Split-miniSOG for detecting and localizing intracellular protein-protein interactions: application to correlated light and electron microscopy
Files
Accepted manuscript
Date
2019-10-17
Authors
Boassa, Daniela
Lemieux, Sakina P.
Lev-Ram, Varda
Hu, Junru
Xiong, Qing
Phan, Sebastian
Mackey, Mason
Ramachandra, Ranjan
Peace, Ryan E.
Adams, Stephen R.
Version
Accepted manuscript
Embargo Date
Indefinite
OA Version
Citation
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
Abstract
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.
Description
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