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Porcine adenovirus type 3 genome |
| RE40930 |
Porcine adenovirus type 3 genome
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| Patent Drawings: | |
| Inventor: |
Reddy, et al. |
| Date Issued: |
October 6, 2009 |
| Application: |
11/518,612 |
| Filed: |
September 8, 2006 |
| Inventors: |
Reddy; Police Seshidhar (Chester Springs, PA)
Tikoo; Suresh (Saskatoon, CA)
Babiuk; Lorne A. (Saskatoon, CA)
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| Assignee: |
Vectogen Pty Ltd (Northy Ryde, AU) |
| Primary Examiner: |
Martinell; James |
| Assistant Examiner: |
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| Attorney Or Agent: |
McAndrews Held & Malloy Ltd. |
| U.S. Class: |
514/44R; 435/320.1; 435/325; 435/455; 435/471; 435/488; 435/69.1 |
| Field Of Search: |
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| International Class: |
A01N 43/04; C12N 15/00; C12N 15/63; C12N 15/74; C12N 15/85; C12N 15/86; C12N 5/00; C12P 21/00; C12P 21/02 |
| U.S Patent Documents: |
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| Foreign Patent Documents: |
0259149; WO 99/08706 |
| Other References: |
Verma et al.; "Gene Therapy-Promises, Problems, and Prospects," Nature, vol. 389, pp. 239-242, Sep. 18, 1997. cited by other.
Eck et al., "Gene-Based Therapy," Goodman & Gilman's the Pharmacological Basis of Therapeutics, 9th Edition, Chapter 5, pp. 77-101, 1996. cited by other.
McCoy et al. Nucleotide and Amino Acid Sequence Analysis of the 100K Protein of a Serotype 3 Porcine Adenovirus. DNA Sequence--The Journal of Sequencing and Mapping, vol. 8, pp. 59-61, 1997. cited by other.
McCoy et al. Porcine Adenovirus 3 hexon gene. GeBank, Accession No. U34592, Jun. 12, 1996. cited by other.
Kleiboeker, S. B. et al. (1993). "Genomic cloning and restriction site mapping of a porcine adenovirus isolate: Demonstration of genomic stability in porcine adenovirus" Arch. Virol. 133(3-4):357-368. cited by other.
Park, J. H. et al. (Jun. 1998). "Sequence analysis of the early region 1B (E1B) of porcine adenvirus type 3" RDA J. Veterinary Sci. 40(1):19-25 (Abstract only). cited by other.
Reddy, P. S. et al. (Nov. 1998). "Sequence and transcription map analysis of early region-1 of porcine adenovirus type-3" Virus Res. 58(1-2):97-106. cited by other.
Reddy, P. S. et al. (Nov. 25, 1998). "Nucleotide sequence and transcription map or porcine adenovirus type-3" Virol. 251(2):414-426. cited by other.
Reddy, P. S. et al. (Mar. 1999). "Development of porcine adenovirus-3 as an expression vector" J. Gen. Virol. 80(3):563-570. cited by other.
Animal Cell Culture Ed. R. Freshney, Oxford: IRL Press, 1986. Title page and table of contents only. cited by other.
Antibodies: A Laboratory Manual, Eds. Harlow & Lane. New York: Cold Spring Harbor Press, 1988. Title page and table of contents only. cited by other.
Ball et al. (1988). "Identification of Mouse Adenovirus Type 1 Early Region 1: DNA Sequence and a Conserved Transactivating Function," Journal of Virology, vol. 62, No. 11, 3947-3957. cited by other.
Berk, Arnold and Sharp, Phillip. (1977). "Sizing and Mapping of Early Adenovirus mRNAs by Gel Electroporesis of S1 Endonuclease-Digested Hybrids," Cell vol. 12, 721-732. cited by other.
Brennan, Sean and Savage, Robert. (1990). "Embryonic transcriptional activation of a Xenopus cytopus actine gene does not require a serum response element," Roux's Arch Dev. Biol vol. 199, 89-96. cited by other.
Chartier, C. et al. (1996). "Efficient Generation of Recombinant Adenovirus Vectors by Homologous Recombination in Escherichia coli," Journal of Virology vol. 70, No. 7, 4805-4810. cited by other.
Chiocca, S., et al. (1996). "The Complete DNA Sequence and Genomic Organization of the Avian Adenovirus CELO" Journal of Virology vol. 70, No. 5, 2939-2949. cited by other.
