Part:BBa_K104001
Sensor for small peptide Subtilin
This part is for use in a Bacillus subtilis chassis. It encodes a two-component system that can be used to sense the level of a small antibiotic peptide called subtilin. It belongs to the signalling series of parts (it is a receiver) and can be used to sense quorum sensing peptides from Gram positive organisms. In the [http://2008.igem.org/Team:Newcastle_University Newcastle 2008 iGEM entry] it was used a 'proof of concept' brick to test whether a quorum sensing peptide produced by one species or stain of bacteria could be sensed by a different strain or species.
The part contains two ORFs arranged in an operon, SpaK and SpaR, that are driven by their native RBSs and the promoter PspaRK. Downstream of the SpaRK operon is terminator followed by a RBS and a promoter PspaS.
In response to subtilin, the sensor kinase SpaK will phosphorylate, and hence activate, the response regulator SpaR. The phosphorylated SpaR protein activates the PspaS promoter driving the expression of any downstream genes. In this part there are no ORFs downstream of PspaS but the part has been tested with GFP and mCherry by the Newcastle 2008 team.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
In order to guarantee the expression of GFP, we ran the SDS-PAGE later. Compared with the control group, the WB600 cultured with 0.1mg/ml subtilisin extended significantly strengthened expression of GFP.
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Receiver
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