Abstract:
:DNA polymerase (DNApol) is highly conserved in baculovirus and is required for viral DNA replication. However, little is known about gammabaculovirus DNApol. Here DNApol of the gammabaculovirus Neodiprion lecontei nucleopolyhedrovirus (NeleNPV) was cloned into a dnapol-null alphabaculovirus AcMNPV bacmid, creating Bac-GFP-AcΔPol-NlPol. The resulting recombinant bacmid did not spread to neighboring cells, virus growth curve and real-time PCR revealed that NeleNPV dnapol substitution did not rescue AcMNPV DNA replication and virus production. Immunofluorescence microscopy revealed that NeleNPV DNApol was expressed but could not localize to the nucleus. Subsequently NeleNPV DNApol was fused to SpltNPV DNApol nuclear localization signal (NLS) and the fused DNApol could import into nucleus. The NLS-fusing NeleNPV DNApol was further transposed into the dnapol-null AcMNPV bacmid, creating Bac-GFP-AcΔPol-HA:NlPolNLS. The recombinant virus could replicate and produce infectious virus in Sf9 cells, albeit at reduced levels compared to wild type AcMNPV. Taken together, our results suggested that the NLS deficiency of NeleNPV DNApol blocked viral DNA replication and production of infectious virus in dnapol-null AcMNPV bacmid.
journal_name
Virus Resjournal_title
Virus researchauthors
Chen G,Fang Y,Yan Q,Li P,Wu L,Feng Gdoi
10.1016/j.virusres.2019.04.005subject
Has Abstractpub_date
2019-06-01 00:00:00pages
52-57eissn
0168-1702issn
1872-7492pii
S0168-1702(19)30057-7journal_volume
266pub_type
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