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南极软毛海豹(Arctocephalus gazella)皮肤转录组中的基因发现。

Gene discovery in the Antarctic fur seal (Arctocephalus gazella) skin transcriptome.

机构信息

Department of Animal Behaviour, University of Bielefeld, Postfach 100131, 33501 Bielefeld, Germany.

出版信息

Mol Ecol Resour. 2011 Jul;11(4):703-10. doi: 10.1111/j.1755-0998.2011.02999.x. Epub 2011 Mar 16.

Abstract

Next-generation sequencing provides a powerful new approach for developing functional genomic tools for nonmodel species, helping to narrow the gap between studies of model organisms and those of natural populations. Consequently, massively parallel 454 sequencing was used to characterize a normalized cDNA library derived from skin biopsy samples of twelve Antarctic fur seal (Arctocephalus gazella) individuals. Over 412 Mb of sequence data were generated, comprising 1.4 million reads of average length 286 bp. De novo assembly using Newbler 2.3 yielded 156 contigs plus 22 869 isotigs, which in turn clustered into 18,576 isogroups. Almost half of the assembled transcript sequences showed significant similarity to the nr database, revealing a functionally diverse array of genes. Moreover, 97.9% of these mapped to the dog (Canis lupis familiaris) genome, with a strong positive relationship between the number of sequences locating to a given chromosome and the length of that chromosome in the dog indicating a broad genomic distribution. Average depth of coverage was also almost 20-fold, sufficient to detect several thousand putative microsatellite loci and single nucleotide polymorphisms. This study constitutes an important step towards developing genomic resources with which to address consequential questions in pinniped ecology and evolution. It also supports an earlier but smaller study showing that skin tissue can be a rich source of expressed genes, with important implications for studying the genomics not only of marine mammals, but also more generally of species that cannot be destructively sampled.

摘要

下一代测序为非模式物种开发功能基因组工具提供了一种强大的新方法,有助于缩小模型生物研究与自然种群研究之间的差距。因此,我们使用大规模平行 454 测序技术对来自 12 只南极软毛海豹(Arctocephalus gazella)个体皮肤活检样本的归一化 cDNA 文库进行了测序。共生成了超过 4.12 亿个序列数据,包含平均长度为 286bp 的 140 万个读数。使用 Newbler 2.3 进行从头组装得到 156 个 contigs 和 22869 个 isotigs,它们进一步聚类为 18576 个 isogroups。组装的转录序列中几乎有一半与 nr 数据库具有显著相似性,揭示了一系列功能多样的基因。此外,这些基因中有 97.9%映射到犬(Canis lupis familiaris)基因组,在给定染色体上定位的序列数量与该染色体在犬中的长度之间存在很强的正相关关系,这表明了广泛的基因组分布。平均覆盖深度也几乎是 20 倍,足以检测到数千个假定的微卫星位点和单核苷酸多态性。本研究是朝着开发基因组资源以解决鳍足类生态学和进化中的重要问题迈出的重要一步。它还支持了之前的一项但规模较小的研究,表明皮肤组织是表达基因的丰富来源,这对于研究不仅是海洋哺乳动物,而且更普遍地是那些不能进行破坏性采样的物种的基因组学具有重要意义。

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