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

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.