Part:BBa_K2627002

Advanced CcaS with PAS domain knocked out

CcaS belongs to the CcaS-CcaR two-component system, which is a sensor for green light and red light derived from cyanobacterium Synechocystis sp.. CcaS is a sensor protein located on the cell membrane, when it senses the existence of green light or red light, it will autophosphorylate and make the other part of this two-component system--CcaR phosphorylate, thus CcaR can bind to a specific area of DNA, and activate the expression of downstream gene. When green light exists, the gene will be activated while with the existence of red light, the expression will be repressed. Different from the original CcaS derived from Synechocystis sp., we made an improvement of deleting PAS domain on CcaS, making it more sensitive to light, so the expression under green light will be enhanced and the leakage under red light will decrease. In our project, we applied this improved system to increase the sensitivity of the switch as well as the production of PHB.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 606
  Illegal NheI site found at 663
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal XhoI site found at 1379
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Results

Figure 1:Characterization of part BBa_K2627002 and BBa_K592001. Part BBa_K2627002 shows a higher expression index under green light condition and a similar leak index under red light and dark condition.