欧李ChUFGT基因的克隆、生物信息学分析及亚细胞定位

Cloning,Bioinformatics Analysis and Subcellular Localization of ChUFGT Gene from Cerasus humilis

为从分子水平上揭示欧李中3-O-类黄酮葡萄糖基转移酶(UFGT)基因的空间表达情况,以欧李叶片为试材,根据欧李的转录组序列设计特异性引物,通过PCR获得ChUFGT基因序列,并进行生物信息学分析及亚细胞定位分析。结果表明,ChUFGT基因DNA开放阅读框为1425 bp(不含终止子),相对分子质量为51.63 kD,编码475个氨基酸,其对应的蛋白质是带正电荷的稳定疏水性蛋白。与近缘物种的蛋白质序列比对分析显示,欧李、李和樱桃李的UFGT氨基酸序列相似度达到97.22%和96.62%,整个系统发育树存在两个大分支,其中ChUFGT与同科植物的亲缘关系最近,其次是与白梨、西洋梨、荔枝等其他科植物相对较近,与葡萄、刺葡萄、石榴等植物的亲缘关系最远。ChUFGT的亚细胞定位结果分析显示,在洋葱内表皮细胞的细胞核和细胞质中都表达该基因。本研究结果为进一步阐明ChUFGT调控欧李细胞花色素苷的合成机制提供了理论基础。

In order to reveal the spatial expression of the 3-o-flavonoid glucosyltransferase(UFGT)gene in Cerasus humilis at the molecular level,specific primers were designed according to the transcriptome sequence of Cerasus humilis,and the gene sequence was obtained by PCR,and the bioinformatics and subcellular localization expression analysis were carried out.The results showed that the open reading frame of ChUFGT gene was 1425 bp(Without terminator),its relative molecular weight was 51.63 kD,encoding 475 amino acids.Its corresponding protein was a stable hydrophobic protein with positive charge.Comparison with the protein sequences of related species showed that the similarity of UFGT amino acid sequences of Cerasus humilis,Prunus salicina and Prunus cerasifera reached 97.22%and 96.62%.There are two major branches of the phylogenetic tree,among which ChUFGT is the most closely related to plants of the same family,followed by white pear,western pear,litchi and other other families,and is the most furthest to grape,thorn grape,pomegranate and other plants.The results of subcellular localization analysis showed that the ChUFGT was expressed in both the nucleus and cytoplasm of onion inner epidermal cells.The results of this study was to further clarify ChUFGT regulation Cerasus humilis cells anthocyan nucleoside synthesis mechanism provides a theoretical basis.