Difference between revisions of "Part:BBa K1761003:Experience"
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mNeonGreen was used to characterise the alx promoter and riboswitch [https://parts.igem.org/Part:BBa_K2348000 BBa_K2348000]. Therefore, we created the working construct [https://parts.igem.org/Part:BBa_K2348012 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 used to characterise the alx promoter and riboswitch [https://parts.igem.org/Part:BBa_K2348000 BBa_K2348000]. Therefore, we created the working construct [https://parts.igem.org/Part:BBa_K2348012 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. mNeonGreen worked well but it must be noted that cells on their owned seem to show emission at about 520 nm. | mNeonGreen was measured with a plate reader with excitation 490 nm, emission 520 nm. mNeonGreen worked well but it must be noted that cells on their owned seem to show emission at about 520 nm. | ||
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| + | == Application TU Eindhoven 2023 == | ||
| + | This part was fused to a long Elastin-Like Polypeptide sequence and used in FRAP experiments. The protein was used in the new composite part [https://parts.igem.org/Part:BBa_K4905016] It was used to characterize part [https://parts.igem.org/Part:BBa_K4905006]. | ||
===User Reviews=== | ===User Reviews=== | ||
Latest revision as of 09:34, 11 October 2023
This experience page is provided so that any user may enter their experience using this part.
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how you used this part and how it worked out.
Applications of BBa_K1761003
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 characterise the alx promoter and riboswitch BBa_K2348000. Therefore, 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. mNeonGreen worked well but it must be noted that cells on their owned seem to show emission at about 520 nm.
Application TU Eindhoven 2023
This part was fused to a long Elastin-Like Polypeptide sequence and used in FRAP experiments. The protein was used in the new composite part [4] It was used to characterize part [5].
User Reviews
UNIQf9cb5ad3aa39e11f-partinfo-00000000-QINU UNIQf9cb5ad3aa39e11f-partinfo-00000001-QINU