牡丹Ty3-gypsy类反转录转座子反转录酶序列的克隆及分析  

Cloning and Analysis of Reverse Trancriptase of Ty3-gypsy-like Retrotransposons in Tree Peony(Paeonia)

作　　者：侯小改[1] 郭大龙[2] 黄燕梅[2] 张曦[1] 

机构地区：[1]河南科技大学农学院,河南洛阳471003 [2]河南科技大学林学院,河南洛阳471003

出　　处：《园艺学报》2013年第1期98-106,共9页Acta Horticulturae Sinica

基　　金：国家自然科学基金项目(31070620);河南省高校科技创新人才支持计划项目(2010HASTTT002);河南省高等学校青年骨干教师资助计划项目(2010GGJS-072)

摘　　要：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。Using degenerate oligoncleotide primers corresponding to conserved domains of the Ty3-gypsy-like retrotransposon reverse transcriptase，a fragment of 430 bp was amplified by PCR from the genomic DNA of tree peony（Paeonia suffruticosa Andrews‘Luoyang Hong’）and Paeonia qiui. The amplicons were recovered and cloned into pMD-18T vector after purification，positive clones were selected and identified by colony PCR，then sequenced and analyzed. Thirteen different sequences of reverse transcriptase from tree peony‘Luoyang Hong’and Paeonia qiui were obtained and six clusters were identified with high heterogeneity through phylogenic analysis after alignment analyses of their nucleotide sequences. These sequences showed high heterogeneity，mainly characterized by deletion mutations. The length of the nucleotide sequences varied from 412 to 446 bp，and homology ranged from 71.5% to 94.8%. When translated into amino acids，twelve sequences presented stop codon mutation，and three sequences presented frameshift mutation. A phylogenic tree was constructed based on the amino acid sequences from other species，indicating that horizontal transmission of retrotransposon has occurred among the plants in the past.

关 键 词：牡丹 Ty3-gypsy 类反转录转座子 反转录酶 异质性 

分 类 号：S685.11[农业科学—观赏园艺]