Composite

Part:BBa_K1899008

Designed by: iGEM16_Hong_Kong_HKUST   Group: iGEM16_Hong_Kong_HKUST   (2016-10-13)

phlFp-E0240

Construct for characterization

Fig.1 phlFp with reporter gene.

In order to compare the promoter strength of the phlFp with the other two promoters, tetp and lacp, it was ligated with a GFP reporter gene, BBa_E0240, in BioBrick RFC10 standard. With the presence of a green fluorescence protein reporter gene, GFP protein would be produced during the characterization, thus, the strength of phlFp could be compared to that of the other two promoters in our project, lacp and tetp.


Results

Experiments on comparing the strength of phlFp, tetp and lacp, were performed. Results indicated that phlFp is the strongest among the three, with 1.96 times and 6 times stronger than that of tetp and lacp, respectively.

Fig 2. Comparison on the strength of phlFp, tetp and lacp. 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
    COMPATIBLE WITH RFC[12]
  • 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 737


[edit]
Categories
//chassis/prokaryote/ecoli
Parameters
None