Objective: To establish stable expressing system of Borna disease virus (BDV) phosphoprotein in PC-12 cells, and then study its influence on cell proliferation of PC-12 cells.
Method: An expression plasmid with green fluorescence protein was cloned and identified to express BDV phosphoprotein. Cultured PC-12 cell was transfected with the recombinant plasmid by positive ion lipidsome method. Fluorescence microscopy was used to detect the expression of phosphoprotein in PC-12 cells, then G418 was added into cell culture medium to kill cells without recombinant plasmid. We performed reverse transcriptase polymerase chain reaction (RT-PCR) in the 10th generation of treated cells to examine the expression of BDV phosphoprotein. The proliferation of treated cells and control cells was examined by methyl thiazolyl tetrazolium assay (MT).
Result: The recombinant plasmid was confirmed to be able to express BDV phosphoprotein and green fluorescence protein by both fluorescence microscopy and RT-PCR. BDV phosphoprotein expressed in PC-12 cell inhibited cell proliferation.
Conclusion: We established a stable expressing system of BDV phosphoprotein in PC-12 cell. This cell model can be used to study the effect of BDV phosphoprotein on the centre nervous system without exposure to live virus.