The aim of the study was to identify the microRNAs that act on classical swine fever virus (CSFV) in swine umbilical vein endothelial cells (SUVECs). The 3'-and 5'-untranslated regions ( UTR) of CSFV were clon...The aim of the study was to identify the microRNAs that act on classical swine fever virus (CSFV) in swine umbilical vein endothelial cells (SUVECs). The 3'-and 5'-untranslated regions ( UTR) of CSFV were cloned and then inserted into psiCHECH TM-2 plasmids carrying Firefly and Renilla luciferases reporter genes; microRNAs that acted with CSFV were predicted by bio-informatics analysis; then the recombinant plasmids and inhibitors of the predicted microRNA were co-transfected into SUVECs. The activities of Firefly and Renilla luciferases were detected by luminometry. The PCR products of the CSFV 5'-UTR (373 bp) and 3'-UTR (252 bp) were detected by electrophoresis on 1% agarose gels. Four microRNAs (ssc-miR-let7c, ssc-miR-106a, ssc-miR-18, ssc-miR-139) were screened out and evaluated. The CSFV 3'-UTR is the important target site of microRNA in SUVECs. The four microRNAs mentioned above had different inhibitory effects on the CSFV 3'-UTR, of which the ssc-miR-18 played the most important role.展开更多
Classical swine fever virus(CSFV) is the causative agent of classical swine fever, a highly contagious disease of pigs. But there is little information on the recombination in natural populations of CSFVs. Therefore, ...Classical swine fever virus(CSFV) is the causative agent of classical swine fever, a highly contagious disease of pigs. But there is little information on the recombination in natural populations of CSFVs. Therefore, a phylogenetic analysis of 62 fulllength genome CSFV strains, isolated from all over the world, was performed to detect potential recombination events, with the recombinant sequences being analyzed with the SimPlot and RDP programs. The results identified a mosaic virus, Chinese CSFV HCLV(2)(AF091507.1), which is the one naturally emerged recombinant CSFV with two recombination breakpoints at 2 484 and 2 900 bp of the genome alignment. Its two putative parental-like strains were CSFV Shimen(AF092448.2) and CSFV strain C/HVRI(AY805221.1). This work demonstrated that homologous recombination did occur in natural CSFV populations. It had significant implications for understanding the molecular epidemiology of CSFV, and revealed that recombination was an important factor for high genetic diversities of CSFV.展开更多
文摘The aim of the study was to identify the microRNAs that act on classical swine fever virus (CSFV) in swine umbilical vein endothelial cells (SUVECs). The 3'-and 5'-untranslated regions ( UTR) of CSFV were cloned and then inserted into psiCHECH TM-2 plasmids carrying Firefly and Renilla luciferases reporter genes; microRNAs that acted with CSFV were predicted by bio-informatics analysis; then the recombinant plasmids and inhibitors of the predicted microRNA were co-transfected into SUVECs. The activities of Firefly and Renilla luciferases were detected by luminometry. The PCR products of the CSFV 5'-UTR (373 bp) and 3'-UTR (252 bp) were detected by electrophoresis on 1% agarose gels. Four microRNAs (ssc-miR-let7c, ssc-miR-106a, ssc-miR-18, ssc-miR-139) were screened out and evaluated. The CSFV 3'-UTR is the important target site of microRNA in SUVECs. The four microRNAs mentioned above had different inhibitory effects on the CSFV 3'-UTR, of which the ssc-miR-18 played the most important role.
基金Supported by the National Natural Science Foundation of China(31370140 31372438)
文摘Classical swine fever virus(CSFV) is the causative agent of classical swine fever, a highly contagious disease of pigs. But there is little information on the recombination in natural populations of CSFVs. Therefore, a phylogenetic analysis of 62 fulllength genome CSFV strains, isolated from all over the world, was performed to detect potential recombination events, with the recombinant sequences being analyzed with the SimPlot and RDP programs. The results identified a mosaic virus, Chinese CSFV HCLV(2)(AF091507.1), which is the one naturally emerged recombinant CSFV with two recombination breakpoints at 2 484 and 2 900 bp of the genome alignment. Its two putative parental-like strains were CSFV Shimen(AF092448.2) and CSFV strain C/HVRI(AY805221.1). This work demonstrated that homologous recombination did occur in natural CSFV populations. It had significant implications for understanding the molecular epidemiology of CSFV, and revealed that recombination was an important factor for high genetic diversities of CSFV.
