Split-miniSOG for detecting and localizing intracellular protein-protein interactions: application to correlated light and electron microscopy
Lemieux, Sakina P.
Peace, Ryan E.
Adams, Stephen R.
Ellisman, Mark H.
Ngo, John T.
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Citation (published version)Daniela Boassa, Sakina Lemieux, Varda Lev-Ram, Junru Hu, Qing Xiong, Sebastian Phan, Mason Mackey, Ranjan Ramachandra, Ryan Peace, Stephen Adams, Mark Ellisman, and John T. Ngo. 2019. "Split-miniSOG for detecting and localizing intracellular protein-protein interactions: application to correlated light and electron microscopy." Cell Chemical Biology, Volume 26, Issue 10, Pages 1407-1416.e5. https://doi.org/10.1016/j.chembiol.2019.07.007
A protein-fragment complementation assay (PCA) for detecting and localizing intracellular protein-protein interactions (PPIs) was built by bisection of miniSOG, a fluorescent flavoprotein derived from the light, oxygen, voltage (LOV)-2 domain of Arabidopsis phototropin. When brought together by interacting proteins, the fragments reconstitute a functional reporter that permits tagged protein complexes to be visualized by fluorescence light microscopy (LM), and then by standard as well as “multicolor” electron microscopy (EM) via the photooxidation of 3-3’-diaminobenzidine (DAB) and its derivatives.
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