Part:BBa_K861173

mRFP generator controlled by PcstA (glucose-repressible promoter)

RFP is controlled by promoter PcstA, which can be activated by CRP-cAMP complex. Therefore RFP will be expressed at low glucose concentration.

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 712
  Illegal AgeI site found at 824
  
• 1000
  COMPATIBLE WITH RFC[1000]

Vilnius-Lithuania 2019 iGEM team contribution

Vilnius-Lithuania 2019 iGEM team measured measured growth kinetics with varying glucose (5%, 2%, 1% 0.4%, 0.1%, 0.01%, 0.001%) and glycerol (0.4%) concentrations. The measurements were done in a microplate reader, points were taken every 30 minutes for 14 hours and were able then to plot mRFP fluorescence divided by OD 600 against time. In the case of this part characterization, knowing the change in output by varying the glucose concentration allows the part to be used in controlling the protein of interest level in the cell. Full protocol and description can be found here.

Figure 1.The graph showing change in mRFP expression in time with different glucose concentrations (5%, 2%, 1% 0.4%, 0.1%, 0.01%, 0.001%) and glycerol (0.4%). A 9 fold difference can be seen between bacetria grown in 5% and 0% glucose media. The measuring was conducted in 37 ֯C, M9-CA media, E. coli DH5α strain.