CSEP0174: Yeast two-hybrid next-generation interaction screening (Y2H-NGIS) to discover protein-protein interactions influencing barley powdery mildew disease resistance and susceptibility.

ABSTRACT: The barley powdery mildew effector (CSEP0174) dataset was part of a larger yeast two-hybrid (Y2H) screen to identify novel proteins involved in barley immune responses. Y2H screening was coupled with next-generation sequencing to identify and quantify interacting proteins. Individual baits were mated to a cDNA prey library derived from a 0 - 48 hour time course of infected leaf tissue (1.1 x 107 primary clones). Screens were performed in batch liquid culture to enrich yeast populations for cells expressing positive interactions. After two rounds of enrichment under selective (Histidine absent) and non-selective (Histidine present) conditions, yeast cells were collected. Y2H plasmids were extracted and prey cDNA amplicons were generated via low-cycle PCR. Fragmented amplicons were used as input to generate sequencing libraries and processed on the NovaSeq6000 platform. Reads were mapped to the barley and Blumeria hordei genomes and read counts were analyzed using a custom data processing and scoring pipeline. Putative interactors were cloned and binary Y2H was used to confirm interactions.

ORGANISM(S): Hordeum vulgare

PROVIDER: GSE299565 | GEO | 2026/10/05

REPOSITORIES: GEO