转录靶向猪瘟病毒3'-UTR shRNA细胞株的初步建立

Establishment of cell lines transcribing shRNA targeted to the 3'-UTR of CSFV

利用质粒载体在细胞内转录并加工成siRNA的方法,设计3对针对猪瘟病毒3'-UTR不同位点的干扰片段,分别与干扰载体pGenesil-1连接,转化DH5α,筛选阳性克隆得到重组干扰质粒pGene-1,pGene-2和pGene-3,脂质体介导转染重组干扰质粒于PK-15细胞,G418抗性筛选得到转录靶向猪瘟病毒3'-UTR shRNA的PK-15细胞株,为今后应用RNAi研究猪瘟病毒3'-UTR调控病毒复制的功能以及抑制猪瘟病毒增殖的研究奠定了基础。

Three pairs of interfering fragments targeted to 3＇-UTR of classic swine fever virus （CSFV） were designed and cloned into the plasmid pOenesil-1 respectively, generating recombinant plasmids pGene-1, pGene-2, pGene-3 respectively. These constructs were individually transformed into PK-15 cells by liposome. The cell lines transcribing shRNA for 3＇-UTR of CSFV were established by screening positive clones in the presence of G418. The established cell lines could serve as a foundation for further researches on the 3＇-UTR of CSFV and the inhibition of virus replication by RNAi.