Baculovirus multigene expression vectors and their use for understanding the assembly process of architecturally complex virus particles.

摘要: Abstract The baculovirus expression vector is a eukaryotic DNA viral for the cloning and of foreign genes in cultured lepidopteran insect cells insects. It has become an important tool large-scale production recombinant proteins variety applications including structure-function analysis their gene products. We have developed number multigene vectors utilized these to understand assembly process multicomponent capsid structures large viruses such as bluetongue virus (BTV), member Orbivirus genus within family Reoviridae. BTV some 810 A diameter comprised two protein shells containing four major proteins, VP2, VP5, VP7 VP3, surrounding genome ten double-stranded RNA segments three minor (VP2, VP4 VP6). etiological agent sheep disease that sometimes fatal certain parts world (e.g., Africa, Asia, Americas). Using vectors, we co-expressed various combinations produced mimic stages assembly. For example, VP3 form core-like particles, while virus-like particles. deletion, point domain switching analyses each protein, been able identify sequences are essential These biochemical studies complemented by collaboration using cryoelectron microscopy image processing provide three-dimensional structure expressed In addition with other associates, used X-ray crystallography deduce its atomic structure. Extensive on immune responses elicited self-assembled chimeric derivatives involving antigens, carried out. Finally, little 10 μg shown they can confer long-lasting protection against BTV.