Abstract: Caffeic acid-O-methyltransferase（COMT） is a kind of enzyme that catalyzes oxygen atom on hydroxyl group of phenylpropanoid compounds. Previous studies had shown that F-box gene Glyma08G11030 encoding protein of soybean drought stress resistance gene may interact with COMT protein Glyma04G227700. In order to predict and analyze the function of soybean COMT gene Glyma04G227700 and the interaction between them. In this study, we cloned Glyma04G227700 gene and analyzed it by bioinformatics method. And then we analyzed the expression of Glyma04G227700 gene in different soybean tissues by real-time fluorescence quantitative detection method, and verified the protein interaction by yeast two-hybrid system. The results showed that the total length of Glyma04G227700 gene was 1 095 bp, encoding 366 amino acids, and it was closely related to the wild soybean（RZC17879.1 Glycine Soja）. Glyma04G227700 was expressed in the root, stem, leaf and cotyledon of soybean, and the expression level was higher in root and stem, and the lowest in cotyledon. There was no interaction between Glyma08G11030 and Glyma04G227700 in yeast double hybrid results.