摘要
针对O型口蹄疫病毒(Foot-and-mouth disease veirus,FMDV)基因组高度保守的VP1和3D区段,选取了120个干扰靶位,根据RNAi技术原理,构建了包括对照在内的82个shRNA表达质粒;通过细胞攻毒试验,提取细胞毒液RNA,然后RT-PCR,最后real-time PCR检测病毒和shRNA的表达量。结果显示,干扰质粒组基因表达抑制率为63%～96.8%,其中pSiRe-Zs-VP1-54和pSiRe-Zs-3D-12等2个表达质粒抑制效率最好,分别为96.8%和87%,表明这2个表达质粒能够明显抑制FMDV的复制。
One hundred twenty interference targets were selected aiming at the high conservative VPland 3D of O FMDV genome section. Based on the RNAi technological principles,82 shRNA expression plasmids including contrast were constructed. The cell venom RNA was collected from cell infection experiment,using RT-PCR,and Real- time PCR to test virus and the expressing level of shRNA. The results show that the inhibition ratio of gene expression of interference plasmids varied from 63% to 96.8% ,in which pSiRe-Zs-VP1-54 and pSiRe-Zs-3D-12 have the 96.8% and 87% of inhibition ratio, respectively. The research screened siNRA sequences have obvious inhibition from FMDV duplication at the cellular level using RNAi. It lays the foundation for transgenic clone of swine afterwords.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2012年第1期13-17,22,共6页
Chinese Journal of Veterinary Science
基金
转基因生物新品种培育国家重大专项(2009ZX08005-003B)
