猪繁殖与呼吸综合征病毒相关猪microRNA的初步筛选

本试验旨在初步筛选出与猪繁殖与呼吸综合征病毒(PRRSV)感染相关的猪microRNA(miRNA)。采用生物信息学方法预测可能与PRRSV 3′非编码区(3′utr)产生相互作用的猪miRNA,构建PRRSV 3′utr的双荧光素酶报告基因载体psi-CHECK-utr,同时合成预测到的miRNA(ssc-miR-323、ssc-miR-105-1)及其相应的抑制物,共转染猪脐静脉血管内皮细胞系(SUVEC),检测双荧光素酶活性。结果发现,ssc-miR-323与psiCHECK-utr共转染后,荧光比率略有升高,ssc-miR-105-1与psiCHECK-utr共转染后,荧光比率降低。初步判定ssc-miR-323对PRRSV 3′utr没有抑制作用,而ssc-miR-105-1对PRRSV3′utr有一定的抑制作用。

Primary selection of microRNA interacting with porcine reproductive and respiratory syndrome virus

The aim of the study was to filtrate the miRNA which interact with porcine reproductive and respiratory syndrome virus(PRRSV).We inserted the PRRSV 3′-untranslated region(3′utr) into psiCHECHTM-2 plasmid and constructed the recombinant plasmid psiCHECK-utr which carrying Firefly and Renilla luciferases reporter genes.The swine miRNAs(ssc-miR-323 and ssc-miR-105-1) which may target PRRSV 3′utr were predicted by the computer.And then we synthesized the miRNAs and their inhibitors,co-transfected the luciferase vector with miRNAs or inhibitors into swine umbilical vein endot helial cells(SUVECs).Meanwhile,we detected the luciferase activity by the luminometer and analyzed the relative by computer.The results indicated that ssc-miR-323 has no effect on PRRSV 3′utr expression,whereas ssc-miR-105-1 could inhibit the PRRSV 3′utr to some extent.