萨能奶山羊胎儿成纤维细胞miRNA文库的构建及生物信息学分析

本试验旨在丰富山羊miRNA文库，并探讨miRNAs在胎儿成纤维细胞生长调控中的作用。试验以萨能奶山羊胎儿成纤维细胞为研究对象，运用Illumina高通量测序和生物信息学技术分析山羊胎儿成纤维细胞miRNAs表达。成功构建了萨能奶山羊胎儿成纤维细胞miRNAs的cDNA文库，获得16 395 039 total reads，去除冗余数据后获得16 150 181条clean reads，其中unique sRNAs 205 857条。生物信息学分析获得候选新miRNAs 247条，候选miRNAs靶基因8 401个，靶基因位点数10 832个；同时分析发现候选miRNAs首位碱基对U和A具有偏向性。GO注释表明，69.6%的基因与细胞和细胞组分相关，超过54.1%的基因参与了细胞器相关活动，多达65.0%的基因与细胞及细胞过程相关联。KEGG通路分析显示，约10.5%的基因与代谢通路有关，是占比最多的一条通路，数据显示miRNAs对胎儿成纤维细胞具有重要调控作用。试验结果为胎儿成纤维细胞的生长调控及奶山羊乳腺生物反应器中供核体细胞miRNAs深入研究提供参考。

Construction of miRNA Library and Bioinformatics Analysis About Fetal Fibroblasts of Saanen Dairy Goat

The purpose of this experiment was to enrich miRNA library of goat and explore the role of miRNAs in the regulation of cell growth.The expression of Saanen dairy goat fetal fibroblast miRNAs was analyzed by Illumina high-throughput sequencing and bioinformatics technology.Total miRNAs cDNA library from fibroblasts of Saanen dairy goat was successfully constructed,16 395 039 total reads and 16 150 181 clean reads had been obtained and among them were 205 857 unique sRNAs.From those clean reads,247 novel miRNAs were identified and 8 401 target genes were predicted with 10 832 target sites.Meanwhile,the first base pairs of candidate miRNAs were biased for U and A.GO analysis revealed that more than 69.6% of the genes were related to cells and cellular components,more than 54.1% of the genes were involved in organelle related activity,and approximately 65.0% of the genes were related with cells and cellular processes.KEGG pathway analysis showed that approximately 10.5% of the genes were related to a metabolic pathway,and that it was the pathway that accounts for the most.In conclusion,the data showed that miRNAs played an important regulatory role in fetal fibroblasts.The results provided a basis for further research on the growth regulation of fetal fibroblasts and miRNAs of donor cells in the breast bioreactor of dairy goats.