Difference between revisions of "Part:BBa K1351010"
m |
|||
| Line 1: | Line 1: | ||
| − | |||
__NOTOC__ | __NOTOC__ | ||
<partinfo>BBa_K1351010 short</partinfo> | <partinfo>BBa_K1351010 short</partinfo> | ||
| Line 17: | Line 16: | ||
|- | |- | ||
|suffix with AgeI, SpeI, NotI and PstI: | |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> | + | |<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>CGGCCG</span><span style="color:blue">CTGCAG</span> |
|} | |} | ||
Sites of restriction enzymes generating compatible overhangs have the same color: | Sites of restriction enzymes generating compatible overhangs have the same color: | ||
Latest revision as of 15:51, 5 October 2014
Cell wall-anchoring domain of B. subtilis endonuclease YhcR
Cell wall-anchoring domain (amino acids 1086 to 1217) 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.
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 (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[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 81
Illegal BamHI site found at 96 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 52