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Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1989 Mar;86(5):1453–1456. doi: 10.1073/pnas.86.5.1453

Complementation of recombinant baculoviruses by coinfection with wild-type virus facilitates production in insect larvae of antigenic proteins of hepatitis B virus and influenza virus.

P M Price 1, C F Reichelderfer 1, B E Johansson 1, E D Kilbourne 1, G Acs 1
PMCID: PMC286714  PMID: 2646635

Abstract

We describe the coinfection of insects with wild-type and recombinant baculoviruses in which the polyhedrin gene promoter is used to express hepatitis B virus envelope protein (hepatitis B virus surface antigen; HBsAg) or influenza A virus neuraminidase (NA). Viruses were administered per os to larvae of the cabbage looper, Trichoplusia ni, causing an infection that within 5 days resulted in the production of approximately 0.15 mg of HBsAg per insect, representing 1.5% of the total extracted protein, or approximately 2.8 mg of NA per insect, representing 28% of the total extractable protein. The HBsAg and NA produced by infected larvae were purified from insect lysates. These proteins were antigenic as determined by conformation-dependent immunoassays. The NA was enzymatically active with conventional substrates. The method of infection described allows genetic complementation by wild-type virus of recombinant viruses lacking the polyhedrin gene essential for infection per os and has implications for the high-yield production in insect larvae of other recombinant proteins of baculoviruses.

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Selected References

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  1. AMINOFF D. Methods for the quantitative estimation of N-acetylneuraminic acid and their application to hydrolysates of sialomucoids. Biochem J. 1961 Nov;81:384–392. doi: 10.1042/bj0810384. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Gallagher M., Bucher D. J., Dourmashkin R., Davis J. F., Rosenn G., Kilbourne E. D. Isolation of immunogenic neuraminidases of human influenza viruses by a combination of genetic and biochemical procedures. J Clin Microbiol. 1984 Jul;20(1):89–93. doi: 10.1128/jcm.20.1.89-93.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Heermann K. H., Goldmann U., Schwartz W., Seyffarth T., Baumgarten H., Gerlich W. H. Large surface proteins of hepatitis B virus containing the pre-s sequence. J Virol. 1984 Nov;52(2):396–402. doi: 10.1128/jvi.52.2.396-402.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Itoh Y., Hayakawa T., Fujisawa Y. Expression of hepatitis B virus surface antigen P31 gene in yeast. Biochem Biophys Res Commun. 1986 Jul 16;138(1):268–274. doi: 10.1016/0006-291x(86)90275-5. [DOI] [PubMed] [Google Scholar]
  5. Khan M. W., Gallagher M., Bucher D., Cerini C. P., Kilbourne E. D. Detection of influenza virus neuraminidase-specific antibodies by an enzyme-linked immunosorbent assay. J Clin Microbiol. 1982 Jul;16(1):115–122. doi: 10.1128/jcm.16.1.115-122.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Kilbourne E. D. Comparative efficacy of neuraminidase-specific and conventional influenza virus vaccines in induction of antibody to neuraminidase in humans. J Infect Dis. 1976 Oct;134(4):384–394. doi: 10.1093/infdis/134.4.384. [DOI] [PubMed] [Google Scholar]
  7. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  8. Lai C. J., Markoff L. J., Zimmerman S., Cohen B., Berndt J. A., Chanock R. M. Cloning DNA sequences from influenza viral RNA segments. Proc Natl Acad Sci U S A. 1980 Jan;77(1):210–214. doi: 10.1073/pnas.77.1.210. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. Myers R. W., Lee R. T., Lee Y. C., Thomas G. H., Reynolds L. W., Uchida Y. The synthesis of 4-methylumbelliferyl alpha-ketoside of N-acetylneuraminic acid and its use in a fluorometric assay for neuraminidase. Anal Biochem. 1980 Jan 1;101(1):166–174. doi: 10.1016/0003-2697(80)90056-1. [DOI] [PubMed] [Google Scholar]
  11. Peluso R. W., Rosenberg G. H. Quantitative electrotransfer of proteins from sodium dodecyl sulfate-polyacrylamide gels onto positively charged nylon membranes. Anal Biochem. 1987 May 1;162(2):389–398. doi: 10.1016/0003-2697(87)90409-x. [DOI] [PubMed] [Google Scholar]
  12. Price P. M., Mohamad A., Zelent A., Neurath A. R., Acs G. Translational selection in the expression of the hepatitis B virus envelope proteins. DNA. 1988 Jul-Aug;7(6):417–422. doi: 10.1089/dna.1.1988.7.417. [DOI] [PubMed] [Google Scholar]
  13. Smith G. E., Fraser M. J., Summers M. D. Molecular Engineering of the Autographa californica Nuclear Polyhedrosis Virus Genome: Deletion Mutations Within the Polyhedrin Gene. J Virol. 1983 May;46(2):584–593. doi: 10.1128/jvi.46.2.584-593.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Stibbe W., Gerlich W. H. Structural relationships between minor and major proteins of hepatitis B surface antigen. J Virol. 1983 May;46(2):626–628. doi: 10.1128/jvi.46.2.626-628.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Summers M. D., Smith G. E. Baculovirus structural polypeptides. Virology. 1978 Feb;84(2):390–402. doi: 10.1016/0042-6822(78)90257-x. [DOI] [PubMed] [Google Scholar]
  16. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]

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