An improved method for the rescue of recombinant Newcastle disease virus

Abstract

Reverse genetics has been used to generaterecombinantNewcastledisease virus with enhanced immunogenic properties for vaccine development. The system, which involves co-transfecting the viral antigenomic plasmid with three helperplasmidsintoaT7RNApolymeraseexpressingcelltoproduceviralprogenies, posesagreatchallenge.Wehavemodified the standard transfection method to improve the transfection efficiency of the plasmids, resulting in a higher titer of virus progeny production. Two transfection reagents (i.e., Lipofectamine and polyethylenimine) were used to compare the transfection efficiency of the four plasmids. The virus progenies produced were quantitated with flow cytometry analysis of the infectious virus unit. The modified transfection method increased thetiterofvirusprogeniescomparedwith thatofthestandardtransfectionmethod.