Current Protocols in Molecular Biology, Eds. Ausubel et al. New York: John Wiley & Sons, 1995. Title page and table of contents only. cited by other.
Derbyshire, et al. (1975). "Serological and Pathogenicity Studies with Some Unclassified Porcine Adenoviruses," J. Comp. Path. vol. 85, 437-443. cited by other.
Derbyshire, J.B. "Adenovirus" Disesess of Swine, Ed. Leman, et al 7th ed. Ames. IA: Iowa State University Press, pp. 225-227, 1992. cited by other.
DNA Cloning: A Practical Approach, Ed. D. Glover. Oxford: IRL Press, vols. I, II & III, 1985, 1987. Title page and table of contents only. cited by other.
Fallaux, et al. (1996). "Characterization of 911: A New Helper Cell Line for the Titration and Propagation of Early Region 1-Deleted Adenoviral Vectors," Human Gene Therapy vol. 7, 215-222. cited by other.
Fallaux, et al. (1998). "New Helper Cells and Matched Early Region 1-Deleted Adenovirus Vectors Prevent Generation of Replication-Competent Adenoviruses," Human Gene Therapy vol. 9, 1909-1917. cited by other.
Gerard, Robert and Meidell, Robert. (1993). "Adenovirus-Mediated Gene Transfer," TCM vol. 3, No. 5, 171-177. cited by other.
Gorman, et al. (1982). "Recombinant Genomes Which Express Chloramphenicol Acetyltransferase in Mammalian Cells," Molecular and Cellular Biology vol. 2, No. 9, 1044-1051. cited by other.
Graham, et al. (1977). "Characteristics of a Human Cell Line Transformed by DNA from Human Adenovirus Type 5," J. General Virology vol. 36, 59-72. cited by other.
Graham, et al. (1991). "Gene transfer and expression protocols," Methods in Molecular Biology vol. 7, 109-128. cited by other.
Grunhaus, A. and Horwitz, M.S. (1992). "Adenoviruses as cloning vectors," seminars in Virology vol. 3, 237-252. cited by other.
Hanahan, Douglas (1983). "Studies on Transformation of Escherichia coliwith Plasmids," J. Mol. Biol. vol. 166, 557-580. cited by other.
Hehir, et al. (1996). "Molecular Characterization of Replication-Competent Variants of Adenovirus Vectors and Genome Modifications To Prevent Their Occurrence," Journal of Virology vol. 70, No. 12, 8459-8467. cited by other.
Hirahara et al. (1989). "Isolation of Porcine Adenovirus from the Respiratory Tract of Pigs in Japan," Jpn. J. Vet. Sci. vol. 52, No. 2, 407-409. cited by other.
Hirt (1967). "Selective Extraction of Polyoma DNA from Infected Mouse Cell Cultures," J. Mol. Biol. vol. 26, 365-369. cited by other.
Imler, et al. (1995) "Novel complementation cells lines derived from human lung carcinoma A549 cells support the growth of E1-deleted adenovirus vectors." Gene Therap. vol. 3. 75-84. cited by other.
Imler, et al. (1995) "Trans-Complementation of E1-Deleted Adenovirus: A New Vector to Reduce the Possibility of Codissemination of Wild-Type and Recombinant Adenoviruses." Human Gene Therapy vol. 6, 711-721. cited by other.
Kleiboeker, S. "Sequence analysis of putative E3, pVIII, and fiber genomic regions of a porcine adenovirus," Virus Research vol. 36, 97-106 (1995). cited by other.
Kleiboeker, S. (1995). "Identification and sequence analysis of the E1 genomic region of a porcine adenovirus," Virus Research vol. 36, 259-268. cited by other.
Kleiboeker, S. (1995). "Sequence analysis of the fiber genomic region of a porcine adenovirus predicts a novel fiber protein," Virus Research vol. 39, 299-309. cited by other.
Klonjowski, et al. (1997). "A Recombinant E1-Deleted Canine Adenoviral Vector Capable of Transduction and Expression of a Transgene in Human-Derived Cells and In Vivo." Human Gene Therapy vol. 8. 2103-2115. cited by other.
Kunkel, et al. (1987). "Rapid and Efficient Site-Specific Mutagenesis without Phenotypic Selection," Methods in Enzymology vol. 154, 367-382. cited by other.
