Screening And Identification Of Cellular MicroRNAs Involved In The Infection Of Goatpox Virus And Its Application In Early Detection

micro RNAs(mi RNAs) are a subclass of small RNAs with approximately 22 nt in length and repress the stability or translation of the m RNAs of the target genes through binding of the 3’UTR, 5’UTR or protein-coding region of the transcripts. Accelerateing evidences revealed that mi RNAs are involved in diverse biological processes, including cellular proliferation, differentiation, apoptosis and viral infections. However, few studies have concentrated on the links between mi RNA and Goatpox virus(GPV) infection. It is necessary for us to decipher the roles of mi RNAs in GPV infection.We have screened the differentially expressed mi RNAs using Illumina/Solexa high throughput sequencing plantform and performed the prediction of target genes of mir-365-3p and mir-365-5p. According to the Gene onthology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) analysis of the target genes, we studied the roles of mir-365-5p and mir-365-3p in celluar processes. Finally, we discussed the capability of mir-365-3p and mir-365-5p that served as the potential biomarkers for early detection of GPV infection. The main results are as follows:(1) The primary Lamb Testicle(LT) cell was made. The titer of GPV GH strain was determined and the TCID50 value was 10-4.67/100μL.(2) The gene expression variation of Dicer and Argonaute-2(AGO-2) was determined with q PCR. The results revealed the significant upregulation of Dicer m RNA.(3) 91 differentially expressed mi RNAs were identified by mi RNA sequencing plantform. 48 of these mi RNAs are upregulated, and the remaining mi RNAs are downregulated.(4) Identification of the expression type of mir-365-3p and mir-365-5p from 0h to 48 h. The results revealed that the mir-365-5p was downregulated during the period. mir-365-3p was upregulated during the first 10 h, however, mir-365-3p was downregulated from 10 h to 48 h.(5) The putative target genes of mir-365-3p and mir-365-5p were characterized with Target Scan database. According to the GO analysis, we revealed that the molecular functions of the target genes of mir-365-3p were enriched in protein binding, DNA binding and regulation of the activities of transcription factors. The biological processes the target genes of mir-365-3p were enriched in regulation of transcription and cellular basal metabolism. The molecular functions of the target genes of mir-365-5p were mainly enriched in protein binding and the biological processes of the target genes were enriched in protein metabolism. The KEGG analysis uncovered that the target genes of mir-365-3p were significantly enriched in regulation of cytoskeleton, chemokine pathways, Wnt signaling pathway and mitogen-activated protein kinase(MAPK) pathway. And the target genes of mir-365-5p were enriched in ubiquitin-mediated proteolytic pathway and Erb B signaling pathway.(6) Identification the expression variation of mir-365-3p and mir-365-5p of diverse tissues of sheep after 3 days of inoculation with GPV. The results revealed that mir-365-3p was downregulated in liver, lung and rumen and had the potential to serve as biomarkers for early dectection for GPV infection.