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沙漠虹鳉(Melanotaenia splendida tatei)的从头转录组组装与注释,并与未来气候的候选基因进行比较。

De novo transcriptome assembly and annotation for the desert rainbowfish (Melanotaenia splendida tatei) with comparison with candidate genes for future climates.

作者信息

Gates Katie, Sandoval-Castillo Jonathan, Bernatchez Louis, Beheregaray Luciano B

机构信息

Molecular Ecology Lab, Flinders University, Adelaide 5001, South Australia, Australia.

Institut de Biologie Intégrative et des Systèmes, Université Laval, Québec, QC G1V 0A6, Canada.

出版信息

Mar Genomics. 2017 Oct;35:63-68. doi: 10.1016/j.margen.2017.05.008. Epub 2017 May 22.

Abstract

Transcriptomics via RNA-seq has rapidly emerged as a powerful tool for ecological and evolutionary studies, enabling genome-scale studies of adaptation via regulation of global gene expression. Here we present a de novo transcriptome for the desert rainbowfish (Melanotaenia splendida tatei) based on individuals sampled in the Lake Eyre Basin, Australia's arid zone river system. Recently developed methods in RNA-seq and bioinformatics were used for sequencing, assembling and annotating a high-quality liver transcriptome suitable for studies of ecology and adaptation in desert rainbowfish. Transcript annotation in UniprotKB using BLASTx assigned unique protein matches to ~47% of 116,092 Trinity genes, while BLASTp assigned unique protein matches to ~35% of 62,792 predicted protein-coding regions. A full Trinotate annotation report is provided for predicted genes and their corresponding transcripts. Annotations were compared with previously identified genes for transcriptional regulation and heritable plasticity in future climates in the subtropical rainbowfish (M. duboulayi), finding ~57% of these candidate genes present in the desert rainbowfish transcriptome. We discuss the utility of transcriptomics methods for ecological studies of adaptation, while emphasising a range of methodological considerations for dealing with transcriptome datasets. This newly assembled transcriptome is expected to help elucidate mechanisms for adaptation to high temperatures and a variable climate in desert aquatic fauna.

摘要

通过RNA测序进行的转录组学已迅速成为生态和进化研究的强大工具,能够通过全球基因表达调控进行基因组规模的适应性研究。在此,我们基于在澳大利亚干旱区河流系统艾尔湖盆地采集的样本,为沙漠虹鳉(Melanotaenia splendida tatei)构建了一个从头转录组。RNA测序和生物信息学中最近开发的方法被用于测序、组装和注释一个高质量的肝脏转录组,该转录组适用于沙漠虹鳉的生态学和适应性研究。使用BLASTx在UniprotKB中进行转录本注释,为116,092个Trinity基因中的约47%分配了独特的蛋白质匹配,而使用BLASTp为62,792个预测的蛋白质编码区域中的约35%分配了独特的蛋白质匹配。为预测基因及其相应转录本提供了完整的Trinotate注释报告。将注释与先前在亚热带虹鳉(M. duboulayi)中鉴定的用于未来气候下转录调控和遗传可塑性的基因进行比较,发现沙漠虹鳉转录组中存在约57%的这些候选基因。我们讨论了转录组学方法在适应性生态研究中的实用性,同时强调了处理转录组数据集的一系列方法学考虑因素。这个新组装的转录组有望有助于阐明沙漠水生动物适应高温和多变气候的机制。

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