NDLI: Bombyx mori nucleopolyhedrovirus lef8 gene: effects of deletion and implications for gene transduction applications.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Ioannidis, Konstantinos Swevers, Luc Iatrou, Kostas
Description	Country affiliation: Greece Author Affiliation: Ioannidis K ( Insect Molecular Genetics and Biotechnology Group, Institute of Biosciences and Applications, National Centre for Scientific Research 'Demokritos', 15310 Aghia Paraskevi, Athens, Greece.); Swevers L ( Insect Molecular Genetics and Biotechnology Group, Institute of Biosciences and Applications, National Centre for Scientific Research 'Demokritos', 15310 Aghia Paraskevi, Athens, Greece.); Iatrou K ( Insect Molecular Genetics and Biotechnology Group, Institute of Biosciences and Applications, National Centre for Scientific Research 'Demokritos', 15310 Aghia Paraskevi, Athens, Greece.)
Abstract	In this study, we have deleted the lef8 gene of the baculovirus BmNPV, which encodes one of the viral RNA polymerase subunits, in order to create a knockout bacmid, Δlef8, directing cytopathology-free single-cell infections for gene transduction and recombinant protein production. However, while removal of the complete lef8 ORF produced the expected phenotype, it also affected the function of the closely linked essential gene orf40, thus hampering the mutant bacmid rescue in cultured Bombyx cells expressing recombinant LEF8. Subsequently, we determined that several diverse sequences can substitute for the orf40 5'-upstream sequences that were removed by the deletion of the lef8 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 lef8-null vector, which contained a heterologous promoter-driven orf40. This lef8-deficient vector, which produces productive infections and progeny virus lacking lef8 in deficiency-complementing cells expressing LEF8, could be used as the basis for an alternative to current silkmoth transduction systems.
File Format	HTM / HTML
ISSN	00221317
Issue Number	3
Volume Number	97
e-ISSN	14652099
Journal	Journal of General Virology
Language	English
Publisher	Microbiology Society
Publisher Date	2016-03-01
Publisher Place	Great Britain (UK)
Access Restriction	Subscribed
Subject Keyword	Nucleopolyhedrovirus Research Support, Non-u.s. Gov't Transduction, Genetic Discipline Virology Viral Proteins Dna-directed Rna Polymerases Metabolism Bombyx Journal Article Virology Animals Gene Expression Regulation, Viral Gene Deletion Genetics
Content Type	Text
Resource Type	Article
Subject	Virology

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