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评估黑森瘿蚊基因组中 Desmar 样元件插入位点的结构变异和分子作图。

Assessment of structural variation and molecular mapping of insertion sites of Desmar-like elements in the Hessian fly genome.

机构信息

Eck Institute for Global Health, Department of Biological Sciences, University of Notre Dame, Notre Dame, IN 46556, USA.

出版信息

Insect Mol Biol. 2010 Dec;19(6):707-15. doi: 10.1111/j.1365-2583.2010.01028.x.

Abstract

The Hessian fly (Mayetiola destructor) is an agriculturally important pest of wheat. A mariner element (Desmar1) has been previously identified in the Hessian fly genome. Using Desmar1 as a probe, we isolated individual copies of Desmar-like elements from the Hessian fly genome cloned in bacterial artificial chromosomes (BACs) and studied their structural variability and flanking DNA sequences. The partial Desmar-like copies are relatively more abundant (∼64%) than full length copies (∼36%) in the Hessian fly genome. Most of the full length copies are consistently flanked by an EcoRI restriction site that occurs 32 bp from one end and 66 bp from the other end of the mariner. Using an amplified fragment length polymorphism-PCR (AFLP-PCR) based method, we identified segregating polymorphisms associated with Desmar elements in a F₂ mapping population. We were able to use the segregation data to localize the chromosomal position of three Desmar elements by linkage analysis. As paternal chromosomes are eliminated in the Hessian fly during early embryogenesis, two-thirds of the AFLPs were expected to be polymorphic in the mapping population and this was observed for AFLPs anchored to full length Desmar copies but not to the partial copies. Thus, our data indicate that dead and partial Desmar-like copies are probably associated with less polymorphic regions and may represent mariner graveyards in the Hessian fly genome.

摘要

麦长管蚜(Mayetiola destructor)是一种对小麦具有重要农业意义的害虫。先前在麦长管蚜基因组中已鉴定出海员元件(Desmar1)。我们使用 Desmar1 作为探针,从麦长管蚜基因组克隆的细菌人工染色体(BAC)中分离出单个的 Desmar 样元件,并研究了它们的结构变异性和侧翼 DNA 序列。部分 Desmar 样拷贝在麦长管蚜基因组中比全长拷贝更丰富(约 64%)。大多数全长拷贝始终被 EcoRI 限制位点侧翼,该位点距海员的一端 32 个碱基,距另一端 66 个碱基。使用基于扩增片段长度多态性-PCR(AFLP-PCR)的方法,我们在 F₂ 作图群体中鉴定出与 Desmar 元件相关的分离多态性。我们能够通过连锁分析利用分离数据定位三个 Desmar 元件的染色体位置。由于在早期胚胎发生过程中,麦长管蚜中的父本染色体被消除,因此三分之二的 AFLP 在作图群体中预计是多态的,这在锚定到全长 Desmar 拷贝的 AFLP 中观察到,但在部分拷贝中没有观察到。因此,我们的数据表明,死的和部分的 Desmar 样拷贝可能与多态性较低的区域相关,并且可能代表麦长管蚜基因组中的海员墓地。

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