Bio/Technology
8, 950 - 955 (1990)
doi:10.1038/nbt1090-950
Use of Early Baculovirus Promoters for Continuous Expression and Efficient Processing of Foreign Gene Products in Stably Transformed Lepidopteran CellsDonald L. Jarvis*, 1, 3, Jo-Ann G. W. Fleming1, 3, Gerald R. Kovacs2, 3, Max D. Summers1, 2, 3, 4
& Linda A. Guarino1, 3, 4
1Department of Entomology, College Station, Texas 77843.
2Department of Biochemistry and Biophysics, College Station, Texas 77843.
3Center for Advanced Invertebrate Molecular Sciences, College Station, Texas 77843.
4Texas A&M University, and Texas Agricultural Experiment Station; College Station, Texas 77843.
*Corresponding author. Baculoviruses are currently used as vectors for the transient high-level expression of foreign gene products in insect cells. In this study, we demonstrate that baculoviruses can also be made to continuously express a foreign gene product by using the promoter from IE1, an immediate early viral gene, to produce stably-transformed insect cells. This approach gave levels of foreign gene expression lower than those usually obtained with the lytic baculovirus expression vector system. Expression, however, was continuous and stable, and a complex human glyco-protein (tissue plasminogen activator) was processed more efficiently. We conclude that stable transformation is a feasible approach for baculovirus-mediated foreign gene expression in lepidopteran cells, particularly for products that are relatively poorly-expressed and/or processed in lyrically infected cells.
REFERENCES
- Luckow, V.A. and Summers, M.D. 1988. Trends in the development of baculovirus vectors. Bio/Technology 6: 47−55. | ISI | ChemPort |
- Miller, L.K. 1988. Baculoviruses as gene expression vectors. Ann. Rev. Microbiol. 42: 177−199. | Article | ISI | ChemPort |
- Jarvis, D.L. and Summers, M.D. Baculovirus expression vectors. In: R.E. Isaacson (Ed.) Recombinant DNA Vaccines: Rationale and Strategies. Marcel Dekker, Inc., New York, N.Y. In press.
- Smith, G.E., Fraser, M.J. and Summers, M.D. 1983. Molecular engineering of the Autographa californica nuclear polyhedrosis genome: deletion mutations within the polyhedrin gene. J. Virol. 46: 584−593. | ISI | ChemPort |
- Smith, G.E., Summers, M.D. and Fraser, M.J. 1983. Production of human beta interferon in insect cells infected with a baculovirus expression vector. Mol. Cell. Biol. 3: 2156−2165. | PubMed | ISI | ChemPort |
- Pennock, G.D., Shoemaker, C. and Miller, L.K. 1984. Strong and regulated expression of Escherichia coli
 -galactosidase in insect cells with a baculovirus vector. Mol. Cell. Biol. 4: 399−406. | PubMed | ISI | ChemPort |
- Summers, M.D. and Smith, G.E. 1987. A manual of methods for baculovirus vectors and insect cell culture procedures. Texas Agricultural Experiment Station Bulletin No. 1555.
- Friesen, P.D. and Miller, L.K. 1986. The regulation of baculovirus gene expression. Curr. Top. Microbiol. Immunol. 131: 31−49. | PubMed | ISI | ChemPort |
- Guarino, L.A. 1989. Enhancers of early gene expression, p. 211−219. In: J. Mitsuhashi (Ed.) Invertebrate Cell System Applications, Volume I. CRC Press, Inc., Boca Raton, Florida.
- Jarvis, D.L. and Summers, M.D. 1989. Glycosylation and secretion of human tissue plasminogen activator in recombinant baculovirus-infected insect cells. Mol. Cell. Biol. 9: 214−223. | PubMed | ISI | ChemPort |
- Guarino, L.A. and Summers, M.D. 1986. Functional mapping of a trans-activating gene required for expression of a baculovirus delayed-early gene. J. Virol. 57: 563−571. | PubMed | ISI | ChemPort |
- Guarino, L.A. and Summers, M.D. 1987. Nucleotide sequence and temporal expression of a baculovirus regulatory gene. J. Virol. 61: 2091−2099. | ISI | ChemPort |
- Wigler, M., Perucho, M., Kurtz, D., Dana, S., Pellicer, A., Axel, R. and Silverstein, S. 1980. Transformation of mammalian cells with an amplifiable dominant-acting gene. Proc. Natl. Acad. Sci. U.S.A. 77: 3567−3570. | PubMed | ChemPort |
- Southern, P.J. and Berg, P. 1982. Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region promoter. J. Mol. Appl. Gen. 1: 327−341. | ChemPort |
- Santerre, R.F., Allen, N.E., Hobbs, J.N., Rao, N. and Schmidt, R.J. 1984. Expression of prokaryotic genes for hygromycin B and G418 resistance as dominant-selection markers in mouse L cells. Gene 30: 147−156. | Article | PubMed | ISI | ChemPort |
- Bourouis, M. and Jarry, B. 1983. Vectors containing a prokaryotic dihydrofolate reductase gene transform Drosophila cells to methotrexate resistance. EMBO J. 2: 1099−1104. | PubMed | ISI | ChemPort |
- Rio, D.C. and Rubin, G.M. 1985. Transformation of cultured Drosophila melanogaster cells with a dominant selectable marker. Mol. Cell. Biol. 5: 1833−1838. | PubMed | ISI | ChemPort |
- van der Straten, A., Johansen, H., Sweet, R. and Rosenberg, M. 1989. Efficient expression of foreign genes in cultured Drosophila melanogaster cells using hygromycin B selection, p. 183−195. In: J. Mitsuhashi (Ed.) Invertebrate Cell System Applications, Volume I. CRC Press, Inc., Boca Raton, Florida.
