日本血吸虫童虫miRNA高通量测序分析

来源 :中华地方病学杂志 | 被引量 : 0次 | 上传用户:heatsink
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目的:高通量测序分析日本血吸虫童虫的微小RNA(microRNA,miRNA),鉴定已知miRNA的表达水平,预测分析miRNA靶基因及其生物学功能。方法:体外制备日本血吸虫童虫,提取童虫的总RNA构建文库进行Illumina高通量测序。采用DEGseq R语言包,结合perl脚本进行miRNA表达量的差异分析;分别利用miRanda、Blast软件和KEGG数据库对差异miRNA进行靶基因及其生物学功能预测。结果:构建文库中,日本血吸虫童虫表达的miRNAs与最新miRBase数据库比对结果显示,共有38 483条匹配序列,鉴定到已知miRNA 60个;其中,sja-miR-125b表达量最高,其次为sja-miR-61、sja-miR-71a、sja-miR-36-3p和sja-miR-10-5p,上述5种miRNA表达量占总miRNA的91%(3 263/3 585)。共预测到靶基因7 176个,基因功能集中在核苷酸转移酶活性、细胞氮复合代谢、分子功能、生物学过程、生物合成、等离子体膜及蛋白质成熟;功能富集分析显示,高表达的miRNA主要参与致病过程、生物进展及多条代谢途径通路。结论:日本血吸虫童虫显著表达的miRNA参与了血吸虫分化、生长发育和致病过程中的代谢途径调节,为研究血吸虫发育调控机制及新药物开发奠定了基础。“,”Objective:To identify the expression level of known microRNA (miRNA) by high-throughput sequencing and analysis of the miRNA of n Schistosoma japonicum schistosomula, and predict the miRNA target genes and their biological functions.n Methods:Schistosoma japonicum schistosomula were prepared n in vitro, and total RNA of schistosomula were extracted and analyzed to construct a library for performing high-throughput sequencing. The difference of miRNA expression was analyzed by using DEGseq R language package and perl script. Then the target genes and their biological functions of differential miRNA were predicted by miRanda software, Blast software, and KEGG database respectively.n Results:There were 38 483 matching sequences in the miRNA expressed of n Schistosoma japonicum schistomula in the constructed library compared with the latest miRBase database, and 60 miRNA were identified, sja-miR-125b was the miRNA with the highest expression, followed by sja-miR-61, sja-miR-71a, sja-miR-36-3p and sja-miR-10-5p, which accounted for 91% (3 263/3 585) of the total miRNA expression. MiRanda software predicted a total of 7 176 target genes, gene functions concentrated on nucleotide transferase activity, cellular nitrogen complex metabolism, molecular function, biological processes, biosynthesis, plasma membrane and protein maturation. The functional enrichment analysis showed that the highly expressed miRNA were mainly involved in pathogenic process, biological progress and multiple metabolic regulation pathways.n Conclusions:The miRNA expressed significantly of n Schistosoma japonicum schistomula mainly involved in the regulation of metabolic pathways during the differentiation, growth and pathogenesis of n Schistosoma japonicum. To lay a foundation for the study of the regulatory mechanism of n Schistosoma japonicum development and the development of new drugs.n
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