Part:BBa_K2023013:Experience
We aimed to asses the XylR production by E.coli cells transformed with the BioBrick BBa_K2023013 composed of Pr-RBS-XylR-RBS-mRFP-Terminator. This BioBrick was designed to monitor XylR synthesis by our cells.
To do so, we thaw a tubes of transformed cells on ice for 10 minutes, we added different volumes of SOC medium to obtain several dilutions (1/2, 1/4 and 1/8), we plated different concentrations of bacteria cells on petri dish with LB Agar + Chloramphenicol at 25µg/L final and incubate overnight.
We obtained the expected results : red colonies were shown on the petri dish (Figure) due to the synthesis of the red protein mRFP meaning that the Pr promoter trigger XylR and mRFP synthesis.
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<center><figcaption>Figure: Monitoring of XylR production though control of the activity of the Pr promoter. Modified bacteria containing XylR coding device with mRFP grown on a petri dish. The red color of the bacteria indicate that mRFP protein is produce and therefore that XylR is produce too.
This construction will not allow XylR quantification but those results show that XylR protein was constitutively produce by our genetically modified bacteria containing XylR coding device with mRFP. We wanted to realize a XylR-mRFP fusion but this part was too big for IDT synthesis so we created the BioBrick BBa_K2023010 (XylR-His)
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