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园艺学报 ›› 2014, Vol. 41 ›› Issue (1): 157-164.

• 研究报告 • 上一篇    下一篇

牡丹LINE 类反转录转座子RT 序列的克隆及分析

宋程威1,郭大龙2,3,张 曦1,郭丽丽1,3,侯小改1,3,*   

  1. 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;3 河南省高校牡丹工程技术
    中心,河南洛阳 471003
  • 出版日期:2014-01-25 发布日期:2014-01-25
  • 基金资助:

    国家自然科学基金项目(31070620);洛阳市科技攻关项目(1102061A);河南省高校科技创新人才支持计划项目
    (13HASTIT004);河南省重点科技攻关项目(132102110029)

Cloning and Analysis of Reverse Transcriptase of LINE-retrotransposons of Tree Peony(Paeonia

SONG Cheng-wei1,GUO Da-long2,3,ZHANG Xi1,GUO Li-li1,3,and HOU Xiao-gai1,3,*   

  1. 1College of Agriculture,Henan University of Science & Technology,Luoyang,Henan 471003,China;2College of
    Forestry,Henan University of Science & Technology,Luoyang,Henan 471003,China;3Engineering Technology Research
    Center of Tree Peony of Henan Province University,Luoyang,Henan 471003,China
  • Online:2014-01-25 Published:2014-01-25

摘要: 利用LINE 简并引物从中原牡丹品种‘洛阳红’中扩增出580 bp 左右的目的条带,对其回
收、克隆、测序及相关生物信息学软件分析后获得32 条LINE 类牡丹反转录转座子RT(反转录酶)序列。
这些序列长度为547 ~ 593 bp,主要表现为点突变,经Clustal W 比对其同源性为25.7% ~ 94.2%。将其核
苷酸序列经聚类后可分为4 个家族,其中家族Ⅰ和 Ⅲ 分别占总序列数的50%和28%。将32 条RT 序列翻
译成氨基酸序列,在第18 氨基酸序列处有1 个非常保守的的甘氨酸(Gly),在138 氨基酸序列处有1 个
半保守的赖氨酸(Lys)位点。32 条序列均发生较多的终止密码子突变和移码突变。与其他物种的系统进
化树分析,表明牡丹LINE 类反转录转座子RT 序列既有一定的保守性,同时也与其他物种间存在一定的
同源性。

关键词: 牡丹, 非LTR 类反转录转座子, 反转录酶

Abstract: A fragment of 580 bp was amplified by PCR from the genomic DNA of tree peony
(Paeonia suffruticosa Andrews.‘Luoyanghong’)using the degenerate oligoncleotide primers. The
amplicons were recovered,purified and cloned. Positive clones were identified and sequenced. Finally,
32 different sequences of reverse transcriptase from tree peony‘Luoyanghong’LINE retrotransposons
were obtained. The length of the nucleotide sequences varied from 547 to 593 bp. The homology ranged
from 25.7% to 94.2% by Clustal W and the variations were characterized by point mutation. Four clusters
were identified with high heterogeneity through phylogenic analysis of their nucleotide sequences. Family
Ⅰand Ⅲ,respectively accounted for 50% and 28% of the total number of clones. There is a very conservative point in the 18th amino acid sequence of glycine(Gly)in the translated sequence,at the same
time,a conservative point in 138th amino acid sequence of lysine(Lys). More termination codon mutation
and frameshift mutations were presented in 32 sequences. A phylogenic tree was constructed based on the
amino acid sequences from other species , indicating that the RT sequences from Peony LINE
retrotransposons have certain homology with other species.

Key words: tree peony, non-LTR retrotranspson, reverse transcriptase

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