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Genome Evolution and Meiotic Maps by Massively Parallel DNA Sequencing: Spotted Gar, an Outgroup for the Teleost Genome Duplication

机译:基因组进化和减数分裂地图通过大规模平行的DNA测序:发现Gar,Teleost基因组复制的外群。

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摘要

Genomic resources for hundreds of species of evolutionary, agricultural, economic, and medical importance are unavailable due to the expense of well-assembled genome sequences and difficulties with multigenerational studies. Teleost fish provide many models for human disease but possess anciently duplicated genomes that sometimes obfuscate connectivity. Genomic information representing a fish lineage that diverged before the teleost genome duplication (TGD) would provide an outgroup for exploring the mechanisms of evolution after whole-genome duplication. We exploited massively parallel DNA sequencing to develop meiotic maps with thrift and speed by genotyping F1 offspring of a single female and a single male spotted gar (Lepisosteus oculatus) collected directly from nature utilizing only polymorphisms existing in these two wild individuals. Using Stacks, software that automates the calling of genotypes from polymorphisms assayed by Illumina sequencing, we constructed a map containing 8406 markers. RNA-seq on two map-cross larvae provided a reference transcriptome that identified nearly 1000 mapped protein-coding markers and allowed genome-wide analysis of conserved synteny. Results showed that the gar lineage diverged from teleosts before the TGD and its genome is organized more similarly to that of humans than teleosts. Thus, spotted gar provides a critical link between medical models in teleost fish, to which gar is biologically similar, and humans, to which gar is genomically similar. Application of our F1 dense mapping strategy to species with no prior genome information promises to facilitate comparative genomics and provide a scaffold for ordering the numerous contigs arising from next generation genome sequencing.
机译:由于组装良好的基因组序列的费用和多代研究的困难,无法获得数百种具有进化,农业,经济和医学重要性的基因组资源。硬骨鱼类提供了许多人类疾病的模型,但是拥有古老复制的基因组,有时会混淆连接性。代表在硬骨鱼基因组复制(TGD)之前发散的鱼类谱系的基因组信息将为探索全基因组复制后的进化机制提供一个整体。我们利用大规模平行的DNA测序技术,通过对仅从这两个野生个体中存在的多态性直接从自然界中收集的单头雌性和单头雄性斑gar(Lepisosteus oculatus)的F1后代进行基因分型,以节俭和速度发展出减数分裂图。使用Stacks(可通过Illumina测序分析从多态性中自动调用基因型的软件),我们构建了包含8406个标记的图谱。两个跨图幼虫上的RNA-seq提供了一个参考转录组,可识别近1000个定位的蛋白质编码标记,并允许对全基因组进行保守性分析。结果显示,在TGD及其基因组的组织方式与人类相比,骨之前,the氏谱系与硬骨动物背道而驰。因此,斑点雀gar提供了在硬骨鱼类(生物学上与gar类似)和人类(雀类在基因上相似)之间的医学模型之间的关键链接。将我们的F1密集作图策略应用于没有事先基因组信息的物种,有望促进比较基因组学的发展,并为订购下一代基因组测序产生的大量重叠群提供了一个支架。

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