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United States Patent (19) 11 Patent Number: 6,017,734 Summers Et Al

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United States Patent (19) 11 Patent Number: 6,017,734 Summers Et Al USOO6O17734A United States Patent (19) 11 Patent Number: 6,017,734 Summers et al. (45) Date of Patent: Jan. 25, 2000 54 UNIQUE NUCLEOTIDE AND AMINO ACID Ahmad, S., Bassiri, M., Banerjee, AK., Yilma, T. (1993). SEQUENCE AND USES THEREOF Immunological Characterization of the VSV Nucleocapsid (N) Protein Expressed by Recombinant Baculovirus in 75 Inventors: Max D. Summers; Sharon C. SpOdoptera exigua Larva: Use in Differential Diagnosis Braunagel; Tao Hong, all of Bryan, Between Vaccinated and Infected Animals. Virol TeX. ogy. 192:207-216. Arvinte T., Schulz B., Cudd A. Nicolau C. (1989). Low pH 73 Assignee: The Texas A & M University System, asSociation of proteins with the membranes of intact red College Station, Tex. blood cells. I. Exogenous glycophorin and the CD4 mol ecule. Biochim. Biophys. Acta 981:51-60. 21 Appl. No.: 08/792,832 Ayres, M. D., Howard, S. C., Kuzio, J., Lopez-Ferber, M., and Possee, R. D. (1994). The Complete DNA Sequence of 22 Filed: Jan. 30, 1997 Autographa californica Nuclear Polyhedrosis Virus. Virol ogy. 202:586-605. Related U.S. Application Data Belyaev, A. S., and Roy, P. (1993). Development of bacu lovirus triple and quadruple expression vectors: co-expres 63 Continuation-in-part of application No. 08/678,435, Jul. 3, Sion of three or four bluetongue virus proteins and the 1996, abandoned Synthesis of bluetongue virus-like particles in insect cells. 60 Provisional application No. 60/000,955, Jul. 7, 1995. Nucleic Acids Res. 21:1219–1223. 51 Int. Cl." ............................. C07H 21/00; C12N 5/10; Blissard, G. W., and Rohrmann, G. F. (1989). Location, C12N 15/33; C12N 15/63 Sequence, Transcriptional Mapping, and Temporal Expres 52 U.S. Cl. .................... 435/69.7; 435/91.4; 435/320.1; sion of the gp64 Envelope Glycoprotein Gene of the Orgyia 435/348; 435/365; 536/23.1; 536/23.72; pseudotsugata Multicapsid Nuclear Polyhedrosis Virus. 536/24.1 Virology. 170:537–555. Blissard, G.W., and Rohrmann, G. F., (1990). Baculovirus 58 Field of Search .................................. 435/69.1, 69.7, Diversity and Molecular Biology. Annu. Rev. Entomol. 435/69.8, 172.1, 320.1, 325, 348, 365, 35:127-155. 410, 91.4; 514/44; 536/23.1, 23.72, 24.1 Bozzola, J. J., and Russell, L. D. (1992). Electron Micros copy. Jones and Bartlett Publishers, Boston, MA. 56) References Cited Bradford, M. M. (1976). A Rapid and Sensitive Method for the Quantitation of Microgram Quantities of Protein Utiliz FOREIGN PATENT DOCUMENTS ing the Principle of Protein-Dye Binding. Anal. Biochem. O 362526 4/1990 European Pat. Off.. 72:248-254. O SOS 207 A1 3/1992 European Pat. Off.. Braunagel, S. C., et al. (1992). Sequence, Genomic Orga nization of the EcoR1-A Fragment Autographa Californica OTHER PUBLICATIONS Nuclear Polyhedrosis Virus, and Identification of a Viral Encoded Protein Resembling the Outer Capsid Protein VPS Harvey et al. (1983) Biochemical and biological variation of of Rotavirus, Virology. 191:1003-1008. Cydia pomenella (Coding Moth) granulosis virus. (Virology Braunagel, S.C., et al. (1996). Identification and Anaylsis of 124:21-34. an Autographa californica Nuclear Polyhedrosis Virus Structural Protein of the Occlusion-Derived Virus Envelope Kuzio et al. (1984) Regions of repeated DNA in the genome ODV-E35, and ODV-EC27, Virology. 