A Novel Co-immunoprecipitation Protocol Based on Protoplast Transient Gene Expression for Studying Protein–protein Interactions in Rice

Abstract

Protein–protein interactions constitute the regulatory network that coordinates diverse cellular functions. Co-immunoprecipitation (co-IP) is a widely used and effective technique to study protein–protein interactions in living cells. However, the time and cost for the preparation of a highly specific antibody is the major disadvantage associated with this technique. In the present study, a co-IP system was developed to detect protein–protein interactions based on an improved protoplast transient expression system by using commercially available antibodies. This co-IP system eliminates the need for specific antibody preparation and transgenic plant production. Leaf sheaths of rice green seedlings were used for the protoplast transient expression system which demonstrated high transformation and co-transformation efficiencies of plasmids. The transient expression system developed by this study is suitable for subcellular localization and protein detection. This work provides a rapid, reliable, and cost-effective system to study transient gene expression, protein subcellular localization, and characterization of protein–protein interactions in vivo.