猪繁殖与呼吸综合征病毒相关猪microRNA的初步筛选  

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

作　　者：郜原[1] 薄联锋[1] 武小椿[1] 孙柏[1] 温俊歌[1] 孙岩[1] 徐志坤[1] 张德礼[1] 

机构地区：[1]西北农林科技大学动物医学院兽医微生物学与病毒学课程组病毒免疫与生物信息研究室,陕西杨凌712100

出　　处：《中国兽医杂志》2012年第3期21-24,共4页Chinese Journal of Veterinary Medicine

基　　金：国家自然科学基金面上项目(31072115)

摘　　要：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。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.

关 键 词：猪繁殖与呼吸综合征病毒(PRRSV) microRNA(miRNA) 初步筛选 

分 类 号：S852.659.6[农业科学—基础兽医学]