Ma, Yuliang and Mathews, Michael B. (1996). "Structure, Function, and Evolution of Adenovirus-Associated RNA: a Phylogenetic Approach," Journal of Virology vol. 70, No. 8, 5083-5099. cited by other.
Maniatis, et al., Molecular Cloning: A Laboratory Manual, New York: Cold Spring Harbor Press, 1982. Title page and table of contents only. cited by other.
McCoy, et al. (1996). "Genomic location and nucleotide sequence of a porcine adenovirus penton base gene," Archives of Virology vol. 141, 1367-1375. cited by other.
McCoy, et al. (1996). "Nucleotide and amino acid sequence analysis of the procine adenovirus 23K protein," DNA Sequence--The Journal of Sequencing and Mapping vol. 6, 251-254. cited by other.
Methods in Nonradioactive Detection, Ed. Howard. Norwalk: Appleton & Lange, 1993. Title page and table of contents only. cited by other.
Morrison, et al. (1997). "Complete DNA sequence of canine adenovirus type 1," Journal of General Virology vol. 78. 873-878. cited by other.
Nonistopic DNA Probe Techniques, Ed. Kricka, San Diego: Academic Press, 1992. Title page and table of contents only. cited by other.
Nonradioactive Labeling and Detection of Biomolecules, Ed. Kessler., Berlin: Springer-Verlag, 1992. Title page and table of contents only. cited by other.
Nucleic Acid Hybridisation: A Practical Approach, Eds. B.D. Harnes and S.J. Higgins. Oxford: IRL Press Ltd., 1985. Title page and table of contents only. cited by other.
Oligonycleotide Synthesis, Ed. M.J., Gait, Oxford: IRL Press, 1984. Title page or table of contents only. cited by other.
Perbal, Bernard. A Practical Guide to Molecular Cloning, New York: John Wiley & Sons, 1984. Title page and table of contents only. cited by other.
Reddy, et al. (1995), "Comparison of the Inverted Repetition Sequences from Five Porcine Adenovirus Serotypes," Virology vol. 212, 237-239. cited by other.
Reddy, et al. (1995), "Molecular cloning and physical mapping of porcine adenovirus types 1 and 2," Archives of Virology vol. 140, 195-200. cited by other.
Reddy, et al. (1995). "Sequence analysis of putative pVIII, E3 and fibre regions of porcine adenovirus type 3," Virus Research vol. 36, 97-106. cited by other.
Reddy, et al. (1996). "Porcine adenoviruses type 1, 2 and 3 have short and simple early E-3 regions," Virus Research vol. 43, 99-109. cited by other.
Reddy, et al. (1997). "Characterization of the Early Region 4 of Porcine Adenovirus Type 3," Virus Genes vol. 15, No. 1, 87-90. cited by other.
Reddy, et al. (1998). "Nucleotide Sequence, Genome Organization, and Transcription Map of Bovine Adenovirus Type 3," Journal of Virology vol. 72, No. 2, 1394-1402. cited by other.
Reddy, et al. (1993). "Restriction Endonuclease Analysis and Molecular Cloning of Porcine Adenovirus Type 3," Intervirology vol. 36, 161-168. cited by other.
Sambrook, et al., Molecular Cloning: A Laboratory Manual, 2.sup.nd ed. New York: Cold Spring Harbor Laboratory Press, Buples 1, 2, & 3 1989. Title page and table of contents only. cited by other.
Shaw, W.V. (1975). "Chloramphenicol Acetyltransferase from Chloramphenicol-Resistant Bacteria" Meth, in Enzymology vol. 43, 737-755. cited by other.
Transcription and Translation, Eds. B. Hames and S. Higgins. Oxford: IRL Press, 1984, Title page and tasble of contents only. cited by other.
Tuboly, et al. (1993). "Potential viral vectors for the stimulation of mucosal antibody responses against enteric viral antigens in pigs," Research in Veterinary Science vol. 54, 345-350. cited by other.
Viral Diarrheas of Man and Animals, Eds. Saif et al., "Enteric Virus Vaccines: Theoretical Considerations, Current Status, and Future Approaches," Chapter 14, pp. 313-329, 1990. cited by other.
Vrati, et al. (1996). "Unique Genome Arrangement of an Ovine Adenovirus: Identification of New Proteins and Proteinase Cleavage Sites," Virology vol. 220, 186-199. cited by other.
Xiang, et al. (1996). "A Replication-Defective Human Adenovirus Recombinant Serves as a Highly Efficacious Vaccine Carrier," Virology vol. 219, 220-227. cited by other.
