1887

Abstract

During cell infection, the gene of baculoviruses frequently mutates, producing the few polyhedra (FP) per cell phenotype with reduced polyhedrin () expression levels compared with wild-type baculoviruses. Here we report that the gene of the model baculovirus, multiple nucleopolyhedrovirus (AcMNPV), contains two hypermutable seven-adenine (A7) mononucleotide repeats (MNRs) that were mutated to A8 MNRs and a TTAA site that had host DNA insertions, producing mutants during 21 cell infection. The FP phenotype in 9 and Hi5 cells was more pronounced than in 21 cells. AcMNPV mutants produced similar levels of polyhedra or enhanced GFP, which were both under the control of the AcMNPV promoter for expression, in 21 cells but lower levels in 9 and Hi5 cells compared with AcMNPV with an intact gene. This correlated with the mRNA levels detected in each cell line. The majority of 21 cells infected with mutants showed high promoter-mediated GFP expression levels. Two cell lines subcloned from 21 cells that were infected with mutants showed different GFP expression levels. Furthermore, a small proportion of Hi5 cells infected with mutants showed higher production of polyhedra and GFP expression than the rest, and the latter was not correlated with increased m.o.i. Therefore, these data suggest that AcMNPV promoter-mediated gene expression activities differ in the three cell lines and are influenced by different cells within the cell line.

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2013-01-01
2019-10-21
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