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花生基因组中活性微型反向重复转座元件的特征。

Characterization of active miniature inverted-repeat transposable elements in the peanut genome.

机构信息

Department of Plant Genome Research, Kazusa DNA Research Institute, 2-6-7 Kazusa-Kamatari, Kisarazu, Chiba 292-0818, Japan.

出版信息

Theor Appl Genet. 2012 May;124(8):1429-38. doi: 10.1007/s00122-012-1798-6.

Abstract

Miniature inverted-repeat transposable elements (MITEs), some of which are known as active nonautonomous DNA transposons, are found in the genomes of plants and animals. In peanut (Arachis hypogaea), Ah-MITE1 has been identified in a gene for fatty-acid desaturase, and possessed excision activity. However, the AhMITE1 distribution and frequency of excision have not been determined for the peanut genome. In order to characterize AhMITE1s, their genomic diversity and transposition ability was investigated. Southern blot analysis indicated high AhMITE1 copy number in the genomes of A. hypogaea, A. magna and A. monticola, but not in A. duranensis. A total of 504 AhMITE1s were identified from the MITE-enriched genomic libraries of A. hypogaea. The representative AhMITE1s exhibited a mean length of 205.5 bp and a GC content of 30.1%, with AT-rich, 9 bp target site duplications and 25 bp terminal inverted repeats. PCR analyses were performed using primer pairs designed against both flanking sequences of each AhMITE1. These analyses detected polymorphisms at 169 out of 411 insertional loci in the four peanut lines. In subsequent analyses of 60 gamma-irradiated mutant lines, four Ah-MITE1 excisions showed footprint mutations at the 109 loci tested. This study characterizes AhMITE1s in peanut and discusses their use as DNA markers and mutagens for the genetics, genomics and breeding of peanut and its relatives.

摘要

微型反向重复转座元件(MITEs),其中一些被称为活跃的非自主 DNA 转座子,存在于植物和动物的基因组中。在花生(Arachis hypogaea)中,已经在脂肪酸去饱和酶基因中鉴定出 Ah-MITE1,并具有切除活性。然而,尚未确定花生基因组中 AhMITE1 的分布和切除频率。为了表征 AhMITE1,研究了它们的基因组多样性和转座能力。Southern blot 分析表明,A. hypogaea、A. magna 和 A. monticola 基因组中的 AhMITE1 拷贝数较高,但 A. duranensis 基因组中没有。从 A. hypogaea 的 MITE 富集基因组文库中鉴定出总共 504 个 AhMITE1。代表性的 AhMITE1 平均长度为 205.5 bp,GC 含量为 30.1%,具有富含 AT 的 9 bp 靶位点重复和 25 bp 末端反向重复。使用针对每个 AhMITE1 侧翼序列设计的引物对进行 PCR 分析。这些分析在四个花生品系的 411 个插入位点中的 169 个检测到多态性。在随后对 60 个γ射线诱变突变体系的分析中,四个 Ah-MITE1 切除在测试的 109 个位点显示出足迹突变。本研究对花生中的 AhMITE1 进行了表征,并讨论了它们作为 DNA 标记物和诱变剂在花生及其亲缘关系的遗传学、基因组学和育种中的应用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a8f/3336055/ae083dc5bf9e/122_2012_1798_Fig1_HTML.jpg

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