Skip to main content
Journal of Virology logoLink to Journal of Virology
. 1979 Jul;31(1):190–198. doi: 10.1128/jvi.31.1.190-198.1979

Genetic Analysis of a Baculovirus, Autographa californica Nuclear Polyhedrosis Virus I. Isolation of Temperature-Sensitive Mutants and Assortment into Complementation Groups

Martha Brown 1, Allan M Crawford 1, Peter Faulkner 1
PMCID: PMC353435  PMID: 16789180

Abstract

Temperature-sensitive (ts) mutants were isolated from the baculovirus Autographa californica (alfalfa looper) MNPV, grown in Spodoptera frugiperda (fall armyworm) cells in the presence of N-methyl-N′-nitro-N-nitrosoguanidine. Of 567 plaque isolates screened, 27 were temperature sensitive (ts), representing a mutation frequency of 4.8%. Ten ts mutants were studied in detail: six failed to yield nonoccluded virus at 33°C (NOV mutants), whereas the other four produced nonoccluded virus but were restricted in formation of polyhedra at 33°C (Poly mutants). One of the six NOV mutants failed to synthesize viral DNA. Reversion and leak frequencies were determined, and the mutants were assorted into complementation groups based on the yield of polyhedrin synthesis in cells coinfected with pairs of mutants at 33°C, as measured by radioimmunoassay. For NOV mutants, complementation indexes were also based on virus yield and were consistent with those based on polyhedrin synthesis. Nine mutants were assorted into five complementation groups. One mutant remained unclassified.