222:(in press). of Choristoneura fumiferana nuclear polyhedrosis virus. Braunagel, S. C., et al. (1996). Transcription, Translation Virology 139:185-188. and Cellular Localization of Three Autographa californica Ma et al. (Jan. 1993) Cloning and sequence analysis of a p40 Nuclear Polyhedrosis Virus Structural Proteins: ODV-E18, Structural protein gene of Helicoverpa Zea nuclear polyhe ODV-E35, and ODV-EC27, Virology. 222:100-114. drosis virus. Virology 192:224-233. Kondo et al. (Jul. 1991) Host range expansion by recombi (List continued on next page.) nation of the baculoviruses Bombyx mori nuclear polyhe Primary Examiner-George C. Elliott drosis virus and Autographa californica nuclear polyhedro ASSistant Examiner Robert Schwartzman sis virus. J. Virol. 65:3625-3632. Attorney, Agent, or Firm Arnold, White & Durkee Leisy et al. (Nov. 1984) Conservation of genome organiza 57 ABSTRACT tion in two multicapsid nuclear polyhedrosis viruses. J. Viro. 52:699-702. Provided are hydrophobic targeting Sequences, which may Serve to target heterologous proteins to a variety of cellular Camacho et al. (1991) Protein p22 of African Swine Fever membranes. In particular, the Structural components of the Virus: an early structural protein that is incorporated into the nuclear envelope, or those components which become membrane of infected cells. Virology 181:251-257. nucleus-associated, may be targeted with the Sequences Friedmann (Jun. 1997) Overcoming the obstacles to gene provided. Also provided are methods of targeting heterolo therapy. Scientific Amer. Jun.:96-101. gous proteins to particular membranes, and the use of these Adams, J.R., and McClintock, J. T.; Baculoviridae. Nuclear targeted proteins in therapeutic, diagnostic and insecticidal Polyhedrosis Virus, In “Atlas of Invertebrate Viruses” (J.R. applications. Adams and J. R. Bonami, Eds.). CRC Press, Boca Raton, FL. (1991) 87–204. 56 Claims, 24 Drawing Sheets 6,017,734 Page 2 OTHER PUBLICATIONS Harel, A., Zlotkin, E., Nainudel-Epszteyn, S., Feinstein, N., Fisher, P. A., and Gruenbaum, Y. (1989). Persistence of Braunagel, S. C., et al. (1994). Autographa californica major nuclear envelope antigens in an envelope-like Struc Nuclear Polyhedrosis Virus, PDV, and ECV Viral Envelopes ture during mitosis in Drosophila melanogaster embryoS.J. and Nucleocapsids: Structural Proteins, Antigens, Lipid and Cell Sci. 94:463-470. Fatty Acid Profiles. Virology. 202:315–328. Harlow, E., and Lane D. (1988). Antibodies: A Laboratory Carrington, J. C., and Freed, D.D. (1990). Cap-Independent Manual. Cold Spring Harbor Laboratory Press, Cold Spring Enhancement of Translation by a Plant Potyvirus 5' Non Harbor, New York, pp. 92-115. translated Region. J. Virol.64:1590–1597. Hershberger, P.A., LaCount, D.J., and Friesen, P. D. (1994). Carson, D. D., Guarino, L. A., and Summers, M. D. (1988). The Apoptotic Suppressor P35 Is Required Early During Functional Mapping of an AcMNPV Immediate Early Gene Baculovirus Replication And Is Targeted to the Cytosol of Which Augments Expression of the IE-1 trans-Activated Infected CellS. J. Virol. 68:3467-3477. 39K Gene. Virology. 162:444–451. Hill, J. E., and Faulkner, P. (1994). Identification of the gp67 gene of a baculovirus pathogenic to the Spruce budworm, Carson, D.D., Summers, M.D., and Guarino, L. A. (1991). Choristoneura fumiferana multinucleocapsid nuclear poly Transient Expression of the Autographa californica Nuclear hedrosis virus. J. Gen. Virol. 