苦荞中查尔酮合成酶全长基因的克隆及序列分析(英文)

[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。

Cloning and Analysis of a Chalone Synthase Gene from Fagopyrum tataricum

[Objective] This study aimed to clone a full-length CHS gene from buckwheat.[Method] With total RNA extracted from buckwheat as the template,CHS cDNA sequence was cloned from buckwheat by using RACE technology and CODEHOP primer design method,the full length gene was obtained by primers which were designed 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 Blastp 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.