CAJ | 학술논문

为提高黄酮代谢途径中的关键酶活性,增加苦荞黄酮类化合物的合成量,根据已知植物的F3H基因同源序列,采用RT-PCR和RACE结合的方法,克隆了苦荞黄酮代谢途径中关键酶基因F3H,命名为FtF3H(GenBank登录号:FJ456858).序列分析结果表明,FtF3H基因长1 369 bp, 编码367个氨基酸, 该氨基酸序列及其cDNA序列与葡萄、牵牛F3H等的同源性约80%.二级结构预测表明,随机卷曲是FtF3H蛋白最大量的结构元件,而α-螺旋和β-片层则散布于整个蛋白质中;三维结构建模预测FtF3H蛋白具有铁离子结合位点及2-O-酮戊二酸结合位点等F3H蛋白典型结构.
위제고황동대사도경중적관건매활성,증가고교황동류화합물적합성량,근거이지식물적F3H기인동원서렬,채용RT-PCR화RACE결합적방법,극륭료고교황동대사도경중관건매기인F3H,명명위FtF3H(GenBank등록호:FJ456858).서렬분석결과표명,FtF3H기인장1 369 bp, 편마367개안기산, 해안기산서렬급기cDNA서렬여포도、견우F3H등적동원성약80%.이급결구예측표명,수궤권곡시FtF3H단백최대량적결구원건,이α-라선화β-편층칙산포우정개단백질중;삼유결구건모예측FtF3H단백구유철리자결합위점급2-O-동무이산결합위점등F3H단백전형결구.
For the purpose of enhancing the key enzymes' activity to increase flavonoids synthesis in the biosynthetic pathway of flavonoids,one of the most important key enzymes flavanone 3-hydroxylase (F3H) gene was cloned from Fagopyrum tataricum.Based on cDNA sequence conserved domain of F3H,a pair of primers were designed and used to amplify a fragment of FtF3H gene (GenBank accession:FJ456858) using RT-PCR and RACE technique.A 1 369 bp full-length cDNA sequence was obtained.Analysis of F3H cDNA indicated that it encoded a peptide containing 367 amino acids.The sequence identity to other species was all about 80%.The predicted secondary structure demonstrated that random coil were the most important structural conformation.However,alpha helix and beta pleated sheet distributed in the whole protein.The predicted tertiary structure demonstrated that FtF3H has 2-oxoglutarate iron combination sites.