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翡翠灰蛾,扁叶蜂(鞘翅目,多食亚目)中 mariner 样元件的分子特征。

Molecular characterization of mariner-like elements in emerald ash borer, Agrilus planipennis (Coleoptera, Polyphaga).

机构信息

Department of Entomology, The Ohio State University/Ohio Agricultural and Research Development Center, Wooster, Ohio 44691, USA.

出版信息

Arch Insect Biochem Physiol. 2010 Aug;74(4):205-16. doi: 10.1002/arch.20357.

Abstract

Emerald ash borer (EAB, Agrilus planipennis), an exotic invasive pest, has killed millions of ash trees (Fraxinus spp.) in North America and continues to threaten the very survival of the entire Fraxinus genus. Despite its high-impact status, to date very little knowledge exists for this devastating insect pest at the molecular level. Mariner-like elements (MLEs) are transposable elements, which are ubiquitous in occurrence in insects and other invertebrates. Because of their low specificity and broad host range, they can be used for epitope-tagging, gene mapping, and in vitro mutagenesis. The majority of the known MLEs are inactive due to in-frame shifts and stop codons within the open reading frame (ORF). We report on the cloning and characterization of two MLEs in A. planipennis genome (Apmar1 and Apmar2). Southern analysis indicated a very high copy number for Apmar1 and a moderate copy number for Apmar2. Phylogenetic analysis revealed that both elements belong to the irritans subfamily. Based on the high copy number for Apmar1, the full-length sequence was obtained using degenerate primers designed to the inverted terminal repeat (ITR) sequences of irritans MLEs. The recovered nucleotide sequence for Apmar1 consisted of 1,292 bases with perfect ITRs, and an ORF of 1,050 bases encoding a putative transposase of 349 amino acids. The deduced amino acid sequence of Apmar1 contained the conserved regions of mariner transposases including WVPHEL and YSPDLAP, and the D,D(34)D motif. Both Apmar1 and Apmar2 could represent useful genetic tools and provide insights on EAB adaptation.

摘要

外来入侵害虫翡翠灰螟(EAB,Agrilus planipennis)已在北美洲杀死了数百万株白蜡树(Fraxinus spp.),并继续威胁整个白蜡属的生存。尽管其具有高度的破坏性,但迄今为止,在分子水平上对这种破坏性的昆虫害虫的了解非常有限。Mariner 样元件(MLEs)是转座元件,在昆虫和其他无脊椎动物中普遍存在。由于其特异性低和宿主范围广,它们可用于表位标记、基因作图和体外诱变。已知的大多数 MLEs 由于在开放阅读框(ORF)内发生移框和终止密码子而处于非活性状态。我们报告了在 A. planipennis 基因组中克隆和表征两个 MLEs(Apmar1 和 Apmar2)的情况。Southern 分析表明 Apmar1 的拷贝数非常高,而 Apmar2 的拷贝数适中。系统发育分析表明这两个元件都属于 irritans 亚科。基于 Apmar1 的高拷贝数,使用针对 irritans MLEs 的反向末端重复(ITR)序列设计的简并引物获得了全长序列。Apmar1 的回收核苷酸序列由 1,292 个碱基组成,具有完美的 ITR,ORF 为 1,050 个碱基,编码一个假定的 349 个氨基酸的转座酶。Apmar1 的推导氨基酸序列包含 mariner 转座酶的保守区域,包括 WVPHEL 和 YSPDLAP,以及 D,D(34)D 基序。Apmar1 和 Apmar2 都可能成为有用的遗传工具,并为翡翠灰螟的适应提供见解。

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