Improved validation of protein interactions using bicistronic BiFC (Bi2FC).
Physiol Mol Biol Plants
; 30(7): 1047-1054, 2024 Jul.
Article
em En
| MEDLINE
| ID: mdl-39100877
ABSTRACT
Refolding based Bimolecular Fluorescence Complementation (BiFC) has emerged as an important in vivo technique to identify protein interactions. Significant improvements have been made to enhance the detection capacities of BiFC, however less attention has been paid to the detection of expression levels of proteins. Here we demonstrate development and validation of an improved method to identify protein interactions that incorporates an expression control based on bicistronic expression of the protein of interest and a fluorescent protein separated by a self-cleaving peptide. This method gives robust identification of positive interactions and more reliably identifies absence of interactions. We also show an earlier identified non-interacting pair in yeast two-hybrid (Y2H) to be interacting in vivo. Supplementary Information The online version contains supplementary material available at 10.1007/s12298-024-01477-y.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Idioma:
En
Revista:
Physiol Mol Biol Plants
Ano de publicação:
2024
Tipo de documento:
Article