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Journal of General Virology header logo Journal of General Virology

Volume 79, Issue 8

Isolation and characterization of a temperature-sensitive mutant of Bombyx mori nucleopolyhedrovirus for a putative RNA polymerase gene No Access

Abstract

Temperature-sensitive (ts) mutants of nucleopolyhedrovirus (BmNPV), which exhibited no polyhedra formation at the non-permissive temperature of 33 °C, wereproduced using the base analogue 5-bromodeoxyuridine. A unique ts mutant, designated ts-S1, was characterized in terms of gene mutation and virulence in cultured cells and silkworm larvae. Mutant-infected BmN4 cells at 33 °C showed normal viral DNA synthesis but defective budded virus production and polyhedrin synthesis, suggesting the absence of late and very late gene expression. Silkworm larvae were injected with ts-S1 and reared at 33·5 °C. At 7 days postinjection, none of the larvae exhibited nucleopoly- hedrosis but some possessed viral DNA, detected by PCR using virus-specific primers. Continued rearing of the larvae at a permissive temperature of 25°C showed that, while most developed into normal adults, some developed nucleopolyhedrosis, indicating that the former larvae had aborted the virus infection during the course of rearing at 33·5°C. No viral DNA was detected in the adults. Marker rescue tests to identify the lesion involved in the ts phenotype of ts-S1, and nucleotide sequencing of the identified genome region, showed a single nucleotide mutation of a putative RNA polymerase gene, late expression factor-8 These results indicate that is essential for BmNPV replication and

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1998-08-01
2025-12-09

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/content/journal/jgv/10.1099/0022-1317-79-8-2071
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Isolation and characterization of a temperature-sensitive mutant of Bombyx mori nucleopolyhedrovirus for a putative RNA polymerase gene
J. Gen. Virol. 79, 2071 (1998); https://doi.org/10.1099/0022-1317-79-8-2071
/content/journal/jgv/10.1099/0022-1317-79-8-2071
/content/journal/jgv/10.1099/0022-1317-79-8-2071
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