Skip to main content
PMC home page
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1987 Jul;53(7):1412–1417. doi: 10.1128/aem.53.7.1412-1417.1987

Baculovirus interaction with nontarget organisms: a virus-borne reporter gene is not expressed in two mammalian cell lines.

L F Carbonell 1, L K Miller 1
PMCID: PMC203885  PMID: 3116925

Abstract

The safety of baculoviruses with respect to mammalian species was studied by using a genetically engineered recombinant of Autographa californica nuclear polyhedrosis virus. This recombinant contains the chloramphenicol acetyltransferase (CAT) gene under the control of a mammalian-active promoter and expresses substantial levels of CAT activity on infection of permissive and nonpermissive insect cells (L.F. Carbonell, M.J. Klowden, and L.K. Miller, J. Virol. 56:153-160, 1985). Extremely low levels of CAT activity were detected in mouse and human cell lines that were continuously exposed to the A. californica nuclear polyhedrosis virus recombinant. The appearance of CAT was not inhibited by cycloheximide. Isopycnic centrifugation of purified inoculum showed that a low level of CAT activity was associated with the insect-derived viral particles. Thus, the observed CAT activity is carried into the cells with the virus inoculum, and active expression of the baculovirus-borne CAT gene is not observed in either cell line. The inability of the CAT gene to be expressed in these cell lines with this model system provides additional assurance of the safety of insect baculoviruses with respect to mammalian species.

Full text

PDF

Images in this article

1414Image
on p.1414
1415Image
on p.1415

Selected References

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

  1. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  2. Carbonell L. F., Klowden M. J., Miller L. K. Baculovirus-mediated expression of bacterial genes in dipteran and mammalian cells. J Virol. 1985 Oct;56(1):153–160. doi: 10.1128/jvi.56.1.153-160.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Gorman C. M., Merlino G. T., Willingham M. C., Pastan I., Howard B. H. The Rous sarcoma virus long terminal repeat is a strong promoter when introduced into a variety of eukaryotic cells by DNA-mediated transfection. Proc Natl Acad Sci U S A. 1982 Nov;79(22):6777–6781. doi: 10.1073/pnas.79.22.6777. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Gorman C. M., Moffat L. F., Howard B. H. Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol Cell Biol. 1982 Sep;2(9):1044–1051. doi: 10.1128/mcb.2.9.1044. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Gröner A., Granados R. R., Burand J. P. Interaction of Autographa californica nuclear polyhedrosis virus with two nonpermissive cell lines. Intervirology. 1984;21(4):203–209. doi: 10.1159/000149522. [DOI] [PubMed] [Google Scholar]
  6. Ignoffo C. M. Effects of entomopathogens on vertebrates. Ann N Y Acad Sci. 1973 Jun 22;217:141–172. doi: 10.1111/j.1749-6632.1973.tb32756.x. [DOI] [PubMed] [Google Scholar]
  7. Lee H. H., Miller L. K. Isolation of genotypic variants of Autographa californica nuclear polyhedrosis virus. J Virol. 1978 Sep;27(3):754–767. doi: 10.1128/jvi.27.3.754-767.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Maeda S., Kawai T., Obinata M., Fujiwara H., Horiuchi T., Saeki Y., Sato Y., Furusawa M. Production of human alpha-interferon in silkworm using a baculovirus vector. Nature. 1985 Jun 13;315(6020):592–594. doi: 10.1038/315592a0. [DOI] [PubMed] [Google Scholar]
  9. Miller L. K., Lingg A. J., Bulla L. A., Jr Bacterial, viral, and fungal insecticides. Science. 1983 Feb 11;219(4585):715–721. doi: 10.1126/science.219.4585.715. [DOI] [PubMed] [Google Scholar]
  10. Miltenburger H. G., Krieg A. Bioinsecticides: II. Baculoviridae. Adv Biotechnol Processes. 1984;3:291–313. [PubMed] [Google Scholar]
  11. Miyamoto C., Smith G. E., Farrell-Towt J., Chizzonite R., Summers M. D., Ju G. Production of human c-myc protein in insect cells infected with a baculovirus expression vector. Mol Cell Biol. 1985 Oct;5(10):2860–2865. doi: 10.1128/mcb.5.10.2860. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Obrig T. G., Culp W. J., McKeehan W. L., Hardesty B. The mechanism by which cycloheximide and related glutarimide antibiotics inhibit peptide synthesis on reticulocyte ribosomes. J Biol Chem. 1971 Jan 10;246(1):174–181. [PubMed] [Google Scholar]
  13. Pennock G. D., Shoemaker C., Miller L. K. Strong and regulated expression of Escherichia coli beta-galactosidase in insect cells with a baculovirus vector. Mol Cell Biol. 1984 Mar;4(3):399–406. doi: 10.1128/mcb.4.3.399. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Rice W. C., Miller L. K. Baculovirus transcription in the presence of inhibitors and in nonpermissive Drosophila cells. Virus Res. 1986 Nov;6(2):155–172. doi: 10.1016/0168-1702(86)90047-x. [DOI] [PubMed] [Google Scholar]
  15. Schneider I. Cell lines derived from late embryonic stages of Drosophila melanogaster. J Embryol Exp Morphol. 1972 Apr;27(2):353–365. [PubMed] [Google Scholar]
  16. Smith G. E., Ju G., Ericson B. L., Moschera J., Lahm H. W., Chizzonite R., Summers M. D. Modification and secretion of human interleukin 2 produced in insect cells by a baculovirus expression vector. Proc Natl Acad Sci U S A. 1985 Dec;82(24):8404–8408. doi: 10.1073/pnas.82.24.8404. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Tjia S. T., zu Altenschildesche G. M., Doerfler W. Autographa californica nuclear polyhedrosis virus (AcNPV) DNA does not persist in mass cultures of mammalian cells. Virology. 1983 Feb;125(1):107–117. doi: 10.1016/0042-6822(83)90067-3. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Volkman L. E., Goldsmith P. A. In Vitro Survey of Autographa californica Nuclear Polyhedrosis Virus Interaction with Nontarget Vertebrate Host Cells. Appl Environ Microbiol. 1983 Mar;45(3):1085–1093. doi: 10.1128/aem.45.3.1085-1093.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Applied and Environmental Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES