InterATOME, the Y2H screen service of the OGE team

Protein-protein interactions are critical elements of biological systems and their analysis can provide valuable hints about functions of proteins.

To date, the yeast 2 hybrid (Y2H) system is the only method to probe protein-protein interactions routinely in vivo and with a high-throughput.

The OGE team fully automated and improved the Y2H protocol which was previously used for the AIM-project. Using 384-well plates, successive culture replication in liquid media and robots, the SPOmics-Interactomic Platform is able to screen a pool of 50 DB-X hybrid proteins against the AIM AD-library (about 12000 Arabidopsis proteins) within two months.

OGE Y2H Pipeline

Briefly, ORFs-baits provided by our collaborators directly into the pDEST-DB vector  previously supplied, are utilized for yeast transformation and DB-X hybrid proteins are tested for autoactivator phenotype. DB-X hybrid protein expressing yeasts are then individually grown in selective media, combined into pools of 50 clones and screened against the AIM AD-library by mating.

After identification of candidate interacting AD-proteins, a matricial Y2H assay is performed by de-pooling of the 50 DB-preys and testing against individual candidate AD-baits. A final sequencing step allows unambiguous identifications of both AD- and DB- interacting proteins.  

Every new screened ORF is integrated into the AD-library allowing a continuous network implementation.


Cost (academic lab): 12,500 euros for a screening of up to 50 DB-X hybrid proteins.


dario.monachello @