Zheng, et al. (1994). "The E1 sequence of bovine adenovirus type 3 and complementation of human adenovirus type 5 E1A function in bovine cells," Virus Research vol. 31, 163-186. cited by other.
Zoller, Mark and Smith, Michael. (1982). "Oligonucleotide-directed mutagensis using M13-derived vectors: and efficient and general procedure for the production of point mutations in any fragment of DNA," Nucleac Acids Research vol. 10, No. 20,6487-6500. cited by other.
McCoy et al. Nucleotide and amino acid sequence analysis of the 100k protien of a serotype 3 porcine adenovirus. GeBank, Accession No. U82628, Dec. 18, 1996. cited by other.
Redeclaration, jan. 24, 2006. cited by other.
Judgment, Aug. 9, 2006, patent Interference No. 105358. cited by other.
Board Order Relating to Motion Times. cited by other.
Johnson List of Intended Preliminary Motions, Paper 18. cited by other.
Bett et al.; "An efficient and flexible system for construction of adenovirus vectors eith insertions or deletions in early regions 1 and 3," Proc. Natl. Acad. Sci. USA, vol. 91, pp. 8802-8806, Sep. 1994. cited by other.
Crouzel et al.; "Recombinational construction o fEscherichia coli of infections adenoviral genomes," Proc. Natl. Acad. Sci. USA, vol. 94, pp. 1414-1419, Feb. 1997. cited by other.
Ketner et al.; Efficient manipulation of the human adenovirus genome as an infectious yeast artificial chromosomes clone, Proc. Natl. Acad. Sci. USA, vol. 91, pp. 6186-6190, Jun. 1994. cited by other.
He et al.; "A simplified system for generating recombinant adenoviruses," Proc. Natl. Acad. Sci. USA,vol. 95, pp. 2509-2514, Mar. 1998. cited by other. |
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| Abstract: |
The complete nucleotide sequence of the genome of porcine adenovirus type 3 (PAV-3) is provided. Methods for construction of infectious PAV genomes by homologous recombination in procaryotic cells are provided. Recombinant PAV viruses are obtained by transfection of mammalian cells with recombinant PAV genomes. The PAV-3 genome can be used as a vector for the expression of heterologous nucleotide sequences, for example, for the preparation and administration of subunit vaccines to swine or other mammals. In addition, PAV-3 vectors can be used for gene therapy and expression of heterologous polypeptides. PAV-3 genome sequences can also be used for diagnostic purposes, to detect the presence of PAV-3 DNA in a subject or biological sample. |
| Claim: |
What is claimed is:
1. A replication-defective recombinant PAV-3 vector, wherein said PAV-3 vector is capable of duplex formation under conditions of high stringency with the genome of PAV-3 asdepicted in SEQ ID NO:1, or its complement, said vector comprising ITR sequences, packaging sequences, and at least one heterologous nucleotide sequence, and wherein the PAV-3 vector lacks E1 function.
2. The replication-defective recombinant PAV-3 vector according to claim 1, wherein the vector is deleted in the E1 region.
3. The replication-defective recombinant PAV-3 vector according to claim 2, wherein the vector is additionally deleted in a region selected from the group consisting of E2, E3, E4, L1, L2, L3, L4, L5, L6 and the region between E4 and the rightthe genome.
4. The replication-defective recombinant PAV-3 vector according to claim 3, wherein the vector is additionally deleted in more than one region selected from the group consisting of E2, E3, E4, L1, L2, L3, L4, L5, L6 and the region between E4and the end of the genome.
5. The replication-defective recombinant PAV-3 vector according to claim 1, wherein the heterologous nucleotide sequence encodes a polypeptide selected from the group consisting of coagulation factors, growth hormones, cytokines, lymphokines,tumor-suppressing polypeptides, cell receptors, ligands for cell receptors, protease inhibitors, antibodies, toxins, immunotoxins, dystrophins, cystic fibrosis transmembrane conductance regulator (CFTR) and immunogenic polypeptides.
6. A method for producing a recombinant PAV-3 that comprises introducing the PAV-3 vector of claim 1 into a helper cell line comprising E1 function and recovering virus from the infected cells.
7. A host cell comprising the vector of claim 1.
8. A method for producing a recombinant polypeptide, the method comprising: (a) providing a population of host cells according to claim 7, and (b) growing said population of cells under conditions whereby the polypeptide is expressed.
