1887

Abstract

In this study, we have deleted the gene of the baculovirus BmNPV, which encodes one of the viral RNA polymerase subunits, in order to create a knockout bacmid, Δ directing cytopathology-free single-cell infections for gene transduction and recombinant protein production. However, while removal of the complete ORF produced the expected phenotype, it also affected the function of the closely linked essential gene thus hampering the mutant bacmid rescue in cultured cells expressing recombinant LEF8. Subsequently, we determined that several diverse sequences can substitute for the 5′-upstream sequences that were removed by the deletion of the gene and also showed that neither a physical linkage nor expression of the two relevant genes under native promoter control is a prerequisite for a fully functional virus. Based on these findings, we generated a rescue-competent -null vector, which contained a heterologous promoter-driven This deficient vector, which produces productive infections and progeny virus lacking in deficiency-complementing cells expressing LEF8, could be used as the basis for an alternative to current silkmoth transduction systems.

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2016-03-01
2019-10-15
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