Characterization of a late gene, ORF60 from Bombyx mori nucleopolyhedrovirus.
Open reading frame 60 of Bombyx mori nucleopolyhedrovirus (Bm60) is located between 56,673 and 57,479 bp in the BmNPV genome which encodes 268 amino acid residues with predicted molecular weight of 31.0 kDa. Bm60 and its homologues have been identified in 11 completely sequenced lepidopteran NPVs. The transcript of Bm60 was detected by RT-PCR at 18-72 h post-infection (p.i.), while the corresponding protein could be detected at 24-72 h p.i. in BmNPV-infected BmN cells by Western blot analysis using a polyclonal antibody against Bm60. The expression of Bm60 was inhibited in the presence of Ara-c, an inhibitor of viral DNA synthesis. These results together indicated that Bm60 was a late gene. The size of Bm60 product was found to be a 31 kDa in BmNPV-infected BmN cells, consistent with predicted molecular weight. Immunofluoresence analysis showed that the Bm60 product was first detected in the cytoplasm at 24 h p.i and also located in nucleus during later infection. In conclusion, the available data suggest that Bm60 is a functional ORF of BmNPV and encodes a 31kDa protein expressed in the later stage of infection cycle.
Du
Meng-Fang
MF
College of Animal Science, Zhejiang University, Hangzhou, 310029, China.
Yin
Xin-Ming
XM
Guo
Zhong-Jian
ZJ
Zhu
Liang-Jun
LJ
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D012831:Bombyx / Q000821:virology*; D003001:Cloning, Molecular; D003561:Cytarabine / Q000494:pharmacology; D015870:Gene Expression; D005814:Genes, Viral; D007118:Immunoassay; D008969:Molecular Sequence Data; D017924:Nucleopolyhedroviruses / Q000235:genetics*; D016366:Open Reading Frames; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D013347:Subcellular Fractions; D014764:Viral Proteins / Q000235:genetics* / Q000378:metabolism
D014764:Viral Proteins; D003561:Cytarabine