Expression Plasmid And Interfering Plasmid Construction Of Severe Hepatitis Related Gene Hfgl2 And Its RNA Interference In Vitro

Objective In order to explore the opportunity of RNA interference technique in the inhibitory application of hfgl2 expression, which has been reported to be involved in a variety of disease development including fulminant viral hepatitis and acute rejection of allo/xero transplantation, fetal loss syndrome.Methods hfgl2shRNA expression plasmids were constructed. pcDNA3.1-hfgl2 was constructed to show hfgl2 protein expression. pEGFP-hfgl2 expressing hfgl2-EGFP fusion protein was constructed for the screening of the effect of hfgl2shRNA expression plasmids on the hfgl2 expression. The plasmid of irrelevant protein expression of pEGFP and a construct named irrelevant shRNA with a random combination of the p-hfgl2shRNA sequence were used as controls. The inhibibion of hfgl2 expression by hfgl2shRNA was analyzed by direct observation through fluorescent microscopy.Results The experiments showed the significant inhibitory effect of p-hfgl2shRNA1 and p-hfgl2shRNA2 on the hfgl2 expression at 24h and 48h post-cotransfection by observation of green fluorescent cells in CHO cell line. Conclusions The study demonstrated that the construct of p-hfgl2shRNA successfully interfered the hfgl2 expression in vitro and it provides the foundation for further investigation of this constructs'application in vivo and further more as a therapeutic strategy for a targeting intervention in the diseases control which fgl2 gene contributes to.