Primary Selection MicroRNA Acting On CSFV In Swine Umbilical Vein Endothelial Cells

In recent years, microRNAs (miRNAs) which were discovered in eucaryote were single-stranded small noncoding RNAs that play important regulated roles in life activities. The microRNAs generally act on the non-coding region of mRNA, which repress the mRNA translation by imperfect complementary pairs and result in gene silence. Recent studies displayed that the microRNAs have important regulatory function in virus infection,so they maybe have key effect on CSFV infection through the interaction with 3′and5′-UTR of CSFV. Some miRNAs potentially targeted to CSFV genome were predicted by bioinformatics analysis. To sdudy the function of microRNA in CSFV infection, CSFV 3′-and 5′-untranslation region were cloned and then were inserted into psiCHECHTM-2 plasmid. The activity of Firefly and Renilla luciferases was detected by Luminometer to select the microRNA intracted with CSFV, which lay a solid foudation for researching the fuction of microRNA in CSFV infection.1. Two pairs of primers were designed to get CSFV 3'-UTR and the 5'-UTR gene. Through RT-PCR they have been successfully amplified from umbilical vein endothelial cells (SUVEC) infected with CSFV . After enzymatic assay, the fragment of 3'UTR and 5'UTR were recovered and was cloned into the psiCHECK?-2. The recombinant plasmid were exactly identified by sequencing and enzymatic digestion.2. According to miRBase, 76 microRNA has been found in swines. Useing RNA22 microRNA target detection software, these microRNA and CSFV3'-UTR were matched, of which four were found.3. The re-combinant plasmids and inhibitors of predicted microRNA (ssc-miR-let7c,ssc-miR-106a,ssc-miR-18,ssc-miR-139) were co-transfacted into SUVECs. The activity of Dual-Luciferase was detected by Luminometer. The CSFV3′-UTR is important site of microRNA in SUVECs. The four microRNAs mentioned above had some different impact on CSFV 3′-UTR, of which the ssc-miR-18 play a more important role.