- Maiorella, B., Inlow, D., Shauger, D. and Harano, D. 1988. Large-scale insect cell culture for recombinant protein production. Bio/Technology 6: 1406−1410. | ISI | ChemPort |
- Murhammer, D.W. and Goochee, C.F. 1988. The scaleup of insect cell cultures for recombinant protein production. Bio/Technology 6: 1411−1418. | ISI | ChemPort |
- Miller, D.W. and Miller, L.K. 1982. A virus mutant with an insertion of a copia-like transposable element. Nature 299: 562−564. | PubMed | ISI | ChemPort |
- Fraser, M.J., Smith, G.E. and Summers, M.D. 1983. Acquisition of host cell DNA sequences by baculoviruses: Relationship between host DNA insertions of FP mutants of Autographa californica nuclear polyhedrosis virus. J. Virol. 47: 287−300. | ISI | ChemPort |
- Beames, B. and Summers, M.D. 1988. Comparisons of host cell DNA insertions and altered transcription at the site of insertions in few polyhedra baculovirus mutants. Virology 162: 206−220. | Article | PubMed | ISI | ChemPort |
- Panicali, D., Grzelecki, A. and Huang, C. 1986. Vaccinia virus vectors utilizing the -galactosidase assay for rapid selection of recombinant viruses and measurement of gene expression. Gene 47: 193−199. | Article | PubMed | ISI | ChemPort |
- Luckow, V.A. and Summers, M.D. 1988. Signals important for the high-level expression of foreign genes in Autographa californica nuclear polyhedrosis virus expression vectors. Virology 167: 56−71. | Article | PubMed | ISI | ChemPort |
- Sambrook, J., Fritsch, E.F. and Maniatis, T. 1989. Molecular Cloning: A Laboratory Manual, Second edition. Cold Spring Harbor Press, Cold Spring Harbor, N.Y.
- Vaughn, J.L., Goodwin, R.H., Thompkins, G.J. and McCawley, P. 1977. The establishment of two insect cell lines from the insect Spodoptera frugiperda (Lepidoptera:Noctuidae). In Vitro 13: 213−217. | PubMed | ISI | ChemPort |
- Hirt, B. 1967. Selective extraction of polyoma DNA from infected mouse cell cultures. J. Mol. Biol. 26: 365−369. | PubMed | ISI | ChemPort |
- Southern, E.M. 1975. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J. Mol. Biol. 98: 503−517. | PubMed | ISI | ChemPort |
- Feinberg, A.P. and Vogelstein, B. 1983. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Analyt. Biochem. 132: 6−13. | PubMed | ISI | ChemPort |
- Fleming, J.G.W. and Summers, M.D. 1986. Campoletis sonorensis endoparasitic wasps contain forms of C. sonorensis virus DNA suggestive of integrated and extrachromosomal polydnavirus DNAs. J. Virol. 57: 552−562. | ISI | ChemPort |
- Chirgwin, J.M., Przybyla, A.E., MacDonald, R.J. and Rutter, W.J. 1979. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry 18: 5294−5299. | PubMed | ISI | ChemPort |
- Weaver, R.F. and Weissmann, C. 1979. Mapping of an RNA by a modification of the Berk-Sharp procedure: the 5' termini of 15S -globin mRNA precursor and mature -globin mRNA have identical map coordinates. Nucleic Acids Res. 7: 1175−1193. | PubMed | ISI | ChemPort |
- Kessler, S.W. 1975. Rapid isolation of antigens from cells with a Staphylococcal protein. A-antibody adsorbent: parameters of the interaction of antibody-antigen complexes with protein A. J. Immunol. 115: 1617−1624. | PubMed | ISI | ChemPort |
- Laemmli, U.K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680−685. | PubMed | ISI | ChemPort |
- Towbin, H., Staehelin, T. and Gordon, J. 1979. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc. Natl. Acad. Sci. U.S.A. 76: 4350−4354. | PubMed | ChemPort |
- Blake, M.S., Johnston, K.H., Russell-Jones, G.J. and Gotschlich, E.C. 1984. A rapid, sensitive method for detection of alkaline phosphatase conjugated anti-antibody on western blot. Anal. Biochem. 36: 175−179.
- Harlow, E., Crawford, L.V., Pim, D.C. and Williamson, N.M. 1981. Monoclonal antibodies specific for simian virus 40 tumor antigens. J. Virol. 39: 861−869. | PubMed | ISI | ChemPort |
|
