1887

Abstract

Open reading frame (ORF) 67 () of the nucleopolyhedrovirus (BmNPV) is a highly conserved gene that is found in all completely sequenced baculoviruses; its function is unknown. In the present study, a -knockout virus was generated for studying the role of Bm67 in the BmNPV infection cycle. Furthermore, a -repair bacmid was constructed by transposing the native promoter-promoted ORF into the locus of the -knockout bacmid. After these recombinant bacmids were transfected into BmN cells, the -knockout bacmid caused defects in the production of infectious budded viruses. However, the -repair bacmid could rescue the defect, and budded virus titres reached wild-type levels. Quantitative real-time PCR analysis indicated that Bm67 is required for normal levels of DNA synthesis or for the stability of nascent viral DNA at the early stage. Electron microscopic analysis revealed that the formation of normal-appearing nucleocapsids is reduced in -knockout bacmid-transfected cells, and nucleocapsids are rarely found in the cytoplasm. The presence of ‘enveloped’ nucleocapsids at the nucleoplasm bilayer indicated that they are enveloped abnormally. These results indicated that Bm67 is required for the production of infectious budded viruses and for assembly of envelope and nucleocapsids.

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2008-03-01
2024-12-08
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