Part:BBa_K2974101:Experience

Medium Promoter (BBa_J23106) Toehold GFP
The construction of the toehold was confirmed through successful transformation in Dh5a cells and sequencing results. The function of the toehold was confirmed through a dual plasmid transformation in BL21 E. coli with the complementary trigger. The dual plasmid transformation presented greater fluorescence measured in light intensity (lux) values in comparison to the single switch transformation, confirming the function of the toehold. Additionally, there were lower lux values associated with the single switch transformation and dual plasmid transformation of the medium-strength promoter toehold when compared to the same toehold switch construct with a strong promoter. The dual plasmid transformation would not succeed without the correct proof of concept toehold intermediate, so the function of this part (BBa_K2974500) has been validated through experimentation and sequencing. eGFP expression in the toehold system for promoters J23100 and J23106 was obtained through successful transformation and light intensity (lux) values were recorded.

Figure 1. Lux values recorded with FluoroCents of the strains of plain LB, plain E. coli cells, trigger-pSB6A1, toehold-pSB3C5, and dual plasmid transformation for promoters BBa_J23100 and BBa_J23106.

Figure 2. Comparison of lux values between promoters BBa_J23100 and BBa_J23106 in strains of plain LB, plain E.coli cells, trigger-pSB6A1, toehold-pSB3C5, and dual plasmid transformation.

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