info:eu-repo/semantics/article
A simplified strategy to package foreign proteins into baculovirus occlusion bodies without engineering the viral genome
Fecha
2020-01-10Registro en:
Fabre, Maria Laura; Masson, Tomas; Haase, Santiago; Ferrelli, Maria Leticia; Romanowski, Victor; A simplified strategy to package foreign proteins into baculovirus occlusion bodies without engineering the viral genome; Elsevier Science; Journal of Biotechnology; 307; 10-1-2020; 175-181
0168-1656
CONICET Digital
CONICET
Autor
Fabre, Maria Laura
Masson, Tomas
Haase, Santiago
Ferrelli, Maria Leticia
Romanowski, Victor
Resumen
Polyhedron envelope protein (PEP) is the major component of the calyx that surrounds the baculovirus occlusion body (OB). PEP has been associated with the stabilization and resistance of polyhedra in the environment. Due to the abundant levels of PEP in OBs, we decided to use this protein as a fusion partner to redirect foreign proteins to baculovirus polyhedra. In this study we developed a strategy that involves the generation of a monoclonal transformed insect cell line expressing a protein of interest fused to the the Anticarsia gemmatalis multiple nucleopolyhedrovirus (AgMNPV) N-terminus of PEP that enables the packaging of foreign proteins into the OBs without generating a recombinant baculovirus. This proved to be an efficient platform that could be exploited to improve wild type baculovirus for their use as bioinsecticides without facing the concerns of releasing genetically modified DNA to the environment and bypassing the associated regulatory issues. We demonstrated, using immunological, proteomic and microscopy techniques, that the envelope of AgMNPV OBs can effectively trap chimeric proteins in an infected insect cell line expressing AgMNPV PEP fused to the enhanced green fluorescent protein (eGFP). Furthermore, packaging of chimeric PEP also took place with heterologous OBs such as those of Autographa californica multiple nucleopolyhedrovirus (AcMNPV), another group I alphabaculovirus.