Full text

PDF
190

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Brown D. A., Bud H. M., Kelly D. C. Biophysical properties of the structural components of a granulosis virus isolated from the cabbage white butterfly (Pieris brassicae). Virology. 1977 Sep;81(2):317–327. doi: 10.1016/0042-6822(77)90148-9. [DOI] [PubMed] [Google Scholar]
  2. Brown M., Faulkner P. Plaque assay of nuclear polyhedrosis viruses in cell culture. Appl Environ Microbiol. 1978 Jul;36(1):31–35. doi: 10.1128/aem.36.1.31-35.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bud H. M., Kelly D. C. The DNA contained by nuclear polyhedrosis viruses isolated from four Spodoptera spp. (Lepidoptera, Noctuidae); genome size and configuration assessed by electron microscopy. J Gen Virol. 1977 Oct;37(1):135–143. doi: 10.1099/0022-1317-37-1-135. [DOI] [PubMed] [Google Scholar]
  4. CAIRNS J. The initiation of vaccinia infection. Virology. 1960 Jul;11:603–623. doi: 10.1016/0042-6822(60)90103-3. [DOI] [PubMed] [Google Scholar]
  5. Crawford A. M., Faulkner P., Kalmakoff J. Comparison of solid-phase radioimmunoassays for baculoviruses. Appl Environ Microbiol. 1978 Jul;36(1):18–24. doi: 10.1128/aem.36.1.18-24.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Esparza J., Purifoy D. J., Schaffer P. A., Benyesh-Melnick M. Isolation, complementation and preliminary phenotypic characterization of temperature-sensitive mutants of herpes simplex virus type 2. Virology. 1974 Feb;57(2):554–565. doi: 10.1016/0042-6822(74)90194-9. [DOI] [PubMed] [Google Scholar]
  7. Harrap K. A., Payne C. C., Robertson J. S. The properties of three baculoviruses from closely related hosts. Virology. 1977 Jun 1;79(1):14–31. doi: 10.1016/0042-6822(77)90330-0. [DOI] [PubMed] [Google Scholar]
  8. Jamieson A. T., Subak-Sharpe J. H. Interallelic complementation of mutants of herpes simplex virus deficient in deoxypyrimidine kinase activity. Virology. 1978 Mar;85(1):109–117. doi: 10.1016/0042-6822(78)90415-4. [DOI] [PubMed] [Google Scholar]
  9. Kelly D. C. The DNA contained by nuclear polyhedrosis viruses isolated from four Spodoptera sp. (Lepidoptera, Noctuidae): genome size and homology assessed by DNA reassociation kinetics. Virology. 1977 Jan;76(1):468–471. doi: 10.1016/0042-6822(77)90325-7. [DOI] [PubMed] [Google Scholar]
  10. Knudson D. L., Harrap K. A. Replication of nuclear polyhedrosis virus in a continuous cell culture of Spodoptera frugiperda: microscopy study of the sequence of events of the virus infection. J Virol. 1975 Jan;17(1):254–268. doi: 10.1128/jvi.17.1.254-268.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Knudson D. L., Tinsley T. W. Replication of a nuclear polyhedrosis virus in a continuous cell culture of Spodoptera frugiperda: purification, assay of infectivity, and growth characteristics of the virus. J Virol. 1974 Oct;14(4):934–944. doi: 10.1128/jvi.14.4.934-944.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Knudson D. L., Tinsley T. W. Replication of a nuclear polyhedrosis virus in a continuous cell line of Spodoptera frugiperda: partial characterization of the viral DNA, comparative DNA-DNA hybridization, and patterns of DNA synthesis. Virology. 1978 Jun 1;87(1):42–57. doi: 10.1016/0042-6822(78)90156-3. [DOI] [PubMed] [Google Scholar]
  13. MacKinnon E. A., Henderson J. F., Stoltz D. B., Faulkner P. Morphogenesis of nuclear polyhedrosis virus under conditions of prolonged passage in vitro. J Ultrastruct Res. 1974 Dec;49(3):419–435. doi: 10.1016/s0022-5320(74)90055-0. [DOI] [PubMed] [Google Scholar]
  14. Miller L. K., Dawes K. P. Restriction endonuclease analysis for the identification of baculovirus pesticides. Appl Environ Microbiol. 1978 Feb;35(2):411–421. doi: 10.1128/aem.35.2.411-421.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Potter K. N., Faulkner P., MacKinnon E. A. Strain selection during serial passage of Trichoplusia in nuclear polyhedrosis virus. J Virol. 1976 Jun;18(3):1040–1050. doi: 10.1128/jvi.18.3.1040-1050.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Schaffer P. A., Aron G. M., Biswal N., Benyesh-Melnick M. Temperature-sensitive mutants of herpes simplex virus type 1: isolation, complementation and partial characterization. Virology. 1973 Mar;52(1):57–71. doi: 10.1016/0042-6822(73)90398-x. [DOI] [PubMed] [Google Scholar]
  17. Shvedchikova N. G., Ulanov B. P., Tarasevich L. M. Elektronnomikroskopicheskoe issledovanie dnk, vydelennoi iz virusnykh chastits pri granuleze sibirskogo shelkopriada. Vopr Virusol. 1968 Sep-Oct;13(5):631–632. [PubMed] [Google Scholar]
  18. Summers M. D., Anderson D. L. Characterization of nuclear polyhedrosis virus DNAs. J Virol. 1973 Dec;12(6):1336–1346. doi: 10.1128/jvi.12.6.1336-1346.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Summers M. D., Anderson D. L. Granulosis virus deoxyribonucleic acid: a closed, double-stranded molecule. J Virol. 1972 Apr;9(4):710–713. doi: 10.1128/jvi.9.4.710-713.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Summers M. D., Egawa K. Physical and chemical properties of Trichoplusia ni granulosis virus granulin. J Virol. 1973 Nov;12(5):1092–1103. doi: 10.1128/jvi.12.5.1092-1103.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Summers M. D., Volkman L. E., Hsieh C. Immunoperoxidase detection of baculovirus antigens in insect cells. J Gen Virol. 1978 Sep;40(3):545–557. doi: 10.1099/0022-1317-40-3-545. [DOI] [PubMed] [Google Scholar]
  22. Tinsley T. W. The potential of insect pathogenic viruses as pesticidal agents. Annu Rev Entomol. 1979;24:63–87. doi: 10.1146/annurev.en.24.010179.000431. [DOI] [PubMed] [Google Scholar]
  23. Vaughn J. L., Goodwin R. H., Tompkins G. J., McCawley P. The establishment of two cell lines from the insect Spodoptera frugiperda (Lepidoptera; Noctuidae). In Vitro. 1977 Apr;13(4):213–217. doi: 10.1007/BF02615077. [DOI] [PubMed] [Google Scholar]
  24. Volkman L. E., Summers M. D., Hsieh C. H. Occluded and nonoccluded nuclear polyhedrosis virus grown in Trichoplusia ni: comparative neutralization comparative infectivity, and in vitro growth studies. J Virol. 1976 Sep;19(3):820–832. doi: 10.1128/jvi.19.3.820-832.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES