Part:BBa_K1761003:Experience

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Application TU Eindhoven 2015

mNeonGreen is used as an intracellulair signaling domain. In combination with the Luciferase NanoLuc a working BRET (Bioluminescence Resonance Energy Transfer) couple is formed. For a broader description on how we used this part, take a look at part BBa_K1761001 [1], because it also contains this part.

Another application of mNeonGreen as signaling domain, is by using it with another fluorophore (for example mTurquoise2) and preform a FRET (Förster Resonance Energy Transfer). We as a team preferred to use BRET, because then substrate needs to be added, instead of using a laser for excitation, which can lead to more background noise.

mNeonGreen was also expressed with the spromotor J23101 [2] for quantification. This promotor was also used by our team for the 2015 InterLab Study [http://2015.igem.org/Team:TU_Eindhoven/Project/Interlab], but then with a GFP protein, part I13504 [3]. Even though no RBS was introduced into the composite part, the cells still showed fluorescence which could be measured.

Application NAWI_Graz 2017

mNeonGreen was used to charakterise the alx promoter and riboswitch BBa_K2348000. Therfore we created the working construct BBa_K2348012 containing the alx promoter and riboswitch, a TEV-site fused with f-degron, mNeonGreen with a Flag-tag and a T7 terminator. mNeonGreen was measured with a plate reader with excitation 490 nm, emission 520 nm.

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