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通过靶向转录组焦磷酸测序技术开发太平洋鲱基因组资源。

Development of genomic resources for Pacific Herring through targeted transcriptome pyrosequencing.

机构信息

School of Aquatic and Fishery Sciences, University of Washington, Seattle, Washington, United States of America.

出版信息

PLoS One. 2012;7(2):e30908. doi: 10.1371/journal.pone.0030908. Epub 2012 Feb 27.

Abstract

Pacific herring (Clupea pallasii) support commercially and culturally important fisheries but have experienced significant additional pressure from a variety of anthropogenic and environmental sources. In order to provide genomic resources to facilitate organismal and population level research, high-throughput pyrosequencing (Roche 454) was carried out on transcriptome libraries from liver and testes samples taken in Prince William Sound, the Bering Sea, and the Gulf of Alaska. Over 40,000 contigs were identified with an average length of 728 bp. We describe an annotated transcriptome as well as a workflow for single nucleotide polymorphism (SNP) discovery and validation. A subset of 96 candidate SNPs chosen from 10,933 potential SNPs, were tested using a combination of Sanger sequencing and high-resolution melt-curve analysis. Five SNPs supported between-ocean-basin differentiation, while one SNP associated with immune function provided high differentiation between Prince William Sound and Kodiak Island within the Gulf of Alaska. These genomic resources provide a basis for environmental physiology studies and opportunities for marker development and subsequent population structure analysis.

摘要

太平洋鲱(Clupea pallasii)支持商业和文化上重要的渔业,但也受到各种人为和环境因素的额外压力。为了提供基因组资源,以促进机体和种群水平的研究,对来自威廉王子湾、白令海和阿拉斯加湾的肝和睾丸样本的转录组文库进行了高通量焦磷酸测序(罗氏 454)。鉴定出超过 40000 个序列,平均长度为 728bp。我们描述了一个注释的转录组以及单核苷酸多态性(SNP)发现和验证的工作流程。从 10933 个潜在的 SNP 中选择了 96 个候选 SNP 的子集,使用 Sanger 测序和高分辨率熔解曲线分析相结合的方法进行了测试。5 个 SNP 支持了海洋盆地之间的分化,而与免疫功能相关的 1 个 SNP 在阿拉斯加湾的威廉王子湾和科迪亚克岛之间提供了高度的分化。这些基因组资源为环境生理学研究提供了基础,并为标记物的开发和随后的种群结构分析提供了机会。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a13b/3288011/2d2758d80ef8/pone.0030908.g001.jpg

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