Experiments | Paris_Bettencourt

A Simple Resistance - dpB.010 -Marionette strain

Abstract

Here, we characterised the expression of our Processor plasmid with the Transcriptional unit dpB.010 [PCin - mScarlet].

We showed that variation of the concentration of pCin’s inducer OCH14 increase the expression level of the construct when mesured at the plate reader

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Materials and methods

We grew E. coli Wild Marionette [1] transformed with the dpB.010-V35 construct to saturation during an overnight culture in LB supplemented with Kanamycin.

The following day, we diluted the culture into fresh media to a 1:100 ratio in a 96 Well plate

Dose response experiment of varying concentration of OHC14(from 1nM to 10 µM) was conducted on the Tecan Infinite® 200 PRO plate reader.

Experiments were done in triplicate

Results

Figure 1 show that varying concentration of OHC14 yielded a range of final fluorescence values consistent with our expectations as Fluorescence increase with concentration

These results validate the functioning of the Processor Transcriptional unit

References

Meyer, A.J. et al. (2019) ‘Escherichia coli “Marionette” strains with 12 highly optimized small-molecule sensors’, Nature Chemical Biology, 15(2), pp. 196–204. Available at: https://doi.org/10.1038/s41589-018-0168-3.