FACS-based in planta BiFC screening

• Applicant: Professor Dr. Klaus Harter
• Subject Area: Organismic Interactions, Chemical Ecology and Microbiomes of Plant Systems ; Plant Cell and Developmental Biology
• Term: from 2007 to 2012
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 38601546

For the first round of the SPP1212, our proposed research was divided into two parts. In the first part we proposed the exploration of the functional role of two-component signalling systems (TCS) in Arabidopsis-microbe interactions. This part was not recommended for funding. In the second part we had proposed to establish a novel Bimolecular Fluorescence Complementation (BiFC)-based in vivo two-hybrid system for the identification and characterisation of protein-protein interactions in living plant cells by using fluorescence activated cell sorting (FACS). The establishment of the in vivo BiFC based interaction screen was more challenging than expected. However we have managed to develop a working protocol. Our approach is of great interest to most groups in and also outside of the SPP1212 that are working on in planta protein-protein interaction and protein complex formation. This proposal is targeted towards further improving the FACS-based in planta BiFC screening approach and offering the technology and expertise to the members of the SPP1212 in a dedicated fashion. Furthermore, we also offer research support, by providing dedicated cytometric and FACS expertise for other applications besides the BiFC screen to the SPP1212 community such as promoter transactivation, sorting of cells and nuclei for “-omics” work (e.g. transcriptomics, proteomics, ChIP-chip) and monitoring cell cycle (e.g. DNA content).

• DFG Programme: Priority Programmes
• Subproject of: SPP 1212: Microbial Reprogramming of Plant Cell Development