Part:BBa_K1351036:Design
Quorum sensing two component system (AIP-II sensing histidine kinase agrC, response regulator agrA)
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 1139
Illegal AgeI site found at 1114 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 475
Design Notes
This part contains the native sequences of agrC-II and agrA from Staphylococcus aureus N315. Since these two parts are forming a two component system, this is actually a composite part. The basic parts agrC-II and agrA can be yielded by using the NgoMIV or AgeI restriction sites with appropiate restriction sites in the pre- or suffix.
Source
This part is derived from Staphylococcus aureus N315 gDNA by PCR using primers with modified RFC25 prefix and suffix:
agrA-fwd: gatcgaattccgcggccgcttctagataaggaggagccggcATGAAAATTTTCATTTGCGAAGACG
agrA-rev: gatcctgcagcggccgctactagtattaaccggtTATTTTTTTAACGTTTCTCACCG
agrC-II-fwd: gatcgaattccgcggccgcttctagataaggaggagccggcTTGATTCCAACTTTTTCATCTATC
agrC-II-rev: gatcctgcagcggccgctactagtattaaccggtGTTGTTAATAATTTCAACTTTTTGAATAAAG
The two intermediated parts agrC-II and agrA were then fused by using the restriction sites EcoRI and SpeI for agrC-II, as well as XbaI and PstI for agrA, and ligation into pSB1C3 (linearized with EcoRI and PstI), thus creating the composite part agrC-IIA.
References
Novick, R. P. (2003). "Autoinduction and signal transduction in the regulation of staphylococcal virulence." Molecular Microbiology 48(6): 1429-1449.
The Staphylococcus aureus N315 gDNA was kindly provided by [http://www.bath.ac.uk/bio-sci/contacts/academics/susanne_gebhard/ Dr. Susanne Gebhard] (University of Bath)