苦荞中查尔酮合成酶全长基因的克隆及其序列分析  

Molecular Cloning and Analysis of a Chalone Synthase Gene from Fagopyrum tataricum

作　　者：胡耀辉[1] 王冠[2] 刘凯[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,长春130118 [2]吉林农业大学生命科学学院,长春130118 [3]吉林农业大学食品科学与工程学院和吉林省农业综合开发办公室,长春130118

出　　处：《生物技术通报》2011年第7期65-68,72,共5页Biotechnology Bulletin

基　　金：国家农业部"948"项目(2008-z27)

摘　　要：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引物设计方法成功克隆出苦荞中查尔酮合成酶cDNA序列后,将其序列电子合并其全长后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。通过生物信息学分析表明,该基因全长为1 906 bp,具有一个463 bp的内含子序列,编码区长度为1 188 bp,编码395个氨基酸。Blastn序列比对发现本试验所获得的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%。将得到的序列命名为RaCHS并提交GenBank,登录号为HQ434624。应用Clustalxl.81和MEGA4软件构建系统进化树,并进行了同源性分析。Buckwheat from shanxi as experiment material,total RNA was extracted from the leaves.Then the cDNA sequence of chalcone synthase was cloned by RACE combined with CODEHOP primer design method.The full-length cDNA CHS gene was amplified using DNA as template by PCR.Bioinformatics analysis show that the full-length of CHS gene sequence of Fagopyrum tataricum is 1 906 bp,including an intron which is 463 bp,and the open reading frame is 1 188 bp encoding 395 amino acids.Phylogenic analysis by the Blastn on NCBI showed that the identity to other Fagopyrum is 86%.The gene is named as RaCHS（GenBank accession number is HQ434624）.Phylogenetic tree constructed by the software of Clustal xl.81 and MEGA4.

关 键 词：苦荞 查尔酮合成酶 RACE 内含子 进化树 

分 类 号：S517[农业科学—作物学] Q943.2[生物学—植物学]