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KMID : 0380220060390060737
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Journal of Biochemistry and Molecular Biology
2006 Volume.39 No. 6 p.737 ~ p.742
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Characterization of a Late Gene, ORF60 from Bombyx mori Nucleopolyhedrovirus
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Du Meng-Fang
Yin Xin-Ming
Guo Zhong-Jian
Zhu Liang-Jun
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Abstract
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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.
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KEYWORD
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BmNPV, Bm60, Expression, RT-PCR, Subcellular location
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