Team:Valencia UPV/Improve

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Part Improvement

The BioBrick chosen to be improved was the YFP coding sequence: BBa_K592101. To do this, we first standardized the part to be compatible with the GoldenBraid 3.0 assembly method creating the part BBa_K2656021. Then, we created the part BBa_K2656020. This part is the CDS of the YFP with the addition of the degradation tag ssRA LVA.

To test the effect of the degradation tag, we designed an experiment with which we measured the increase in protein degradation due to this tag. To perform this experiment, we assembled two composite parts with the same promoter, RBS and terminator:

Once the experiment was carried out, the results were plotted and figure 1 was obtained, in which we can observe that the growth of the bacteria with both constructions was very similar, while the fluorescence had a clear variation.

Figure 1. Experimental results of the fluorescence comparison experiment between the transcriptional unit with BBa_K2656021 and the one with BBa_K2656020.

These data were optimized with our model and the parameters from Table 1 were obtained. With these parameters it is possible to obtain that the protein degradation of the protein with the degradation tag is around twice as much as without the tag.

Optimized parameters

Values

Translation rate p

PoI degradation rate dp

Dilution rate μ

CONTACT US igem.upv.2018@gmail.com