期刊
JOURNAL OF VIROLOGICAL METHODS
卷 176, 期 1-2, 页码 108-111出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jviromet.2011.04.028
关键词
Viral disease; Protein-protein interaction; Split luciferase complementation assay
资金
- NSF/EPSCoR [0091948]
- Center of Excellence in Drought Tolerance through the South Dakota 2010 Initiative
- South Dakota Agri. Exp. Station
- South Dakota 2010 Research Center
- BCAAP (Biological Control and Analysis of Applied Photonics) [3SG163]
- South Dakota Agricultural Experiment Station [3AH203]
- NIAID [A1078177]
Intraviral protein-protein interactions are critical for virus survival in the host. Discovery of such interactions is important to understand molecular mechanisms of viral replication and pathogenesis. The development of a cell-based assay that can be employed to examine systematically viral protein interactions is described. The method, known as the split luciferase complementation assay (SLCA), is based on the principle that N- and C-terminal domains of luciferase alone do not emit luminescence; however, if fused to interacting proteins the two non-functional halves can be brought into close enough proximity through a specific protein-protein interaction to restore the functions of the enzyme and emit detectable light. The well-studied influenza B polymerase acidic protein (PA) and basic protein 1 (PB1) interaction was used as a model system to develop the assay. Consistent with previous studies, a strong PA-PB1 interaction was demonstrated in the assay. The PA-PB1 interaction was also disrupted by single amino acid mutations in the N-terminal domain of PB1 that is responsible for binding PA. The described SLCA is highly specific and easy to perform, and thus may be useful for studying protein-protein interactions in viral diseases. (C) 2011 Elsevier B.V. All rights reserved.
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