A novel expression system based on host-range expansion of baculovirus

Area of Science:

• Molecular Biology
• Virology
• Biotechnology

Background:

• Autographa californica multiple-nucleocapsid nucleopolyhedrosis virus (AcMNPV) is a widely used baculovirus expression vector.
• Bombyx mori nuclear polyhedrosis virus (BmNPV) has a distinct host range and genomic characteristics.
• Expanding the host range of baculoviruses can enhance their utility for foreign gene expression.

Purpose of the Study:

• To develop a recombinant baculovirus with an expanded host range for expression in Bombyx mori.
• To construct a novel expression vector system for efficient foreign gene production in silkworms.
• To evaluate the feasibility of using the engineered virus for mass production of recombinant proteins.

Main Methods:

• Cotransfection of bacmid DNA with a BmNPV HindIII M fragment to create AcMNPV/r2.
• Construction of a recombinant transposon vector containing a mutant green fluorescent protein (GFP) gene and a polyhedrin gene.
• Generation of recombinant viruses (AcMNPV/r3 and AcMNPV/r4) through transfection into Sf-9 cells and subsequent infection of Bombyx mori cells and larvae.

Main Results:

• A host range expanded recombinant virus, AcMNPV/r4, was successfully generated.
• High expression of the green fluorescent protein (GFP) was observed in Bombyx mori larvae infected with AcMNPV/r4.
• The developed expression system demonstrated practicality for foreign gene product mass production.

Conclusions:

• The engineered recombinant baculovirus exhibits an expanded host range, enabling expression in Bombyx mori.
• This novel expression system is effective for high-level production of foreign gene products in silkworms.
• The findings support the application of this system for the industrial-scale manufacturing of valuable recombinant proteins.