基于转录组测序技术进行静原鸡差异miRNA与mRNA的关联分析

Correlation Analysis of Differential miRNA and mRNA in Jingyuan Chicken Based on Transcriptome Sequencing

为探究静原鸡肌肉组织肌苷酸沉积过程的分子调控机制,本研究以静原鸡胸肌和腿肌组织作为试验材料,通过转录组测序技术,利用生物信息学分析方法筛选出差异表达miRNA及其靶基因。分析结果表明,静原鸡胸肌和腿肌组织的比较组合中有39个差异表达miRNAs (19个显著上调, 20个显著下调)。通过miRNA-mRNA互作网络分析可知,一个miRNA可以靶向多个m RNA参与调控多个靶基因的表达。GO富集分析表明,候选靶基因的功能主要富集于三个分支的单一生物过程、细胞内和结构分子活性等。KEGG通路注释表明,候选靶基因显著富集到碳代谢、蛋白酶体通路和氨基酸的生物合成等通路中。选取4个差异表达miRNA的与肌苷酸合成和代谢相关的候选靶基因(POLR2C, GMPR, IMPDH2和APRT)进行实时荧光定量PCR验证,结果表明,定量结果与转录组测序结果趋势一致。该研究结果为阐明地方鸡种肌苷酸特异性沉积过程中miRNA介导的靶基因与肉品风味的关系及其表达分析提供理论依据。

To explore the molecular regulatory mechanism of the inosine acid deposition process in muscle tissue of Jingyuan chicken, chest muscle and leg muscle tissues of Jingyuan chicken were used as experimental materials in this study. The differentially expressed miRNA and its target genes were screened by transcriptome sequencing and bioinformatics analysis. The results showed that there were 39 different expressed miRNAs(19 significantly up-regulated, 20 significantly down-regulated) in the chest and leg muscles of Jingyuan chicken. Through the analysis of miRNA-mRNA interaction network, it can be seen that one miRNA can target multiple m RNA to participate in the regulation of multiple target gene expression. GO enrichment analysis showed that the functions of candidate target genes were mainly concentrated in the single biological process, intracellular and structural molecular activities of three branches. KEGG pathway annotation showed that candidate target genes were significantly enriched in carbon metabolism, proteasome pathway and amino acid biosynthesis. Four candidate target genes(POLR2C, GMPR, IMPDH2 and APRT) related to inosine synthesis and metabolism that differentially ex-pressed miRNA were selected for real-time fluorescence quantitative PCR validation. The results showed that the trend of quantitative PCR results was consistent with that of transcriptome sequencing. The results of this study provided a theoretical basis for elucidating the relationship between miRNA-mediated target genes and meat flavor and their expression analysis in the process of inosine specific deposition of local chicken species.