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A Semiquantitative Protein-Fragment Complementation Assay to Study Protein-Protein Interactions of the Polymerase Complex in Cellula.

Joanna BrunelÉrica UrzuaDenis GerlierLouis-Marie Bloyet
Published in: Methods in molecular biology (Clifton, N.J.) (2024)
Protein-fragment complementation assays (PCAs) are powerful tools to investigate protein-protein interactions in a cellular context. These are especially useful to study unstable proteins and weak interactions that may not resist protein isolation or purification. The PCA based on the reconstitution of the Gaussia princeps luciferase (split-luc) is a sensitive approach allowing the mapping of protein-protein interactions and the semiquantitative measurement of binding affinity. Here, we describe the split-luc protocol we used to map the viral interactome of measles virus polymerase complex.
Keyphrases
  • protein protein
  • binding protein
  • high throughput
  • randomized controlled trial
  • amino acid
  • transcription factor
  • high density
  • dna binding