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根据 Ty3-gypsy 反转录转座子反转录酶的保守序列设计简并引物,从中原牡丹(Paeonia suffruticosa Andrews)品种‘洛阳红’和野生种卵叶牡丹(Paeonia qiui Y. L. Pei et D. Y. Hong)中扩增出430 bp 左右的目标片段。目的条带经回收、克隆、测序及相关生物信息学软件进行序列分析后,获得了13 条来自牡丹的 Ty3-gypsy 反转录转座子反转录酶序列。这些核苷酸序列具有较高的异质性,主要表现为缺失突变,序列长度变化范围为 412 ～ 446 bp,同源性范围为 71.5% ～ 94.8%。翻译成氨基酸后,有 12 条序列出现 1 ～ 9 个不同程度的终止密码子突变,3 条序列出现移框突变。其核苷酸序列经过系统聚类后可分为 6 个家族。将其氨基酸序列与已登录的不同物种 Ty3-gypsy 反转录转座子反转录酶的氨基酸序列进行聚类分析,结果表明与其他植物具有较高的同源性,表明它们间可能存在着 Ty3-gypsy 反转录转座子的横向传递。
The degenerate primers were designed according to the conserved sequence of Ty3-gypsy retrotransposon. Paeonia suffruticosa Andrews and Paeonia qiui YL Pei et DY Hong, In the amplification of 430 bp target fragment. Thirteen bands of Ty3-gypsy retrotransposon reverse transcriptase from Peony were obtained by sequence analysis after recovery, cloning, sequencing and related bioinformatics software. These nucleotide sequences have high heterogeneity, mainly showing deletion mutations, ranging in length from 412 bp to 446 bp in length and ranging from 71.5% to 94.8% in homology. After translating into amino acids, there are 12 stop codons of varying degrees in 12 sequences and three of them have a frame-shifting mutation. Its nucleotide sequence after system clustering can be divided into six families. Cluster analysis of the amino acid sequence and the amino acid sequence of Ty3-gypsy transposon reverse transcriptase of different species registered showed that there was a high homology with other plants, indicating that there may exist between them Lateral transmission of Ty3-gypsy retrotransposons.