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RNA-Seq 分析揭示了与凡纳滨对虾对 Taura 综合征病毒(TSV)抗性相关的基因。

RNA-Seq analysis reveals genes associated with resistance to Taura syndrome virus (TSV) in the Pacific white shrimp Litopenaeus vannamei.

机构信息

Center of Excellence for Molecular Biology and Genomics of Shrimp, Department of Biochemistry, Faculty of Science, Chulalongkorn University, 254 Phayathai Road, Bangkok 10330, Thailand.

出版信息

Dev Comp Immunol. 2013 Dec;41(4):523-33. doi: 10.1016/j.dci.2013.07.020. Epub 2013 Aug 3.

Abstract

Outbreak of Taura syndrome virus (TSV) is one of the major pathogens of the Pacific white shrimp Litopenaeus vannamei. Although selective breeding for improvement of TSV resistance in L. vannamei has been successfully developed and has led to a great benefit to the shrimp farming industry worldwide. The molecular mechanisms underlying the viral resistance in shrimp remain largely unknown. In the present study, we conducted the first transcriptomic profiling of host responses in hemolymph and hemocytes in order to identify the differentially expressed genes associated with resistance to TSV in L. vannamei. High-throughput RNA-Seq was employed, obtaining 193.6 and 171.2 million high-quality Illumina reads from TSV-resistant and susceptible L. vannamei lines respectively. A total of 61,937 contigs were generated with an average length of 546.26 bp. BLASTX-based gene annotation (E-value < 10(-5)) allowed the identification of 12,398 unique proteins against the NCBI non-redundant NR database. In addition, comparison of digital gene expression between resistant and susceptible strains revealed 1374 significantly differentially expressed contigs (representing 697 unigenes). Gene pathway analysis of the differentially expressed gene set highlighted several putative genes involved in the immune response activity including (1) pathogen/antigen recognition including immune regulator, adhesive protein and signal transducer; (2) coagulation; (3) proPO pathway cascade; (4) antioxidation; and (5) protease. The expression patterns of 22 differentially expressed genes involving immune response were validated by quantitative real-time RT-PCR (average correlation coefficients 0.94, p-value < 0.001). Our results provide valuable information on gene functions associated with resistance to TSV in L. vannamei.

摘要

对虾 Taura 综合征病毒(TSV)的爆发是斑节对虾(Litopenaeus vannamei)的主要病原体之一。尽管对斑节对虾进行了针对 TSV 抗性的选择育种,并且已经给全球的对虾养殖业带来了巨大的利益,但虾类对病毒抗性的分子机制在很大程度上仍不清楚。在本研究中,我们进行了对虾血淋巴和血细胞中宿主反应的首次转录组谱分析,以鉴定与斑节对虾对 TSV 抗性相关的差异表达基因。我们采用高通量 RNA-Seq 技术,分别从 TSV 抗性和易感斑节对虾品系中获得了 19360 万和 17120 万条高质量的 Illumina reads。总共生成了 61937 个 contigs,平均长度为 546.26bp。基于 BLASTX 的基因注释(E 值 < 10(-5)) 允许对 12398 个独特的蛋白质进行鉴定,这些蛋白质针对 NCBI 非冗余 NR 数据库。此外,对抗性和易感品系之间的数字基因表达进行比较,发现了 1374 个显著差异表达的 contigs(代表 697 个 unigenes)。差异表达基因集的基因途径分析突出了几个可能参与免疫反应活性的基因,包括(1)病原体/抗原识别,包括免疫调节剂、黏附蛋白和信号转导物;(2)凝血;(3)proPO 途径级联;(4)抗氧化;和(5)蛋白酶。通过定量实时 RT-PCR 验证了 22 个涉及免疫反应的差异表达基因的表达模式(平均相关系数 0.94,p 值 < 0.001)。我们的研究结果为斑节对虾对 TSV 抗性相关基因功能提供了有价值的信息。

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