Difference between revisions of "Part:BBa K1682013"
Stephanieyiu (Talk | contribs) |
|||
Line 1: | Line 1: | ||
− | <partinfo>BBa_K1682013</partinfo> | + | <partinfo>BBa_K1682013 short</partinfo> |
− | <i>P<sub>phoA</sub></i> | + | <i>P<sub>phoA</sub></i> phosphate responsive promoter with GFP translational unitl and terminator BBa_I13504 |
===Biology of <i>P<sub>phoA</sub></i>=== | ===Biology of <i>P<sub>phoA</sub></i>=== |
Revision as of 04:02, 19 September 2015
PphoA - I13504
PphoA phosphate responsive promoter with GFP translational unitl and terminator BBa_I13504
Biology of PphoA
E. coli has multiple native phosphorus sensing and regulation systems that we could use in the construct. Among them, we chose the PhoR/PhoB two-component system (TCS). It contains a sensory histidine kinase PhoR and a partner DNA-binding response regulator PhoB. PhoR is activated under low phosphate concentration, which will then phosphorylate PhoB. The phospho-PhoB is then capable of activating expression of the Pho regulon genes, two of the examples are phoA and phoBR. In high phosphate concentration, phoR is turned into an inhibitory state, which interferes with phosphorylation of PhoB. PhoB is, thus, not capable of activating expression of phoA and phoBR.
Constructs for characterization
With the phosphate (pho) regulon from E. coli, it can be utilized for detecting phosphate level. To make a phospahte-sensing device, we obtained the promoter, PphoA, and combined it with a GFP reporter, BBa_E0240, in BioBrick RFC10 standard so that the promoter activity in different phosphate level can be detected and characterized.
RFU measurement
As shown in Figure 3, PphoA is induced under phosphate limitation and repressed under high phosphate concentration. The fluorescence intensity dropped by 2.99 folds between 0 to 300 μM concentration of phosphate. Furthermore, a plateau is observed starting from the 300 μM phosphate concentration point, suggesting that the dynamic range of PphoA is from 0-300 μM of phosphate.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 755