大豆Glyma04G227700生物信息学分析及与Glyma08G11030互作研究

Bioinformatics Analysis of Soybean Glyma04G227700 and Studies of Interaction with Glyma08G11030

咖啡酸-O-甲基转移酶(caffeic acid-O-methyltransferase, COMT)是一种催化苯丙素类化合物羟基上氧原子的甲基化酶。前期研究表明,大豆抵御干旱胁迫的F-box基因Glyma08G11030编码蛋白可能与COMT蛋白Glyma04G227700发生互作,为了预测分析大豆COMT基因Glyma04G227700的功能及二者的互作关系,本研究克隆Glyma04G227700基因,并对其进行生物信息学分析,通过实时荧光定量检测方法分析其在大豆不同组织中的表达情况,并利用酵母双杂交系统验证蛋白互作情况。结果显示:Glyma04G227700基因全长1 095 bp,编码366个氨基酸,与野大豆(RZC17879.1 Glycine soja)亲缘关系较近。Glyma04G227700在大豆的根、茎、叶、子叶中都有表达,且在根和茎中表达量较高,在子叶中表达量最低。酵母双杂结果表明Glyma08G11030与Glyma04G227700蛋白不存在互作。

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.