Workflow to Select Functional Promoter DNA Baits and Screen Arrayed Gene Libraries in Yeast.

ABSTRACT: The yeast one-hybrid system (Y1H) is used extensively to identify DNA-protein interactions. The generation of large collections of open reading frames (ORFs) to be used as prey in screenings is not a bottleneck nowadays and can be carried out in-house or offered as a service by companies. However, the straightforward use of full gene promoters as baits to identify interacting proteins undermines the accuracy and sensitivity of the assay, especially in the case of multicellular eukaryotes. Therefore, it is paramount to implement procedures for efficient identification of suitable promoter fragments compatible with the Y1H assay. Here, we describe a workflow to identify biologically relevant conserved promoter fragments of Arabidopsis thaliana through simple and robust phylogenetic analyses.

SUBMITTER: Fananas-Pueyo I 

PROVIDER: S-EPMC11580771 | biostudies-literature | 2024 Nov

REPOSITORIES: biostudies-literature