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以五年生人参根组织为试验材料,利用RT-PCR方法克隆人参DS基因序列;通过生物信息学技术对其核苷酸序列和氨基酸序列进行比对;运用实时荧光定量PCR技术检测DS基因在人参根、茎、叶、果各部位的表达量。结果表明:获得人参DS基因c DNA全长,其核苷酸序列长为2.385 kb,含有1个开放阅读框,编码769个氨基酸多肽。生物信息学分析显示DS编码蛋白质包含1个跨膜区域,具有3个保守结构域。人参DS编码蛋白质与西洋参的DS(AGK62449)和三七的DS(AGI19258)具有99%的序列相似性。实时荧光定量PCR显示,DS基因在人参根、茎、叶、果各部位均有表达,在叶中表达量最高,其次是在果中,在根和茎中表达量相近。
The five-year-old ginseng root tissue was used as experimental material to clone the DS gene sequence of ginseng by RT-PCR. The nucleotide and amino acid sequences of the ginseng were cloned by bioinformatics technology. The real-time fluorescent quantitative PCR Root, stem, leaf, fruit of all parts of the expression. The results showed that the full length of cDNA of ginseng DS gene was obtained. Its nucleotide sequence was 2.385 kb in length and contained an open reading frame encoding a polypeptide of 769 amino acids. Bioinformatics analysis revealed that the DS-encoded protein contains 1 transmembrane region with 3 conserved domains. Ginseng DS-coded protein has 99% sequence similarity to DS (AGK62449) of Panax quinquefolium and DS (AGI19258) of Panax notoginseng. Real-time quantitative PCR showed that the DS gene was expressed in all parts of ginseng root, stem, leaf and fruit, the highest expression in leaf, the second was in fruit, and the same in roots and stems.