Characterization of a late gene, ORF75 from Bombyx mori nucleopolyhedrovirus
Open reading frame 75 (Bm-p33) of Bombyx mori nucleopolyhedrovirus (BmNPV) is a homologue of Autographa californica multiple nucleopolyhedrovirus ORF92. The gene is conserved among all baculoviruses that have been completely sequenced to date and is considered to be a baculovirus core set gene. No amino acid mutation was found in Bm-p33 sequences among six BmNPV strains differing in geography, phenotype, or host. The Bm-p33 transcript can be detected as early as 12 h post infection (h p.i.) and remains detectable until 96 h p.i. The Bm-p33 protein was detected in cell lysates from 18 h p.i. through 96 h p.i., and no positive band could be detected in budded viruses (BVs) and occlusion-derived viruses (ODVs) by western blot using anti-Bm-p33 serum. Immunofluorescence microscopy indicated that Bm-p33 accumulated in the nuclear membrane and the intranuclear region, especially near the nuclear membrane of the virus-infected cells. Bm75 RNAi significantly decreased the mRNA level. However, no obvious effects on ODV formation and BV production in BmNPV-infected cells could be detected. Bm-p33 is a BmNPV late gene encoding a nonstructural protein which may function mainly in the nucleus of the infected cells.
KeywordsBombyx mori nucleopolyhedrovirus Bm75 P33 Late gene Nonstructural protein
This project was supported by the National Natural Science Foundation of China (30570074).
- 8.Zhang X, Shen W, Lu Y et al (2010) Expression of UreB and HspA of Helicobacter pylori in silkworm pupae and identification of its immunogenicity. Mol Biol Rep. doi: 10.1007/s11033-010-9988-2
- 21.Gong CL, Lu KM (1993) Studies on Bombyx mori nuclear polyhedrosis virus with tetragonal polyhedron. Sci Seric 19:25–31Google Scholar
- 28.Ohkawa T, Washburn JO, Sitapara R et al (2005) Specific binding of Autographa californica M nucleopolyhedrovirus occlusion-derived virus to midgut cells of Heliothis virescens larvae is mediated by products of pif genes Ac119 and Ac022 but not by Ac115. J Virol 79:15258–15264PubMedCrossRefGoogle Scholar