Cloning and Analysis of a Chalone Synthase Gene from Fagopyrum tataricum  

苦荞中查尔酮合成酶全长基因的克隆及序列分析(英文)

作　　者：刘凯[1,2] 胡耀辉[1] 王冠[3] 于寒松[1] 

机构地区：[1]吉林农业大学食品科学与工程学院,吉林长春130118 [2]吉林省农业综合开发办公室,吉林长春130118 [3]吉林农业大学生命科学学院,吉林长春130118

出　　处：《Agricultural Science & Technology》2012年第4期708-710,726,共4页农业科学与技术（英文版）

基　　金：Supported by 948 Program,Ministry of Agriculture of China(2008-z27)~~

摘　　要：[Objective] This study aimed to clone a full-length CHS gene from buck- wheat. [Method] With total RNA extracted from buckwheat as the template, CHS cD- NA sequence was cloned from buckwheat by using RACE technology and CODEHOP primer design method, the full length gene was obtained by primers which were de- signed for amplification of full-length gene sequence with buckwheat DNA template. Clustalxl.81 and MEGA4 software were used for sequence analysis and construction of phylogenetic tree; NCBI Blastn and Biastp programs were applied for homology analysis of nucleic acid and protein. [Result] Bioinformatics analysis showed that the full length of this gene is 1 906 bp, containing a 463 bp intron sequence and a 1 188 bp coding region, encoding 395 amino acids. Blastn sequence alignment revealed that the CHS gene sequence obtained in this study shared 86% homology with the CHS gene of closely related species. [Conclusion] This study laid the foundation to clarify molecular basis of the synthesis of buckwheat bioflavanoids and explore an effective way to improve the content of buckwheat bioflavanoids.[目的]该研究旨在克隆苦荞中查尔酮合成酶全长基因。[方法]选用乌克兰伊琳娜苦荞为试验材料,以从叶片中提取的RNA为模板,应用RACE技术结合CODEHOP引物设计方法克隆苦荞中查尔酮合成酶cDNA序列,通过电子合并获得其全长。设计基因全长特异性引物,以DNA为模板进行PCR扩增出基因序列。应用Clustalxl.81和MEGA4软件进行序列分析和进化树的建立;核酸和蛋白质序列同源性分析应用NCBI的Blastn和Blastp完成。[结果]生物信息学分析表明,该基因全长1906bp,具有一个463bp的内含子序列,编码区长度为1188bp,编码395个氨基酸。Blastn序列比对发现该试验所获得的CHS基因序列与相近物种Rheum palmatum(登录号:DQ205352.1)的CHS基因同源性达86%。[结论]该研究为阐明苦荞生物类黄酮合成的分子基础,探索提高苦荞生物类黄酮含量的有效途径奠定基础。

关 键 词：Fagopyrum tataricum CHS RACE INTRON Cladogram 

分 类 号：S517[农业科学—作物学]