Fluorescence labelling of re-coded E.coli w/ non-canonical chem. entities for single mol. tracking
Автор: GCE4All Research Center
Загружено: 2024-12-03
Просмотров: 258
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Talk given by Filip Ilievski (Magnus Johansson lab, Uppsala University, Sweden) as part of the International GCE Webinar series. Live talk given on November 21st, 2024.
Single molecule tracking (SMT) is a powerful tool for real-time studies of protein interactions in living cells. Self-labelling tags such as Snap- and Halo-tag have revolutionized SMT, due to highly stable, long-lived signal of the orthogonal dye-ligand conjugates. Akin to fluorescent proteins for SMT, their size and structure limits their use in labelling small and dynamically delicate proteins. We introduce FLORENCE – a universal labelling method for SMT, based on genetic code expansion (GCE). We have overcome significant caveats previously related to re-coded strains, vectors and dyes, and, for the first time, report successful tracking of intra-cellular proteins in living E.coli.
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