Demonstration of the protective effects of fluorescent proteins in baculoviruses exposed to ultraviolet light inactivation

Autographa californica multiple nucleopolyhedrovirus (AcMNPV) recombinants, namely AcRFP produced by fusion of the red fluorescent protein (RFP) gene with the polyhedrin gene, and a recombinant (pAcUW21-23GFP) carrying the green fluorescent protein (GFP) in its viral envelope, were evaluated for their resistance to inactivation by ultraviolet light. AcRFP recombinants produced incomplete polyhedra with low infectivity for Trichoplusia ni larvae, whereas AcuW21-23GFP produced normal polyhedra with high infectivity. Electron microscopy of AcRFP CL14 showed the incorporation of very few viral particles into polyhedrin matrix protein material. The LC(50) for AcuW21-23GFP was 0.10 occlusion bodies/mm(2), whereas the LC(50) values for several AcRFP recombinants ranged from 20 to 329 occlusion bodies/mm(2). When both the RFP and GFP recombinants were exposed to ultraviolet light (UV-B 280–320 nm), the results support the conclusion that these fluorescent proteins afford some protection against its damaging effects. Abbreviation: / AcMNPV: Autographa californica multiple nucleopolyhedrovirus BV: budded virus CPE: cytopathogenic effect ECV: extracellular virus OB: occlusion body ODV: occlusion derived virus RFP: red fluorescent protein GFP: green fluorescent protein TCID(50): tissue culture infective dose at the 50 % level UV-B: ultraviolet light of 280–320 nm