Baculovirus Expression Strategies for Protein Complex Production

Abstract Protein complexes often contain multiple subunits, and it is these complexes that are at the core of biological function in the cell. Inherently low quantities of protein complexes in eukaryotes can prohibit the extraction of these essential cellular machines from native source material, posing a considerable challenge for functional analysis at the molecular level. The baculovirus expression vector system (BEVS) is particularly useful for recombinantly producing eukaryotic proteins in the quality and quantity required for deciphering their structure and function. Recent efforts have focused on improving reagents and streamlining protocols to routinely express protein complexes by recombinant baculoviruses. The challenge of integration of baculovirus expression in high‐throughput production pipelines for proteins and their complexes is being addressed. Key Concepts: Protein complexes are a cornerstone of biological function in the cell. Recombinant production is essential to study many multiprotein complexes. Ongoing development of expression systems for producing multiprotein complexes. BEVS as a method of choice for large‐scale production of complexes requires integration of all genes in a single multigene baculovirus rather than coinfecting insect cell cultures with many baculoviruses. Creation of new reagents and streamlined protocols to address the challenge of generating multigene baculoviruses for routine laboratory use. Engineering baculoviruses and new host cell lines to further enhance protein production by BEVS. Multigene vector generation incorporated into automation pipelines to increase the throughput of protein complex expression by BEVS.