苦荞类异黄酮还原酶基因ftirl的克隆及序列分析.pdfVIP

210 2012, Vol. 33, No. 11 食品科学 ※生物工程 (FtIRL) 赵海霞，李成磊，白悦辰，陈 惠，吴 琦 * ( 四川农业大学生命科学与理学院，四川 雅安 625014) 利用 RT-PCR 技术，从苦荞(Fagopyrum tatarium) 中克隆得到类异黄酮还原酶基因(IRL) 的开放阅读框序列 (ORF) ，命名为FtIRL 。序列分析表明：FtIRL 含一个长 942bp 的ORF ，编码313 个氨基酸，其氨基酸序列与金 荞异黄酮还原酶 FcIFR (GenBank 登录号ABV02071) 同源性最高，达到97% ；与其他植物IRL 同源性为44%～73% 。 生物信息学分析表明：FtIRL 推导的蛋白质含有典型的底物结合口袋和保守的NADPH 结合位点，符合短链脱氢酶 家族的结构特征。构建基于氨基酸的系统发育树表明，苦荞 FcIFR 虽然在氨基酸序列上与其他植物的异黄酮还原酶 (IFR)有较高的同源性，但其生物学功能可能更接近落叶松脂醇还原酶(PLR) 。 苦荞；类异黄酮还原酶；基因克隆；序列分析 Cloning and Sequence Analysis of the Isoflavone Reductase-like (FtIRL) Gene from Fagopyrum tatarium ZHAO Hai-xia ，LI Cheng-lei ，BAI Yue-chen ，CHEN Hui ，WU Qi* (College of Life and Basic Sciences, Sichuan Agricultural University, Ya ’an 625014, China) Abstract ：The cDNA sequence of isoflavone reductase-like gene IRL from Fagopyrum tataricum was amplified by RT-PCR. Sequencing showed that the ORF of IRL gene from F . tataricum (named as FtIRL) was 942bp, probably encoding a protein of , 313 amino acids. Further analysis indicated that the deduced FcIFR protein had homology values of 97% with other plants IRLs. FtIRL possessed two short-chain dehydrogenase/reductase conserved motifs: a substrate-binding pocket and a NADPH binding site. The phylogenetic tree constructed based on amino acid sequences indicated that, although the amino acid sequence of IRL gene from F. tataricum had high homology with that of isoflavone reductase (IFR) genes from other plants, its biological function might be more similar to that of pinoresinol lariciresinol reductase (PLR). Key words ：Fagopyrum tataricum ；isoflavone reductase-like