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转录组学研究巨型淡水虾(罗氏沼虾):从头组装、注释和标记发现。

Transcriptomics of a giant freshwater prawn (Macrobrachium rosenbergii): de novo assembly, annotation and marker discovery.

机构信息

Biogeosciences, Queensland University of Technology, Brisbane, Queensland, Australia.

出版信息

PLoS One. 2011;6(12):e27938. doi: 10.1371/journal.pone.0027938. Epub 2011 Dec 8.

Abstract

BACKGROUND

Giant freshwater prawn (Macrobrachium rosenbergii or GFP), is the most economically important freshwater crustacean species. However, as little is known about its genome, 454 pyrosequencing of cDNA was undertaken to characterise its transcriptome and identify genes important for growth.

METHODOLOGY AND PRINCIPAL FINDINGS

A collection of 787,731 sequence reads (244.37 Mb) obtained from 454 pyrosequencing analysis of cDNA prepared from muscle, ovary and testis tissues taken from 18 adult prawns was assembled into 123,534 expressed sequence tags (ESTs). Of these, 46% of the 8,411 contigs and 19% of 115,123 singletons possessed high similarity to sequences in the GenBank non-redundant database, with most significant (E value < 1e(-5)) contig (80%) and singleton (84%) matches occurring with crustacean and insect sequences. KEGG analysis of the contig open reading frames identified putative members of several biological pathways potentially important for growth. The top InterProScan domains detected included RNA recognition motifs, serine/threonine-protein kinase-like domains, actin-like families, and zinc finger domains. Transcripts derived from genes such as actin, myosin heavy and light chain, tropomyosin and troponin with fundamental roles in muscle development and construction were abundant. Amongst the contigs, 834 single nucleotide polymorphisms, 1198 indels and 658 simple sequence repeats motifs were also identified.

CONCLUSIONS

The M. rosenbergii transcriptome data reported here should provide an invaluable resource for improving our understanding of this species' genome structure and biology. The data will also instruct future functional studies to manipulate or select for genes influencing growth that should find practical applications in aquaculture breeding programs.

摘要

背景

巨型淡水虾(罗氏沼虾或 GFP)是最重要的经济淡水甲壳类动物物种。然而,由于对其基因组知之甚少,因此进行了 cDNA 的 454 焦磷酸测序,以描述其转录组并鉴定对生长重要的基因。

方法和主要发现

从 18 只成年对虾的肌肉、卵巢和精巢组织中制备的 cDNA 进行 454 焦磷酸测序分析获得了 787,731 条序列读段(244.37 Mb),将这些序列读段组装成 123,534 条表达序列标签(EST)。其中,8411 条拼接体中有 46%的拼接体和 115,123 条单序列的 19%与 GenBank 非冗余数据库中的序列具有高度相似性,最显著(E 值<1e(-5))的拼接体(80%)和单序列(84%)与甲壳类和昆虫序列匹配。拼接体开放阅读框的 KEGG 分析鉴定出了几个可能对生长重要的生物学途径的推定成员。检测到的顶级 InterProScan 域包括 RNA 识别基序、丝氨酸/苏氨酸蛋白激酶样结构域、肌动蛋白样家族和锌指结构域。在肌肉发育和构建中起重要作用的肌动蛋白、肌球蛋白重链和轻链、原肌球蛋白和肌钙蛋白等基因的转录本含量丰富。在拼接体中,还鉴定出了 834 个单核苷酸多态性、1198 个插入缺失和 658 个简单序列重复基序。

结论

本研究报道的 M. rosenbergii 转录组数据应该为我们理解该物种的基因组结构和生物学提供宝贵的资源。该数据还将指导未来的功能研究,以操纵或选择影响生长的基因,这些基因在水产养殖育种计划中应有实际应用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/17aa/3234237/adf9a3bc6a98/pone.0027938.g001.jpg

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