Three-hybrid systemを用いたABA受容体のスクリーニング

概要

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The yeast three-hybrid system is an extension of the two-hybrid system, and designed to detect the interaction between a small molecule and its receptor. In addition to the two hybrid fusion proteins, the three-hybrid system requires a third hybrid ligand (Bait). One-half of the hybrid ligand consists of dexamethasone. Binding of dexamethasone to the glucocorticoid receptor fused to a DNA-binding domain (Hook) serves to anchor the ligand to the promoter of a reporter gene. Interaction of the tethered orphan ligand, the other half of the hybrid ligand, with a protein receptor fused to an activation domain (Fish) initiates transcription of the reporter gene. We made the dexamethasone-ABA bait for the screening of an ABA receptor. This bait inhibited the growth of rice, and was permeable to yeast cells. We assayed more than 10^6 of Arabidopsis cDNA clones, and observed their phenotypes to check the expression of reporter genes. In this meeting, we report experimental results obtained from the analysis of a positive clone.
植物化学調節学会の論文
2000-11-02