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Identification and comparative analysis of the Pseudosciaena crocea microRNA transcriptome response to poly(I:C) infection using a deep sequencing approach

机译:使用深度测序方法鉴定和比较分析大隐假单胞菌对多聚(I:C)感染的大隐假单胞菌microRNA转录组反应

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摘要

Two sRNA libraries with or without poly(I:C) infection of large yellow croaker Pseudosciaena crocea were constructed and sequenced using the high-throughput Illumina/Solexa deep sequencing technology. The high-throughput sequencing pipeline yielded 163,79,272 and 217,07,070 raw reads corresponding to 132,27,594 and 206,86,409 clean reads for the normal and infected libraries, respectively. Bioinfromatic analysis identified 534 miRNAs, of which, 158 miRNAs were known in miRBase 20.0 and the remaining 376 were not found homology to any known metazoan miRNAs, suggesting a possible species-specificity. We analyzed the significance of differently expressed miRNAs between two libraries using pairwise comparison. There was significant differential expression of 112 miRNAs (p < 0.001) between two libraries. Thereinto, a number of known miRNAs were identified immune-related. Real-time quantitative PCR experiments (RT-qPCR) were preformed for 6 miRNAs of the two samples, and agreement was found between the sequencing and RT-qPCR data. To our knowledge, this is the first comprehensive study of miRNAs in P. crocea and of expression analysis of P. crocea miRNAs in response to poly(I:C) infection, and many miRNAs were differentially regulated under normal and infection conditions. These findings deepened our understanding of the role of miRNAs in the intricate host's immune system, and should be useful to develop new control strategies for host immune defense against various foreign infection in P. crocea
机译:使用高通量Illumina / Solexa深度测序技术,构建了两个带有或不带有大黄鱼(Poseudosciaena crocea)的poly(I:C)感染的sRNA文库,并进行了测序。高通量测序流水线分别产生163,79,272和217,07,070原始读取,分别对应于正常和受感染文库的132,27,594和206,86,409干净读取。生物信息学分析鉴定了534个miRNA,其中在miRBase 20.0中已知158个miRNA,而其余376个未与任何已知的后生动物miRNA同源,表明可能存在物种特异性。我们使用成对比较分析了两个文库之间差异表达的miRNA的重要性。两个文库之间有112个miRNA的显着差异表达(p <0.001)。其中,鉴定出许多已知的miRNA与免疫相关。对两个样品的6个miRNA进行了实时定量PCR实验(RT-qPCR),并且在测序和RT-qPCR数据之间发现了一致性。据我们所知,这是对番红花中的miRNA的首次全面研究,也是对番(I:C)感染做出反应的番红花miRNA的表达分析,在正常和感染条件下,许多miRNA受到差异调节。这些发现加深了我们对miRNA在复杂的宿主免疫系统中的作用的理解,并且对于开发针对宿主抗大红景天的各种外源性免疫防御的新控制策略应该是有用的。

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