Part:BBa_K1899005

J23101-B0032-TetR-B1006- phlFp -GFP

This part is constructed for investigating the effect of TetR over phlFp.

Results

The fold change between construct A (pSB3K3-BBa_phlFp -E0240) and negative control (pSB3K3-BBa_E0240), and that of construct D (pSB3K3-BBa_J23101-B0032-C0040-B1006- phlFp-E0240) is around 13.2 times and 7.01 times respectively.

We hypothesized that this may due to the toxicity of BBa_C0040 (TetR). The toxicity reduces the growth rate of the E. coli containing this plasmid. The strong promoter BBa_J23101 makes this effect more significant when doing the characterisation. Though all the data were collected with OD₆₀₀ within the mid-log range, there is still a variation between them, making a significant difference in the RFU. It is ungrounded to say TetR interferes the functionality of phlFp at this stage.

Fig 1. Comparison on fluorescence expression levels of construct A (BBa_phlFp-E0240) and construct D (BBa_J23101-B0032-C0040-B1006- phlFp -E0240). Negative control represents BBa_E0240. Characterization was done using E. coli strain JW0336. Cells were first precultured overnight and were subcultured to mid-log phase where GFP emission measurements were made using an EnVision® multilabel reader. This result was obtained by combining 3 characterization data obtained in 3 different days. Error bar present SD from 3 biological replicates.

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 7
  Illegal NheI site found at 30
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 1539