Part:BBa_K3165001

CcaR v 2.0

CcaR produced naturally by the cyanobacterium Synechocystis sp.PCC6803 acts as a response regulator for the pCpcG2 promoter under the green light-inducible CcaSR system. In its phosphorylated form, CcaR activates the pCpcg2 promoter while the unphosphorylated form retards the activation of the same.

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]

Biology

The CcaR protein acts as a cognate response regulator of the CpcG2 gene (under pCpcG2 promoter). In association with the multi-domain CcaS protein complex CcaR constitutes the green light-inducible CcaSR system. On illumination with green light, the CcaS protein gets autophosphorylated followed by phosphotransfer to CcaR through the histidine kinase domain. The phosphorylated form of CcaR activates the pCpcG2 promoter upregulating the expression of the gene downstream to the promoter. When acted upon by red light, the CcaR protein gets unphosphorylated, preventing the expression of the gene under the pCpcG2 promoter.

Usage

CcaR can be incorporated into a construct along with the CcaS system to replicate the green light-inducible CcaSR system. This part can also be used to characterise the functioning of the CcaSR system in relation to the pCpcG2 promoter at various frequencies and intensities of incident radiation.

References :

(1) Abe, K., Miyake, K., Nakamura, M., Kojima, K., Ferri, S., Ikebukuro, K., & Sode, K. (2013). Engineering of a green-light inducible gene expression system in Synechocystis sp. PCC6803. Microbial Biotechnology, 7(2), 177–183. doi:10.1111/1751-7915.12098
(2) Olson EJ, Hartsough LA, Landry BP, Shroff R, Tabor JJ Characterizing bacterial gene circuit dynamics with optically programmed gene expression signals. Nat Methods. 2014 Mar 9. doi: 10.1038/nmeth.2884. PubMed ID: 24608181