Part:BBa_K3885312
P70a-UTR2-deGFP
Under the combination of P70a promoter and UTR2, green fluorescent protein can be expressed.
Usage and Biology
According to the lecture, we selected three promoters for Cell-Free systems: P70a, P70b and P70c and combined with three Ribosome Binding Sites (RBS) UTR1, UTR2 and UTR3 to express deGFP gene. The fluorescence intensity of deGFP was measured by adding varied concentrations of plasmids into the Cell-Free system. Simultaneously, experimental results and modeling data were compared, and the optimal promoter RBS combination was obtained in order to express deGFP in a Cell-Free system.
Result
Figure 1. Comparing expression in TXTL reactions to the model. A) Maximum rate of deGFP synthesis expressed from varying concentrations of P70a-UTR2-deGFP. B,C,D) Kinetics of deGFP synthesis for the plasmid P70a-UTR2-deGFP at three concentrations: one below, one at, and one above saturation (A) 1 nM, (B) 5 nM, (C) 10 nM, shown for the first 3 hours.
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 64
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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