Abstract
To elucidate the signal cascade that amplifies the nutrient signal and regulates plant growth, we are trying to establish a high throughput screening system to identify novel proteins that interact with the known regulatory factor based on protoplast transient assay. In plant cells, multiple signaling pathways build a complex network to response to the environmental and developmental signals. Consequently, extracting one signaling pathway from such a network can be difficult. In this purpose, to identify the protein interaction is a certain approach, since the signal cascade results from the interactions between signaling molecules.
A regulatory factor fused with the artificial tag is expressed in Arabidopsis protoplast transiently, and the complex including the factor is immunoprecipitated by using the antibody to the tag. The composition of the precipitated fraction is solved by LC/ MS/ MS analysis. This high throughput system should be a useful tool to elucidate a signal cascade efficiently
