{Reference Type}: Journal Article
{Title}: Generation of Complex Protein Structures by Coinfection with High-Quality Recombinant Baculovirus.
{Author}: Walker S;Rose J;Hayes BC;Mulvania T;
{Journal}: Methods Mol Biol
{Volume}: 2829
{Issue}: 0
{Year}: 2024
暂无{DOI}: 10.1007/978-1-0716-3961-0_10
{Abstract}: The baculovirus expression vector system (BEVS) is a powerful platform for protein expression in insect cells. A prevalent application is the expression of complex protein structures consisting of multiple, interacting proteins. Coinfection with multiple baculoviruses allows for production of complex structures, facilitating structure-function studies, allowing augmentation of insect cell functionality, and production of clinically relevant products such as virus-like particles (VLPs) and adeno-associated viral vectors (AAV). Successful coinfections require the generation of robust and well-quantified recombinant baculovirus stocks. Virus production through homologous recombination, combined with rigorous quantification of viral titers, allows for synchronous coinfections producing high end-product titers. In this chapter, we describe the streamlined workflow for generation and quantification of high-quality recombinant baculovirus stocks and successful coinfection as defined by a preponderance of dually infected cells in the insect cell culture.