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转录组的组装与表征

assembly and characterization of the transcriptome.

作者信息

Tong Guang-Xiang, Xu Wei, Zhang Yong-Quan, Zhang Qing-Yu, Yin Jia-Sheng, Kuang You-Yi

机构信息

Heilongjiang River Fisheries Research Institute of Chinese Academy of Fishery Sciences Daoli District, Harbin Heilongjiang Province China.

出版信息

Ecol Evol. 2017 Dec 21;8(2):1271-1285. doi: 10.1002/ece3.3735. eCollection 2018 Jan.

Abstract

Taimen () is an important ecological and economic species that is classified as vulnerable by the IUCN Red List of Threatened Species; however, limited genomic information is available on this species. RNA-Seq is a useful tool for obtaining genetic information and developing genetic markers for nonmodel species in addition to its application in gene expression profiling. In this study, we performed a comprehensive RNA-Seq analysis of taimen. We obtained 157 M clean reads (14.7 Gb) and used them to assemble a high-quality transcriptome with a N50 size of 1,060 bp. In the assembly, 82% of the transcripts were annotated using several databases, and 14,666 of the transcripts contained a full open reading frame. The assembly covered 75% of the transcripts of Atlantic salmon and 57.3% of the protein-coding genes of rainbow trout. To learn about the genome evolution, we performed a systematic comparative analysis across 11 teleosts including eight salmonids and found 313 unique gene families in taimen. Using Atlantic salmon and rainbow trout transcriptomes as the background, we identified 250 positive selection transcripts. The pathway enrichment analysis revealed a unique characteristic of taimen: It possesses more immune-related genes than Atlantic salmon and rainbow trout; moreover, some genes have undergone strong positive selection. We also developed a pipeline for identifying microsatellite marker genotypes in samples and successfully identified 24 polymorphic microsatellite markers for taimen. These data and tools are useful for studying conservation genetics, phylogenetics, evolution among salmonids, and selective breeding for threatened taimen.

摘要

哲罗鲑()是一种重要的生态和经济鱼类,被世界自然保护联盟濒危物种红色名录列为易危物种;然而,关于该物种的基因组信息有限。RNA测序是获取遗传信息和开发非模式物种遗传标记的有用工具,此外还可用于基因表达谱分析。在本研究中,我们对哲罗鲑进行了全面的RNA测序分析。我们获得了1.57亿条clean reads(14.7Gb),并用于组装一个高质量的转录组,N50大小为1060bp。在组装过程中,82%的转录本使用多个数据库进行了注释,其中14666个转录本包含完整的开放阅读框。该组装覆盖了大西洋鲑75%的转录本和虹鳟57.3%的蛋白质编码基因。为了了解基因组进化,我们对包括8种鲑科鱼类在内的11种硬骨鱼进行了系统的比较分析,在哲罗鲑中发现了313个独特的基因家族。以大西洋鲑和虹鳟的转录组为背景,我们鉴定出250个正选择转录本。通路富集分析揭示了哲罗鲑的一个独特特征:它拥有比大西洋鲑和虹鳟更多的免疫相关基因;此外,一些基因经历了强烈的正选择。我们还开发了一个用于鉴定样本中微卫星标记基因型的流程,并成功鉴定出24个哲罗鲑多态微卫星标记。这些数据和工具对于研究保护遗传学、系统发育学、鲑科鱼类之间的进化以及濒危哲罗鲑的选择性育种具有重要意义。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/257e/5773338/60618239a67c/ECE3-8-1271-g001.jpg

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