9. A composition comprising the replication-defective recombinant PAV-3 vector of claim 1.
10. The composition according to claim 9 further comprising a pharmaceutically acceptable vehicle.
11. A composition capable of inducing an immune response in a mammalian subject, said composition comprising a replication-defective recombinant PAV-3 vector according to claim 1, wherein the vector comprises a heterologous nucleotide sequencethat encodes an immunogenic polypeptide; and a pharmaceutically acceptable vehicle.
12. The composition according to claim 11, wherein said immunogenic polypeptide is a pathogen antigen.
13. A recombinant PAV-3 vector comprising a PAV-3 genome capable of duplex formation under conditions of high stringency to the PAV-3 genome as depicted in SEQ ID NO:1, or a complement thereof and at least one heterologous nucleotide sequence,wherein the heterologous nucleotide sequence is inserted in a region selected from the group consisting of E1 region, .[.the E3 region,.]. .Iadd.and .Iaddend.the E4 region .[.and the region between E4 and the right end.]. of the genome.
14. The recombinant PAV-3 vector of claim 13, comprising two or more heterologous nucleotide sequences.
15. The recombinant PAV-3 vector of claim 14, wherein the two or more heterologous nucleotide sequences are inserted at different insertion sites.
16. The recombinant PAV-3 vector of claim 13, wherein the heterologous nucleotide sequence encodes a polypeptide selected from the group consisting of coagulation factors, growth hormones, cytokines, lymphokines, tumor-suppressing polypeptides,cell receptors, ligands for cell receptors, protease inhibitors, antibodies, toxins, immunotoxins, dystrophins, cystic fibrosis transmembrane conductance regulator (CFTR) and immunogenic polypeptides.
17. A host cell comprising the vector of claim 13.
18. A method for producing a recombinant polypeptide, the method comprising: (a) providing a population of host cells according to claim 17, and (b) growing said population of cells under conditions whereby the polypeptide is expressed.
19. A composition comprising the vector of claim 13.
20. The recombinant PAV-3 vector according to claim 13 wherein the heterologous nucleotide sequence is inserted in the E1 region.
.[.21. The recombinant PAV-3 vector according to claim 13 wherein the heterologous nucleotide sequence is inserted in the E3 region..].
22. The recombinant PAV-3 vector according to claim 13 wherein the heterologous nucleotide sequence encodes an immunogenic polypeptide.
23. The recombinant PAV-3 vector according to claim 22, wherein said immunogenic polypeptide is a pathogen antigen.
24. The recombinant PAV-3 vector of claim 13, wherein said vector is replication competent.
25. A method for obtaining a recombinant PAV-3 comprising a heterologous nucleotide sequence inserted into a PAV-3 insertion site .Iadd.located in a region selected from the group consisting of the E1 region, and the E4 region of the PAV-3genome.Iaddend., the method comprising the steps of: (a) providing a PAV-3 genome capable of duplex formation under conditions of high stringency to the PAV-3 genome as depicted in SEQ ID NO: 1; (b) providing a heterologous nucleotide sequence; (c)linking the heterologous nucleotide sequence to guide sequences, the guide sequences being capable of duplex formation under conditions of high stringency to said PAV-3 genome sequences flanking the PAV-3 insertion site, or the complement of saidsequences, such that guide sequences are present at both ends of the heterologous sequence; (d) introducing the construct from step (c) into a cell together with the PAV-3 genome; (e) allowing homologous recombination to occur between the two sequencesfrom step (d) to generate a recombinant PAV-3 genome; (f) purifying the recombinant PAV-3 genome; (g) inserting the recombinant PAV-3 genome into a mammalian cell; (h) culturing the mammalian cell under conditions wherein the recombinant PAV-3 genomeis replicated and packaged; and (i) optionally collecting the recombinant PAV-3 from the cell or the culture medium.
.[.26. The method according to claim 25 wherein the insertion site is located in a region of the PAV-3 genome selected from the group consisting of the E1 region, the E3 region, the E4 region and the region between E4 and the right end of thegenome..].
27. The method according to .[.claim 26.]. .Iadd.claim 25 .Iaddend.wherein the PAV-3 genome is deleted in a region selected from the group consisting of the E1 region, the E3 region, the E4 region and the region between E4 and the right end ofthe genome.
