Inhibition Of Gene Expression Of HCMV UL49 By External Guide Sequence DNA

RNaseP is a ribonucleoprotein complex within most kinds of cells, and catalyzes a hydrolysis reaction to remove the leader sequence of precursor tRNA. Studies show that RNaseP recognize the structure rather than the primary nucleotide sequence of the substrates, and can cleave a model substrate that contains two important parts:NCCA-3' terminal and RNA helix area. External Guide Sequence(EGS) is synthesized to combine with mRNA to form the structure similar to ptRNA, and guide RNaseP to cleave the mRNA, so the gene expression was inhibited.We designed DNA-based EGS with the targeted sequence of HCMVUL49 mRNA, study the inhibition of gene expression of HCMVUL49with DNA-based EGS .pEGFP-N1 is a kind vector which expressed in eukaryota . HCMVUL49 gene was cloned to pEGFP-N1 (pEGFP-UL49) , and coexpressed with EGFP' N terminal . DNA-based EGS and pEGFP-UL49 were cotransfected into the cells of hela, and cultured 24 hours. We can see the expression level of HCMVUL49 gene by detecting the fluorescence with microscope and Flow Cytometer; and detecting the mRNA directly to know the cleavage efficiency with real-time PCR.The results show that gene expression of HCMVUL49 was inhibited by DNA-based EGS we designed, allowed it to be further applied in antivirus study. And our study also reveal the potential of DNA-based EGS .