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|標題:||Autographa californica Multiple Nucleopolyhedrovirus LEF-2 Is a Capsid Protein Required for Amplification but Not Initiation of Viral DNA Replication||作者:||Wu, C.P.
|關鍵字:||nuclear polyhedrosis-virus;late gene-expression;helicase-primase;complex;mammalian-cells;baculovirus dna;early promoter;identification;infection;localization;ie-1||Project:||Journal of Virology||期刊/報告no：:||Journal of Virology, Volume 84, Issue 10, Page(s) 5015-5024.||摘要:||
The late expression factor 2 gene (lef-2) of baculovirus Autographa californica multiple nucleopolyhedrovirus (AcMNPV) has been identified as one of the factors essential for origin-dependent DNA replication in transient expression assays and has been shown to be involved in late/very late gene expression. To study the function of lef-2 in the life cycle of AcMNPV, lef-2 knockout and repair bacmids were generated by homologous recombination in Escherichia coli. Growth curve analysis showed that lef-2 was essential for virus production. Interestingly, a DNA replication assay indicated that lef-2 is not required for the initiation of viral DNA replication and that, rather, it is required for the amplification of DNA replication. lef-2 is also required for the expression of late and very late genes, as the expression of these genes was abolished by lef-2 deletion. Temporal and spatial distributions of LEF-2 protein in infected cells were also analyzed, and the data showed that LEF-2 protein was localized to the virogenic stroma in the nuclei of the infected cells. Analysis of purified virus particles revealed that LEF-2 is a viral protein component of both budded and occlusion-derived virions, predominantly in the nucleocapsids of the virus particles. This observation suggests that LEF-2 may be required immediately after virus entry into host cells for efficient viral DNA replication.
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