Journal
ANALYTICAL CHEMISTRY
Volume 75, Issue 7, Pages 1584-1589Publisher
AMER CHEMICAL SOC
DOI: 10.1021/ac020731c
Keywords
-
Categories
Funding
- NCI NIH HHS [R01 CA082214, R24 CA092865, R0-1 CA82214, R24 CA92865] Funding Source: Medline
Ask authors/readers for more resources
In this study we developed an inducible synthetic renilla luciferase protein-fragment-assisted complementation-based bioluminescence assay to quantitatively measure real time protein-protein interactions in mammalian cells. We identified suitable sites to generate fragments of N and C portions of the protein that yield significant recovered activity through complementation. We validate complementation-based activation of split synthetic renilla luciferase protein driven by the interaction of two strongly interacting proteins, MyoD and Id, in five different cell lines utilizing transient transfection studies. The expression level of the system was also modulated by tumor necrosis factor alpha through NFkappaB-promoter/enhancer elements used to drive expression of the N portion of synthetic renilla. luciferase reporter gene. This new system should help in studying protein-protein interactions and when used with other split reporters (e.g., split firefly luciferase) should help to monitor different components of an intracellular network.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available