牡丹LINE类反转录转座子RT序列的克隆及分析  

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

作　　者：宋程威[1] 郭大龙[2,3] 张曦[1] 郭丽丽[1,3] 侯小改[1,3] 

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 [3]河南省高校牡丹工程技术中心,河南洛阳471003

出　　处：《园艺学报》2014年第1期157-164,共8页Acta Horticulturae Sinica

基　　金：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)

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。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 I and III, respectively accounted for 50% and 28% of the total number of clones. There is a veryconservative 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.

关 键 词：牡丹 非LTR类反转录转座子 反转录酶 

分 类 号：S685.11[农业科学—观赏园艺]