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Applications of BBa_K3577001

Followed experience is supplemented by 2021 iGEM YiYe-China

Based on K3577001, plasmid pBS1C3 contain trigger was constructed, it is transformed into competent cell through induced expression then observe the fluorescence singnal to detect the degree of gene methylation.

Methods

Plasmid pBS1C3 with toehold-mcherry and pCOLAD-trigger were transformed into E. coliBL21(DE3) strain.Single colony was selected to inoculate LB broth containing kanamycin and cultured overnight.

Results

As shown in the figure 1, the expression of fluorescent protein can be observe in E.coli.And the fluorescence value of only toehold is much lower than that of toehold with trigger, which shows that only toehold can not cause fluorescence,but with trigger lead to a very different fluorescence value（Fig.2). Prove that the toehold system can work to open the hairpin cause a fluorescence.

Conclusion

According to our experimental results：

1. Compare with the control group, there is obvious red light, and the fluorescence value is much higher than others, which is consistent with the initial results of this part.

2. Only very low fluorescence values occur in toehold alone，due to a small amount of leakage,a few fluorescent signals is generated however, it has been shown that the fluorescence value of toehold and trigger is very different from the toehold alone, and the value is much higher than the toehold alone, which proves that its toehold system is working properly.

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