28. A composition comprising a recombinant PAV-3 obtained according to the method of claim 25.
29. The method according to claim 25 wherein said insertion site is E1.
.[.30. The method according to claim 25 wherein said insertion site is E3..].
31. A method for eliciting an immune response in a mammalian host comprising administering a composition comprising a recombinant PAV-3 vector that .[.expresses an immunogenic polypeptide.]. .Iadd.comprises a heterologous nucleic acid thatencodes an immunogenic polypeptide inserted at an insertion site selected from the group consisting of the E1 region, and the E4 region of the PAV-3 genome.Iaddend., wherein said PAV-3 vector is capable of duplex formation under conditions of highstringency to the PAV-3 genome as depicted in SEQ ID NO:1, or a complement thereof; and a pharmaceutically acceptable vehicle.
32. The method of claim 31 wherein said PAV-3 vector lacks E1 function.
33. The method of claim 32 wherein said PAV-3 vector is additionally deleted in a region selected from the group consisting of E2, E3, E4, L1, L2, L3, L4, L5, L6 and the region between E4 and the right end of the genome.
34. The method according to claim 31 wherein said immunogenic polypeptide is a pathogen antigen.
.[.35. A method for obtaining a full-length genomic clone of a PAV-3 genome, the method comprising: (a) providing two or more cloned segments of the PAV-3 genome, wherein said segments are capable of duplex formation under conditions of highstringency to the PAV-3 genome as depicted in SEQ ID NO:1, or a complement thereof, and wherein the cloned segments of the PAV-3 genome, taken together, represent the entire PAV-3 genome; (b) introducing the two or more cloned segments of the PAV-3genome into a cell; (c) allowing homologous recombination to occur within the cell between the two or more cloned segments of the PAV-3 genome to generate a full-length PAV-3 genome; and (d) optionally purifying the full-length PAV-3 genome from thecell..].
.[.36. The method of claim 35 wherein said cell is procaryotic cell..].
37. The method of claim 36 wherein bacterial cell is E. coli.
38. A method for obtaining a recombinant PAV-3 comprising a heterologous nucleotide sequence inserted into a PAV-3 insertion site .Iadd.located in a region selected from the group consisting of the E1 region and the E4 region of the PAV-3genome.Iaddend., comprising the steps of a) introducing a recombinant plasmid into a host cell in combination with a PAV-3 genome, wherein said plasmid comprises a heterologous nucleotide sequence flanked by nucleotide sequences that are capable ofduplex formation under conditions of high stringency to PAV-3 nucleotide sequences flanking the PAV-3 insertion site, or the complement of said sequences, and wherein the PAV-3 genome is capable of duplex formation under conditions of high stringency tothe PAV-3 genome as depicted in SEQ ID NO:1, or a complement thereof, b) allowing homologous recombination to occur between the plasmid and the PAV-3 genome thereby generating a recombinant PAV-3 comprising said heterologous nucleotide sequences; c)isolating said recombinant PAV-3; d) introducing said isolated PAV-3 into a mammalian cell permissive for growth of said PAV-3; e) culturing said mammalian cell under conditions suitable for PAV-3 replication and packaging; and f) optionally,collecting said recombinant PAV-3 produced from step e).
39. The method of claim 38 wherein said PAV-3 vector lacks E1 function.
40. The method of claim 39 wherein said PAV-3 vector is additionally deleted in a region selected from the group consisting of E2, E3, E4, L1, L2, L3, L4, L5, L6 and the region between E4 and the right end of the genome.
41. The replication-defective recombinant PAV-3 vector according to claim 1 wherein the heterologous nucleotide sequence encodes an immunogenic polypeptide.
42. The replication-defective PAV-3 vector according to claim 41, wherein said immunogenic polypeptide is a pathogen antigen.
43. A vaccine for protecting a mammalian host against infection comprising a recombinant PAV-3 vector comprising a PAV-3 genome capable of duplex formation under conditions of high stringency to the PAV-3 genome as depicted in SEQ ID NO:1, or acomplement thereof, and at least one heterologous nucleotide sequence encoding an immunogenic polypeptide .Iadd.inserted into the PAV-3 insertion site located in a region selected from the group consisting of the E1 region, and the E4 region of the PAV-3genome wherein the insertion renders the PAV-3 replication defective.Iaddend., and a pharmaceutically acceptable excipient.
44. The vaccine of claim 43 wherein said immunogenic polypeptide is a pathogen antigen. |
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