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暹罗鱼虱从头转录组分析:甲壳动物 Toll 下游信号分子特征的首次分子研究。

De novo whole transcriptome analysis of the fish louse, Argulus siamensis: first molecular insights into characterization of Toll downstream signalling molecules of crustaceans.

机构信息

Central Institute of Freshwater Aquaculture, Kausalyaganga, Bhubaneswar 751002, India.

出版信息

Exp Parasitol. 2013 Nov;135(3):629-41. doi: 10.1016/j.exppara.2013.09.018. Epub 2013 Sep 30.

DOI:10.1016/j.exppara.2013.09.018
PMID:24090566
Abstract

Argulus siamensis is a major ectoparasitic pathogen of freshwater fish capable of causing substantial economic loss. None of the available control measures have been able to address the problem of argulosis resourcefully. To combat this pathogen effectively, it is necessary to have a comprehensive understanding of its life processes with information on various genes involved. The transcriptome studies can generate introductory information about genes participating in physiological processes of the parasite which could be targeted for their control. In this study, the transcriptome sequencing of A. siamensis was performed on Illumina HiSeq 2000 platform which generated 75,126,957 high quality reads. A total of 46,352 transcript contigs were assembled with average length of 1211bp and N50 length of 2302bp. In total, 19,290 CDS including 184 novel CDS and 59,019 open reading frames (ORFs) were identified from the assembled contigs. Gene ontology and Kyoto Encylopedia of Genes and Genomes pathway analysis were performed to classify contigs into their functional categories and regulation pathways. Additionally, 1171 simple sequence repeats were identified from the assembled contigs. Further, twelve contigs with high similarity with downstream molecules of the mammalian toll like receptor (TLR) pathway were validated by their inductive expressions in response to lipopolysaccharide (LPS) of Gram negative bacteria, Escherichia coli and Gram positive bacteria, Staphylococcus aureus. The transcriptome of an ectoparasite A. siamensis was sequenced, assembled, annotated, and the downstream signalling molecules of Toll pathway characterized. The transcriptome data generated will facilitate studies on functional genomics that will subsequently be applied for vaccine development and other control strategies against the parasite.

摘要

暹罗鱼怪是一种严重的淡水鱼类外寄生虫病原体,能够造成巨大的经济损失。目前尚无任何可用的控制措施能够巧妙地解决鱼怪病问题。为了有效地对抗这种病原体,我们需要全面了解其生命过程,包括涉及的各种基因信息。转录组研究可以为寄生虫生理过程中参与的基因提供初步信息,这些基因可能成为控制的靶点。在这项研究中,我们在 Illumina HiSeq 2000 平台上对 A. siamensis 的转录组进行了测序,生成了 75126957 条高质量reads。共组装出 46352 条转录本contigs,平均长度为 1211bp,N50 长度为 2302bp。总共鉴定出 19290 个 CDS,包括 184 个新的 CDS 和 59019 个开放阅读框(ORFs)。从组装的 contigs 中进行了基因本体和京都基因与基因组百科全书通路分析,将 contigs 分类为其功能类别和调控通路。此外,还从组装的 contigs 中鉴定出 1171 个简单序列重复。进一步地,通过对革兰氏阴性菌、大肠杆菌和革兰氏阳性菌、金黄色葡萄球菌的脂多糖(LPS)诱导表达,验证了 12 条与哺乳动物 toll 样受体(TLR)途径下游分子具有高相似性的 contigs。对暹罗鱼怪这种外寄生虫的转录组进行了测序、组装、注释,并对 Toll 途径的下游信号分子进行了表征。生成的转录组数据将促进功能基因组学的研究,随后将应用于疫苗开发和其他针对寄生虫的控制策略。

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