Protein Interactions in Live Cells Monitored by β-Galactosidase Complementation
نویسندگان
چکیده
The characterization of protein interactions is important to the understanding of signal transduction pathways and cellular processes. Here we describe a method utilizing β-galactosidase (βgal) complementation that can monitor protein interactions in live mammalian cells (Rossi et al. 1997, 2000; Blakely et al. 2000). In brief, the method involves expressing chimeric proteins consisting of two potentially interacting proteins of interest fused to complementing β-gal deletion mutants. When the two proteins of interest interact, the β-gal mutants complement, reconstituting an active β-gal enzyme. Some of the advantages of β-gal complementation are: (1) It is a direct assay, that is, a signal is generated at the site of the interaction, and activation of a reporter gene is not required; (2) it is a sensitive assay, due to enzymatic amplification of the product; (3) over-
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