Immobilization of foreign protein in BmNPV polyhedra by fusion expression with partial polyhedrin fragments

Abstract

Bombyx mori nucleopolyhedrovirus (BmNPV) produces large, proteinaceous crystal matrix named polyhedra, which occlude progeny virions which are produced during infection and protect virions from hostile environmental conditions. In this study, five overlapping N-terminal fragments of the BmNPV polyhedrin ORF were cloned and ligated with the foreign gene egfp, and five recombinant baculoviruses were constructed by BmNPV(Polh(+)) Bac-to-Bac baculovirus expression system was used to co-express the polyhedrin and fused protein. The results showed that the fusion proteins were highly expressed, and the foreign proteins fused with the 100aa fragment of polyhedrin could be embedded into polyhedra at a higher ratio. This study provides a new method for efficient preservation of useful proteins for the development of new biopesticide with toxin protein and delivery vector system of vaccines.

Keywords: Bombyx mori nucleopolyhedrovirus; Polyhedra; Polyhedrin; Protein immobilization.