Part:BBa_K786010

Gene expression for sensory rhodopsin I & II (SRI & SRII)

This part contains the otnpF promoter, after the stimulation of light, Sensory Rhodopsins are able to control the the autophosphorylation activity of CheA. While CheA can phosphorylate OmpR and phosphorylated OmpR is able to stimulate transcription from the otnpF promoter (part R0083).Must be used in E.coli deficient in EnvZ.

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
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 659
  Illegal AgeI site found at 771
  
• 1000
  COMPATIBLE WITH RFC[1000]

Background

The gene expression system

Other than controlling cellular movement by SR, we have also designed a gene expression system induced by SRs.

As explained previously, after light stimulation, SRs can control the autophosphorylation activity of CheA. While CheA can phosphorylate OmpR and phosphorylated OmpR in turn stimulates transcription from the otnpF promoter (part BBa_R0083) [1], we therefore built the construct BBa_K786010. Together with SR sensory systems, gene expression downstream of otnpF promoter can be controlled by different light source.

 

BBa_K786010

Co-transformation of BBa_K786010 with BBa_K786001

With the stimulation of blue light, fusion protein SRII-HtrII-Tsr would increase the autophosphorylation of kinase CheA, thus phosphorylating OmpR and activating downstream genes of R0083.

Co-transformation of BBa_K786010 with BBa_K786002

With the stimulation of blue light, fusion protein SRII-HtrII-Tar would switch off CheA and thus suppress gene expression downstream of R0083.

 

Reference

[1] Igo MM, Ninfa AJ, Stock JB, Silhavy TJ (1989). Phosphorylation and dephosphorylation of a bacterial transcriptional activator by a transmembrane receptor. Genes Dev. 3: 1725-1734.