Intermediate

Part:BBa_K1899010

Designed by: iGEM2016_HKUST   Group: iGEM16_Hong_Kong_HKUST   (2016-10-18)


B0032-TetR

TetR is conjugated to a medium strength RBS.

Construct for Characterisation

Fig.1 tetR-tetp with reporter gene.

In order to characterise the efficiency of this construction intermediate, it was ligated with a construct containing tetp and GFP reporter gene, BBa_E0240. The expression of TetR was driven by a strong constitutive promoter, BBa_J23101, and terminated by BBa_B1006 terminator. The assembly processes were performed in BioBrick RFC10 standard.

Results

Experiments on comparing the GFP expression driven by tetp from constructs with and without BBa_B0032-tetR were performed. Negative control used was BBa_E0240. Results indicated that BBa_B0032-tetR driven by BBa_J23101 and terminated by BBa_B1006 could reduce the GFP expression by 3.6 times.

Fig 2. Comparison on the GFP expression of construct with and without BBa_B0032-tetR. BBa_E0240 was used as negative control. Characterisation was done using E. coli strain JW0336. Cells were first pre-cultured 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 characterisation 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
    COMPATIBLE WITH RFC[1000]


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Categories
//chassis/prokaryote/ecoli
Parameters
None