Abstract：N-linked glycosylation site is critical for the infection and proliferation of most virus, elimination or inhibition of glycosylation is becoming a potent therapy of some virus such as Dengue virus. To elucidated roles of N-linked glycosylation in the infectivity and duplication of Japanese encephalitis virus (JEV), in this study, endo-glycosidase enzymes peptide N-glycosidase F (PNGase F) and endoglycosidase H (Endo H) were used to digest JEV to remove the the N-glycosylation on the virus particles surface. Western blotting analysis showed that the glycan chains of JEV particles was successfully removed. To observe the changes in JEV infectivity and proliferation, plaque assay was performed using treated JEV on BHK-21 cells. In another exprement, BHK-21 cells were treated with tunicamycin and infected with JEV. The results showed that JEV digested by PNGase F or Endo H formed smaller plaques than that of control group, while JEV titer was reduced significantly. The plaques formed by JEV in cells treated with tunicamycin were not observed significant differences in size, but JEV titer was also reduced significantly. From these results we conformed that N-glycosylation plays an important role in the process of JEV infection and proliferation, without N-glycosylation, the infectivity and proliferative ability of JEV will be reduced remarkably.This study arises a new potential strategy on JEV therapy.
