Part:BBa_K1899005

J23101-B0032-TetR-B1006- phlFp -GFP

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

Results

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.

The fold change between pSB3K3-phlFp - BBa_E0240(A), negative control (pSB3K3-BBa_E0240) and that of pSB3K3-BBa_J23101-B0032-C0040-B1006- phlFp -E0240(D) is around 13.2 times and 7.01 times respectively. Mean and standard error mean (SEM) are calculated and represented by the height of the bar and the error bar respectively.

This is 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 OD 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.
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