Part:BBa_K3725070:Experience

Experience
The T7 Fusarium Trigger (Part BBa_K3725070) and the Fusarium Toehold w/ GFP Reporter (Part BBa_K3725020) are intended to be compatible with each other and be used in conjunction. To test the compatibility of the trigger sequence with the toehold sequence, a dual-plasmid transformation was performed, and fluorescence was measured in comparison with cells transformed with only the toehold part and pUC19 (positive control). All samples were divided by optical density to obtain a standardized unit. The fluorescence per optical density unit measurement of cells transformed with both the toehold and trigger sequences was not statistically greater in comparison to the measurements of cells transformed with only the toehold part and pUC19 plasmid.

Figure 1: Mean fluorescence/OD of Fusarium dual plasmid transformation compared to toehold and plain LB with SEM error bars. DP stands for dual plasmid, TH stands for toehold only. Ran at a gain of 60.

So, Lambert iGEM decided to use a redesigned toehold switch sequence, Improved Fusarium Toehold w/ GFP Reporter (Part BBa_K3725022), obtained from NUPACK. Consequently, another dual-plasmid transformation was performed using the new switch sequence and the trigger sequence, and fluorescence was measured. The fluorescence per optical density unit measurement of cells transformed with both plasmids was greater and had a statistically significant difference in comparison to the measurements of cells transformed with only the toehold part and pUC19 plasmid.

Figure 2: Mean fluorescence/OD of IPTG-induced Fusarium pair 2 dual plasmid transformation compared to toehold and pUC19 with SEM error bars. Ran at gain of 40.

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