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.
Ge
Jun-Qing
JQ
Institute of Insect Sciences and Ministry of Agriculture Key Laboratory of Molecular Biology of Crop Pathogens and Insects, Zhejiang University, Kaixuan Road 268, Hangzhou 310029, People's Republic of China.
Gao
Guo-Hui
GH
Xu
Yi-Peng
YP
Zhang
Chuan-Xi
CX
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D012831:Bombyx / Q000821:virology*; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D005455:Fluorescent Antibody Technique; D015967:Gene Expression Regulation, Viral; D005814:Genes, Viral / Q000235:genetics*; D008969:Molecular Sequence Data; D017924:Nucleopolyhedroviruses / Q000235:genetics* / Q000254:growth & development; D010802:Phylogeny; D021381:Protein Transport; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D013997:Time Factors; D014158:Transcription, Genetic; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
D012333:RNA, Messenger; D014764:Viral Proteins