Uriel Urquiza-Garcia
@uurquiza.bsky.social
Interested in programming plants with DNA, currently building a Plant Artificial Chromosome and Plant Synthetic Genomics developer|CEPLAS
https://www.synthetic-biology.hhu.de/research/urquiza-group
https://www.synthetic-biology.hhu.de/research/urquiza-group
@andrewmillar.bsky.social provided a very concise thread with broader vision in his thread fediscience.org/@A_J_Millar/... and for others to follow up
Andrew Millar (@[email protected])
1/8 🌱🔬 Exciting developments in biology!
Our research investigates the 24-hour, biological clock in plants, now using absolute protein quantification thanks to Uriel Urquiza’s hard work with me, Kare...
fediscience.org
February 22, 2025 at 11:48 AM
@andrewmillar.bsky.social provided a very concise thread with broader vision in his thread fediscience.org/@A_J_Millar/... and for others to follow up
Then joined Andrew and Karen Halliday for dinner down the road at their place as they hosted Mark Stitt and his group form MPI. Though before I discussed just to Andrew that we had done it we got the numbers. The rest well it is history …
February 22, 2025 at 11:48 AM
Then joined Andrew and Karen Halliday for dinner down the road at their place as they hosted Mark Stitt and his group form MPI. Though before I discussed just to Andrew that we had done it we got the numbers. The rest well it is history …
I did plate reader measurements for precise quantities and but also decided to fire up the Millar Hamamatsu EMCCD OrcaII cameras cooled down to -70 Celsius to take an image of the plates that showed the circadian pattern synopsis figure show PRR7
February 22, 2025 at 11:48 AM
I did plate reader measurements for precise quantities and but also decided to fire up the Millar Hamamatsu EMCCD OrcaII cameras cooled down to -70 Celsius to take an image of the plates that showed the circadian pattern synopsis figure show PRR7
Then 2 months before my hard PhD deadline finally after two rounds or circadian sampling 3 days every two hours while purifying MBP-NanoLUC-3Flag-10his standard for the freshest possible enzyme. Adding protease inhibitors and solo working with liquid nitrogen, absolute units protein profiles emerged
February 22, 2025 at 11:48 AM
Then 2 months before my hard PhD deadline finally after two rounds or circadian sampling 3 days every two hours while purifying MBP-NanoLUC-3Flag-10his standard for the freshest possible enzyme. Adding protease inhibitors and solo working with liquid nitrogen, absolute units protein profiles emerged
Without any bench fees I could not afford to waste Furomazine for NanoLUC assays until the full circadian tome series so I was in the blind weather the fusions were fully working apart from some preliminary experiments in protoplasts that indicated correct circadian phase in transient transformation
February 22, 2025 at 11:48 AM
Without any bench fees I could not afford to waste Furomazine for NanoLUC assays until the full circadian tome series so I was in the blind weather the fusions were fully working apart from some preliminary experiments in protoplasts that indicated correct circadian phase in transient transformation
The models informed the selection of lines for creating new reporters using NanoLUC. Went then to rescue mutant lines for CCA1, LHY, PRR7, TOC1 and LUX. Aiming for at least 100 primary T1 events for each line which reduced to around 20 and were then used for a phenotyping FLUC rhythms.
February 22, 2025 at 11:48 AM
The models informed the selection of lines for creating new reporters using NanoLUC. Went then to rescue mutant lines for CCA1, LHY, PRR7, TOC1 and LUX. Aiming for at least 100 primary T1 events for each line which reduced to around 20 and were then used for a phenotyping FLUC rhythms.
I managed to convince Promega for some free reagents for testing NanoLuc in plants with Andrew’s lab instruments which resulted resulted in expertise to work with it in plants plantmethods.biomedcentral.com/articles/10....
Expanding the bioluminescent reporter toolkit for plant science with NanoLUC - Plant Methods
Background Protein data over circadian time scale is scarce for clock transcription factors. Further work in this direction is required for refining quantitative clock models. However, gathering highl...
plantmethods.biomedcentral.com
February 22, 2025 at 11:48 AM
I managed to convince Promega for some free reagents for testing NanoLuc in plants with Andrew’s lab instruments which resulted resulted in expertise to work with it in plants plantmethods.biomedcentral.com/articles/10....
The second path heavily experimental was attempt to quantify the amount of protein. Although tagged lines existed the quantifications that could be derived from this lines seemed impossible. Andrew’s expertise pointed to the use of a new reporter luciferase form @promegacorporation.bsky.social
February 22, 2025 at 11:48 AM
The second path heavily experimental was attempt to quantify the amount of protein. Although tagged lines existed the quantifications that could be derived from this lines seemed impossible. Andrew’s expertise pointed to the use of a new reporter luciferase form @promegacorporation.bsky.social