二化螟Ty3/gypsy反转座子的克隆与序列分析

doi:10.3969/j.issn.1001-7216.2014.03.011

中国水稻科学 ›› 2014, Vol. 28 ›› Issue (3): 314-321.DOI: 10.3969/j.issn.1001-7216.2014.03.011

二化螟Ty3/gypsy反转座子的克隆与序列分析

李晓欢^{1, 2, ＃},罗光华^{2, ＃},张志春²,刘宝生²,方继朝^{1, 2,*}

¹南京农业大学植物保护学院，南京 210095; ²江苏省农业科学院植物保护研究所，南京 210014；

收稿日期:2014-02-14 修回日期:2014-03-19 出版日期:2014-05-10 发布日期:2014-05-10
通讯作者: 方继朝1, 2,*
基金资助:
国家水稻产业技术体系项目（Cars00125); 国家自然科学基金资助项目（31201505）；江苏省农业科技自主创新资金资助项目（CX(13)5025)。

Cloning and Characterization of Ty3/gypsy Retrotransposon in Chilo suppressalis (Lepidoptera: Pyralidae)

LI Xiaohuan^{1, 2, ＃}, LUO Guanghua^{2, ＃}, ZHANG Zhichun², LIU Baosheng², FANG Jichao^{1, 2,*}

¹ College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China; ²Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China;

Received:2014-02-14 Revised:2014-03-19 Online:2014-05-10 Published:2014-05-10
Contact: FANG Jichao1, 2,*

摘要/Abstract

摘要： 转座子是宿主基因组的重要组成部分。Ty3/gypsy反转座子是广泛存在于生物体内的一类反转座子。通过反向PCR（inverse PCR）和基因组步移方法成功地从二化螟体内获得一个具有完整结构的Ty3/gypsy反转座子拷贝，命名为CsuTy3（GenBank登录号：KJ191261）。该反转座子拷贝全长4 934 bp，在基因组上插入的靶位点（target sit duplications，TSDs）是“AACGT”，两端的长末端重复序列（long terminal repeat，LTR）不完全相同，5′端LTR长161 bp，3′端LTR长168 bp，两者的一致度为93.5%；在靠近3′端LTR处有一段长15 bp的嘌呤尾（polypurine tract，PPT）序列。经ORF Finder分析，CsuTy3中包含三个独立的ORF，从左至右第一个ORF编码类似病毒的结构蛋白，称为GAG蛋白；第二个ORF编码天冬氨酰蛋白酶（aspartic protease，AP）蛋白；第三个ORF编码一个多聚蛋白，包含反转录酶（reverse transcriptase，RT)、RNA酶H（RNase H，RH）以及整合酶（integrase，IN）。Southern杂交显示二化螟不同地理种群中均有较多的CsuTy3拷贝。在特定基因座位上，对CsuTy3拷贝插入与否的检测结果显示，二化螟不同地理种群中在该位点上均含有CsuTy3拷贝的插入，但在德阳和江津种群中有些个体在该位点上没有检测到CsuTy3拷贝的插入。

关键词: 二化螟, 反转座子, Ty3/gypsy, 克隆, 序列分析

Abstract: Transposable elements constitute a substantial fraction of host genomes. Ty3/gypsy retrotransposon, one group of LTR retrotransposons, is widespread in different species. By inversePCR and genome walking, a novel member of Ty3/gypsy retrotransposons was cloned from Chilo suppressalis. This new member of Ty3/gypsy retrotransposons was named CsuTy3 (GenBank accession No. KJ191261). The sequence is 4 934 bp in length and integrated into the “AACGT” target site duplications (TSDs) of the genome. There is a pair of noncompletely identical long terminal repeats (LTRs) in the CsuTy3 retrotransposon. The 5′LTR is 161 bp in length and the 3′LTR is 168 bp, sharing 935% similarities. A polypurine tract (PPT), 15 bp in length, is adjacent to the 3′LTR. There are three independent open reading frames (ORFs) in CsuTy3. The first ORF encodes a protein which is related to viral structural protein, termed GAG. The second encodes aspartic protease (AP). The third encodes a polyprotein including a reverse transcriptase (RT) which produces a cDNA copy of the transposon′s RNA, an RNase H (RH) which splits the DNARNA hybrid and an integrase (IN) which inserts the cDNA into the host′s genome. The southern hybridization indicated that there were many CsuTy3 copies in different C. suppressalis populations. Flanking PCR results showed that the CsuTy3 copy was inserted at the same locus in different populations. At this locus, all the individuals have the CsuTy3 copy insertion except some individuals from Deyang and Jiangjin populations. The CsuTy3 insertion ratio varied with field populations.

Key words: Chilo suppressalis, retrotransposon, Ty3/gypsy, cloning, sequence analysis

中图分类号:

李晓欢1, 2, ＃,罗光华2, ＃,张志春2,刘宝生2,方继朝1, 2,*. 二化螟Ty3/gypsy反转座子的克隆与序列分析[J]. 中国水稻科学, 2014, 28(3): 314-321.

LI Xiaohuan1, 2, ＃, LUO Guanghua2, ＃, ZHANG Zhichun2, LIU Baosheng2, FANG Jichao1, 2,*. Cloning and Characterization of Ty3/gypsy Retrotransposon in Chilo suppressalis (Lepidoptera: Pyralidae)[J]. Chinese Journal of Rice Science, 2014, 28(3): 314-321.

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二化螟Ty3/gypsy反转座子的克隆与序列分析

Cloning and Characterization of Ty3/gypsy Retrotransposon in Chilo suppressalis (Lepidoptera: Pyralidae)

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