Labour‐saving construction of a target protein interaction network by selective culture and high‐throughput sequencing

Abstract Yeast two‐hybrid (Y2H) is a classic method of screening for protein‐protein interactions. However, the operation process is labor intensive and time consuming, and there is a high possibility of false positives and false negatives. Combined with wet lab operation and bioinformatics analysis, we developed a novel method of Y2H library screening using Chrysanthemum morifolium CmMPK3 as an example. The protocol can not only greatly simplify the steps of traditional Y2H library screening but also identify as many interacting proteins as possible. Furthermore, this protocol is applicable to any species, even if no genomic information is available yet.