 <partinfo>BBa_K1351008 SequenceAndFeatures</partinfo>