摘要
【目的】鉴定感染鸡白痢沙门氏菌后雏鸡脾脏差异表达的miRNA,为阐明miRNA在鸡白痢沙门氏菌感染中的调控机制提供理论基础。【方法】采集感染和未感染鸡白痢沙门氏菌的SPF雏鸡脾脏,提取脾脏RNA,构建6个miRNA文库;利用Illumina Hiseq 2500测序技术和生物信息学分析筛选差异表达miRNA,通过TargetScan和miRanda算法预测差异表达miRNA的靶基因,并进行GO功能富集和KEGG通路富集分析,最后利用RT-qPCR方法验证测序结果。【结果】鉴定到29个差异表达miRNAs,其中15个显著上调、14个显著下调。GO分析表明:脂多糖介导的信号通路正调控以及NIK/NF-κB介导的信号通路负调控等免疫相关生物学过程显著富集;KEGG分析显示:差异表达的miRNA主要参与溶酶体和内吞作用等免疫相关通路。高通量测序结果与RT-qPCR结果一致。【结论】成功鉴定到鸡白痢沙门氏菌感染雏鸡脾脏miRNA表达谱,获得29个鸡白痢沙门氏菌感染潜在相关miRNAs。
[Purpose]To identify the differentially expressed miRNA in spleen of chicken infected with Salmonella Pullorum,providing a theoretical basis for clarifying the regulatory mechanism of miRNA in Salmonella Pullorum infection.[Methods]Spleens from SPF chicks infected and uninfected with Salmonella Pullorum were collected,spleen RNA was extracted,and six miRNA libraries were constructed.The differentially expressed miRNA were screened by Illumina Hiseq 2500 highthroughput sequencing technology and bioinformatics technology,the target genes were predicted by TargetScan and miRanda,and the predicted target genes were analyzed using GO functional enrichment and KEGG pathway enrichment.Finally,RT-qPCR was used to verify the sequencing results.[Results]A total of 29 differentially expressed miRNAs were obtained,including 15 significantly up-regulated and 14 significantly down-regulated miRNAs.Immune-related biological processes such as positive regulation of lipopolysaccharide-mediated signaling pathways and negative regulation of NIK/NF-κB-mediated signaling pathways were significantly enriched in GO analysis.KEGG pathway enrichment showed that differentially expressed miRNA were mainly involved in immune-related pathways such as lysosome and endocytosis.In addition,the results of high-throughput sequencing were consistent with those of RT-qPCR.[Conclusion]The miRNA expression profile of chicks spleen infected with Salmonella Pullorum is successfully identified,and 29 potential miRNAs related to Salmonella Pullorum infection are obtained.
作者
白潜
陈远红
陈曦
江康峰
叶玮琪
向斌
杨亮宇
杨静
BAI Qian;CHEN Yuanhong;CHEN Xi;JIANG Kangfeng;YE Weiqi;XIANG Bin;YANG Liangyu;YANG Jing(College of Veterinary Medicine,Yunnan Agricultural University,Kunming 650201,China)
出处
《云南农业大学学报（自然科学版）》
CAS
CSCD
北大核心
2023年第2期220-227,共8页
Journal of Yunnan Agricultural University:Natural Science
基金
云南省万人计划-产业技术领军人才(YNWR-CYJS-2019-020)
国家蛋鸡产业技术体系(CARS-40-S25)。