75:1811-1813. Polyhedrosis Virus Immediate Early Gene, IE-N, Is Regu Hong, T. (1995). Studies on a PDV Envelope Protein of lated by Three Viral Elements. J. Virol. 65:945–951. Autographa californica Nuclear Polyhedrosis Virus. Ph.D Charlton, C. A., and Volkman, L. E. (1993). Penetration of dissertation, Department of Biology, Texas A&M Univer Autographa californica Nuclear Polyhedrosis Virus Nucleo sity, College Station, Texas. capsids into IPLB Sf21 Cells Induces Actin Cable Forma Hong, T., Braunagel, S. C., and Summers, M. D. (1994). tion. Virology. 197:245-254. Transcription, Translation, and Cellular Localization of Choi, J., and Guarino, L. A. (1995). A Temperature-Sensi PDV-E66: A Structural Protein of the PDV Envelope of tive IE1 Transactivation and DNA Binding Activities. Virol Autographa californica Nuclear Polyhedrosis Virus. Virol ogy. 209:90-98. ogy. 204:210-222. Choi, J., and Guarino, L. A. (1995). Expression of the IE1 Hong, T., Braunagel, S. C., and Summers, M. D. (1996). A Transactivator of Autographa Californica Nuclear Polyhe Baculovirus Envelope Protein Sequence Mediates Protein drosis Virus During Viral Infection. Virology. 209:99-107. Transport to Membranes Within the Cytoplasm, Nuclear Choi, J., and Guarino, L. A. (1995). The Baculoviruses Envelope, Baculovirus-Induced Intranuclear Microvesicles Transactivator IE1 Binds to Viral Enhancer Elements in the and the Envelope of Occlusion Derived Virus. Mole. Cell Absence of Insect Cell Factors. J. Virol. 69:4548–4551. Biol. (Submitted). Hooft van Iddekinge, B. J. L., Smith, G. E., and Summers, Cory, J. S., and Bishop, D. H. L. (1995). Use of Baculovi M. D. (1983). Nucleotide Sequence of the Polyhedrin Gene ruses as Biological Insecticides, In “Methods in Molecular of Autographa califonica Nuclear Polyhedrosis Virus. Virol Biology”. vol.39, pp. 277–294, (C. D. Richardson, ed.). ogy. 131:561-565. Humana Press, Inc., Totowa, N.J. Hunter, T. (1993). Braking the Cycle. Cell. 75:839–841. Devereux, J., Haeberli, P., and Smithies, O. (1984). A Ignoffo, C. M. (1971). Intraperitoneal Injection of White comprehensive Set of Sequence analysis programs for the Mice with Nucleopolyhedrosis Virus of the Beet Army VAX. Nucl. Acids Res. 12:389-395. worm, Spodoptera exigua. J. Invert Pathol. 17:453-454. Dewey, R. E., Levings, C. S., III, and Timothy, D. H. (1986). Ignoffo, C. M., Anderson, R. F., and Woodward, G. (1972). Novel Recombinations in the Maize Mitochondrial Genome Teratogenic Potential in Rats Fed the Nuclear Polyhedrosis Produce a Unique Transcriptional Unit in the Texas Mal Virus of Heliothis. Environmental Entomology. 2:337-338. e-Sterile Cytoplasm. Cell. 44:439-449. Ignoffo, C. M., and Heimpel, A. M. (1965). The Funk, C.J., and Consigli, R. A. (1993). Temporal expression Nuclear-Polyhedrosis Virus of Heliothis zea (Boddie) and and immunogold localization of Plodia interpunctella Heliothis virescens (Fabricius) V. Toxicity-Pathogenicity of granulosis virus structural proteins. Virus Res. 28:57-66. Virus to White Mice and Guinea Pigs. J. Invert. Pathol. Furukawa, K., Pante, N., Aebi, U., and Gerace, L. (1995). 7:329-340. Cloning of a cDNA for lamina associated polypeptide 2 Ignoffo, C. M., and Rafajko, R. R. (1972). In vitro Attempts (LAP2) and identification of regions that specify targeting to to Infect Primate Cells with the Nucleopolyhedrosis Virus of the nuclear envelope.
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