Biochemical and Biophysical Research Communications
Expression profiling of baculovirus genes in permissive and nonpermissive cell lines
Section snippets
Materials and methods
Cell lines, viruses, plaque assay, and virus growth curves. The Sf-9 and BmN cell lines were maintained as described previously on TC-100 medium supplemented with 10% fetal bovine serum [21]. The AcMNPV E2 [24] isolate was propagated in Sf-9 cells. The BmNPV T3 [25] and eh2-AcMNPV [12] isolates were propagated in BmN cells. Virus growth curves were determined by plaque assay as described previously [26], [27]. Titration of eh2-AcMNPV was performed on BmN cells.
Microarray analysis. Microarray
Gene expression patterns in Sf-9 and BmN cells infected with AcMNPV or BmNPV
As reported previously, BmNPV and AcMNPV are highly homologous at DNA sequence and genome organization levels, but their host ranges are nonoverlapping [10], [11]. In our hands, BmNPV strongly replicated (titer of >108 pfu/ml at 48 h p.i.) and produced PIBs in the B. mori-derived BmN cell line, but it replicated very weakly in the Spodoptera frugiperda-derived Sf-9 cell line (Figs. 1A and B). In contrast, AcMNPV strongly replicated (titer of about 108 pfu/ml at 48 h p.i.) and produced PIBs in Sf-9
Acknowledgments
This work was supported in part by grants from BRAIN (to M.K.) and JSPS (Nos. 14360032 and 14656023) (to T.S.), Japan.
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These three authors contributed equally to this work.
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Present address: New Frontiers Research Laboratory, Toray Industries, Inc., Tebiro 1111, Kamakura, Kanagawa 248-8555, Japan.