Molecular characterization of genome segments 1 and 3 encoding two capsid proteins of Antheraea mylitta cytoplasmic polyhedrosis virus.

摘要: Antheraea mylitta cytoplasmic polyhedrosis virus (AmCPV), a cypovirus of Reoviridae family, infects Indian non-mulberry silkworm, mylitta, and contains 11 segmented double stranded RNA (S1-S11) in its genome. Some genome segments (S2 S6-S11) have been previously characterized but encoding viral capsid not characterized. In this study 1 (S1) 3 (S3) AmCPV were converted to cDNA, cloned sequenced. S1 consisted 3852 nucleotides, with one long ORF 3735 nucleotides could encode protein 1245 amino acids molecular mass ~141 kDa. Similarly, S3 3784 having 3630 1210 ~137 BLAST analysis showed 20-22% homology sequence spike proteins, respectively, other closely related cypoviruses like Bombyx mori CPV (BmCPV), Lymantria dispar (LdCPV), Dendrolimus punctatus (DpCPV). The ORFs expressed as 141 kDa 137 insoluble His-tagged fusion Escherichia coli M15 cells via pQE-30 vector, purified through Ni-NTA chromatography polyclonal antibodies raised. Immunoblot polyhedra, virion particles infected mid-gut the raised anti-p137 anti-p141 specific immunoreactive bands suggest that may code for structural proteins. Expression insect baculovirus recombinants produce (VLPs) by transmission electron microscopy. Immunogold staining encoded proteins self assembled form outer VLPs maintained their stability at different pH presence protein. Our results cloning, sequencing functional indicate can assemble remains associated it inner maintain stability. Further studies will help understand mechanism formation during replication.