Part:BBa_K1529321

Prhl(RR)_GFP

Prhl(RR) is a promoter that is activated by C4HSL.
We improved Prhl (BBa_R0071) by changing LuxR binding site of Plux (BBa_R0062) to RhlR binding site. (Fig.1)
The bottom structures of LuxR and RhlR are similar to each other, so RhlR can bind to Lux Box. (Fig.2)

On the downstream of the promoter, GFP is inserted as a reporter.

Fig. 1. Our newly designed promoter

Fig. 2. Difference between LuxR and RhlR

To characterize the function of the rhl(RR) promoter (BBa_K1529320), we constructed this part Prhl(RR)_GFP (BBa_K1529321) by inserting rhl(RR) promoter upstream of a GFP coding sequence.
By using reporter cells that contain Prhl(RR)-GFP, we measured the fluorescence intensity of the cells induced by C4HSL (Fig. 2).
We saw that our new rhl(RR) promoter was actually activated by C4HSL through an induction assay (Fig. 3).

Fig. 3. Fluorescence intensity detected by flow cytometer

For more information, see [http://2013.igem.org/Team:Tokyo_Tech/Experiment/RM-lac_Hybrid_Promoter_Assay our work in Tokyo_Tech 2013 wiki].

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 90
  Illegal BamHI site found at 78
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 777