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MicroRNA (miRNA) are a recently discovered class of non-coding regulatory small molecule RNA plays an important regulatory role in the infection process. The purpose of this study is to find the pig vascular endothelial cells (SUVEC) in the microRNA, screened swine fever virus infection miRNA. (1) Application Illumina Solexa sequencing instrument in pig endothelial cells 18,721,836 35 nt small RNA sequences, a total of 13,612,934 of high quality by removing low quality and the linker sequence small RNA sequences (clean reads), and then through the SOAP software (Short oligonucleotide alignment program) these sRNA position to the genome, analysis of its genome distribution and expression of the case, results in 8,547,658 sequences (accounting for the total number of 62.79%) than on a pig genome. Through NCBI GenBank and the Rfam two databases, and repeat the sequence of intron and exon child compared remove the possible rRNA scRNA tRNA, snRNA, snoRNA from mRNA degradation fragment of sRNAs results obtained 5,932,143 a miRNA, 43.58% of the total sRNA. By bioinformatic analysis, we identified 154 miRNA genes encoding 116 mature conserved miRNAs and 66 miRNA * discovered four new pig-specific miRNAs. (2) using gene chip technology to confirm the discovery of miRNA expression, and filter out the pig vascular endothelial cells were seeded CSFV differentially expressed miRNAs, eight miRNAs were up-regulated, the expression levels of four miRNAs down. (3) Applied Biology Software RNA22 prediction found combination of miRNA let-7 family CSFV 3'UTR, and dual luciferase reporter gene vector containing CSFV3 'non-coding region fragment validation, the results show that the let-7a, let- 7c and let-7i combined with CSFV 3'UTR. Further confirm the regulatory role of the let-7 of classical swine fever virus, respectively, to the let-7 family, let-7a, let-7c and let-7i mimetics and inhibitors swine umbilical vein endothelial cells transfected into, and then inoculated with CSFV heterogeneous nuclear protein A1 (Nuclear ribonucleoproteins, hnRNP A1), let-7a and let-7c mimics to inhibit the proliferation of CSFV; cells after vaccination CSFV CSFV swine endothelial cells express increased expression of let-7a downgrades. The results of this study revealed the role of miRNA in vascular endothelial cells in the pig CSFV infection, molecular pathogenic mechanism of CSFV provide valuable scientific information.
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