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RNA-seq Profiles of Immune Related Genes in the Spleen of Necrotic Enteritis-afflicted Chicken Lines

机译:坏死性肠炎患鸡脾脏中免疫相关基因的RNA-seq图谱

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The study aimed to compare the necrotic enteritis (NE)-induced transcriptome differences between the spleens of Marek's disease resistant chicken line 6.3 and susceptible line 7.2 co-infected with Eimeria maxima/Clostridium perfringens using RNA-Seq. Total RNA from the spleens of two chicken lines were used to make libraries, generating 42,736,296 and 42,617,720 usable reads, which were assembled into groups of 29,897 and 29,833 mRNA genes, respectively. The transcriptome changes were investigated using the differentially expressed genes (DEGs) package, which indicated 3,255, 2,468 and 2,234 DEGs of line 6.3, line 7.2, and comparison between two lines, respectively (fold change >= 2, p0.01). The transcription levels of 14 genes identified were further examined using qRT-PCR. The results of qRT-PCR were consistent with the RNA-seq data. All of the DEGs were analysed using gene ontology terms, the Kyoto Encyclopedia of Genes and Genomes (KEGG) database and the DEGs in each term were found to be more highly expressed in line 6.3 than in line 7.2. RNA-seq analysis indicated 139 immune related genes, 44 CD molecular genes and 150 cytokines genes which were differentially expressed among chicken lines 6.3 and 7.2 (fold change >= 2, p0.01). Novel mRNA analysis indicated 15,518 novel genes, for which the expression was shown to be higher in line 6.3 than in line 7.2 including some immune-related targets. These findings will help to understand host-pathogen interaction in the spleen and elucidate the mechanism of host genetic control of NE, and provide basis for future studies that can lead to the development of marker-based selection of highly disease-resistant chickens.
机译:这项研究的目的是比较使用RNA测序技术将马立克氏病抗性鸡品系6.3和易感品系7.2的感染情况与坏死性肠炎(NE)引起的转录组差异,该鸡系与艾美球虫/产气荚膜梭菌共感染。使用来自两个鸡系脾脏的总RNA来制作文库,生成42,736,296和42,617,720个可用读段,分别将其组装为29,897个和29,833个mRNA基因的组。使用差异表达基因(DEGs)软件包研究了转录组的变化,该数据包分别指示第6.3行,第7.2行的3,255、2,468和2,234 DEG,以及两行之间的比较(倍数变化> = 2,p <0.01)。使用qRT-PCR进一步检查了鉴定出的14个基因的转录水平。 qRT-PCR的结果与RNA-seq数据一致。使用基因本体术语,《京都基因与基因组百科全书》(KEGG)数据库对所有DEG进行了分析,发现每个术语中的DEG在6.3行中比在7.2行中的表达更高。 RNA-seq分析显示139个免疫相关基因,44个CD分子基因和150个细胞因子基因在鸡品系6.3和7.2之间差异表达(倍数变化≥2,p <0.01)。新型mRNA分析表明,有15518个新基因,在6.3行中其表达高于在7.2行中的表达,包括一些免疫相关靶标。这些发现将有助于了解脾脏中的宿主-病原体相互作用,并阐明NE的宿主遗传控制机制,并为将来的研究提供基础,这些研究可能导致开发基于标记的高度抗病鸡。

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