An optimized split-ubiquitin cDNA-library screening system to identify novel interactors of the human Frizzled 1 receptor.

ABSTRACT: The yeast split-ubiquitin system has previously been shown to be suitable to detect protein interactions of membrane proteins and of transcription factors in vivo. Therefore, this technology complements the classical split-transcription factor based yeast two-hybrid system (Y2H). Success or failure of the Y2H depends primarily on the ability to avoid false-negative and false-positive hits that become a limiting factor for the value of the system, especially in large scale proteomic analyses. We provide here a systematic assessment of parameters to help improving the quality of split-ubiquitin cDNA-library screenings. We experimentally defined the optimal 5-fluoroorotic acid (5-FOA) concentration as a key parameter to increase the reproducibility of interactions and, at the same time, to ke

SUBMITTER: Dirnberger D 

PROVIDER: S-EPMC2330230 | biostudies-literature | 2008 Apr

REPOSITORIES: biostudies-literature