title: Phenotypic grouping of 141 BmNPVs lacking viral gene sequences.
abstract: We constructed a series of gene knockout BmNPVs (KOVs) for each of 141 genes (Gomi et al., 1999; Katsuma et al., 2011) using the BmNPV T3 bacmid system (Ono et al., 2007) and lambda red recombination system (Datsenko and Wanner, 2000). In a subsequent analysis of the properties needed for infection using a marker gene, egfp (enhanced green fluorescent protein gene), inserted into the polyhedrin locus, the knockout viruses (KOVs) were subdivided into four phenotypic types, A to D. Type-A (86 KOVs) showed the ability to expand infections equivalent to the control while type-B (8 KOVs) spread infections more slowly. Type-C (37 KOVs) expressed egfp in transfected-BmN cells but the production of infectious viruses was not observed. Type-D (10 KOVs) showed no ability to express egfp even in the transfection experiments. KOVs lacking genes (pkip (Bm15), gp41 (Bm66), bro-d (Bm131), Bm20, 48, 65, 91, 93, or 101) previously identified as being essential, were placed in the viable type-A and B categories.
authors.affiliation: Laboratory of Applied Molecular Entomology, Division of Applied Bioscience, Graduate School of Agriculture, Hokkaido University, Sapporo 060-8589, Japan.
authors.forename: Chikako
authors.initials: C
authors.lastname: Ono
authors.forename: Takanori
authors.initials: T
authors.lastname: Kamagata
authors.forename: Hitomi
authors.initials: H
authors.lastname: Taka
authors.forename: Ken
authors.initials: K
authors.lastname: Sahara
authors.forename: Shin-ichiro
authors.initials: S
authors.lastname: Asano
authors.forename: Hisanori
authors.initials: H
authors.lastname: Bando
mesh_terms: D000818:Animals; D012831:Bombyx; D002460:Cell Line; D017353:Gene Deletion; D017930:Genes, Reporter; D005814:Genes, Viral; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D050296:Microbial Viability; D017924:Nucleopolyhedroviruses / Q000235:genetics / Q000502:physiology*; D013194:Staining and Labeling / Q000379:methods; D014779:Virus Replication
chemical_list: C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins