Expression of a foreign gene by cysteine proteinase null recombinant baculovirus

Baculovirus expression vector systems (BEVSs) are broadly used for producing foreign proteins in lepidopteran cells. Most commercial BEVSs are engineered to insert foreign genes into the polyhedrin (polh) locus. They lack the polh gene. These viruses cannot produce occlusion bodies and are inconvenient for per os inoculation of larvae. To avoid this, expression cassettes can be inserted in other parts of the virus genome. The preS2-S gene, coding for the recombinant middle surface antigen of the human hepatitis B virus (M-HBsAg), was expressed from the baculovirus construct rBmNPV-Δv-cath-M-HBsAg, inserting the foreign gene into the v-cath locus of Bombyx mori nucleopolyhedrovirus (BmNPV) so that v-cath was deleted and native polh was retained. Silkworm larvae were infected per os and M-HBsAg was observed to be abundantly produced till very late stages of infection. Infection of larvae with a mixture of the recombinant and wild-type baculoviruses was followed by degradation of the bulk of the produced M-HBsAg as early as 96 h after inoculation.