Part:BBa_K3524004

Psal pro-EGFP-T7 ter

The bio-sensor (nahR and Psal) for the detection of salicylic acid we adopted is originally on the chromosome of Pseudomonas Stutzeri. With the appearance of salicylic acid, the protein translated from nahR binds with Psal promoter and initiates the expression of the enzyme that can utilize the acid as one of the energy sources for Pseudomonas Stutzeri. In our design, we replaced the essential gene with the GFP gene to test our framework. With GFP, the situation is that the edited bacteria can be fluorescent with the existence of salicylic acid.

Contribution

To build up a regulable genetic circuit, we chose the bio-sensor (nahR gene and Psal promoter) for the detection of salicylic acid we adopted is originally on the chromosome of Pseudomonas Stutzeri. With the appearance of salicylic acid, the protein translated form nahR binds with PsaI promoter and initiates the expression of the enhancer green fluorescent protein (EGFP).

Figure 1

Our team has verified this bio-sensor worked well. The future iGEM teams can exchange the EGFP to other functional proteins to make some significant utiles. For example, our team choose an essential gene GAPDH, belongs to Tatumella citrea, which can provide phosphorus for crops[1][2] to replace the EGFP . As a result, in the presence of salicylic acid, GADPH can be expressed, and T. citrea can survive. Since salicylic acid is a root exudate, this genetic circuit that T. citrea can commensalism with crops.

Refernce

[1] Whitelaw, M.a. “Growth Promotion of Plants Inoculated with Phosphate-Solubilizing Fungi.” Advances in Agronomy, 1999, pp. 99–151., doi:10.1016/s0065-2113(08)60948-7.

[2] Bar-Yosef, B., et al. “Pseudomonas Cepacia-Mediated Rock Phosphate Solubilization in Kaolinite and Montmorillonite Suspensions.” Soil Science Society of America Journal, vol. 63, no. 6, 1999, pp. 1703–1708., doi:10.2136/sssaj1999.6361703x. 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]