# US 5,753,235 A · Recombinant canine herpesviruses > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 5,753,235 A - Document ID: US5753235A - Status: Granted - Kind: A - Application number: 08/602,010 - Filed: 1996-02-15 - Granted: 1998-05-19 - Earliest claimed: 1996-02-15 - Assignees: Heska Corporation - Inventors: Elizabeth J. Haanes; Rexann S. Frank - Web page: https://patentgrep.com/patent/US5753235A ## CPC - C12N 15/86: Viral vectors - A61P 31/00: Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics - A61P 31/04: Antibacterial agents - A61P 31/10: Antimycotics - A61P 31/22: for herpes viruses - A61P 33/00: Antiparasitic agents - A61P 37/00: Drugs for immunological or allergic disorders - C07K 14/005: from viruses - C12N 7/00: Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof (preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00) - C12N 9/1211: {Thymidine kinase (2.7.1.21)} - C12N 2710/16721: Viruses as such, e.g. new isolates, mutants or their genomic sequences - C12N 2710/16722: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes - C12N 2710/16743: viral genome or elements thereof as genetic vector - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) ## Abstract The present invention includes novel recombinant canine herpes virus (CHV) and novel recombinant CHV genomes, and particularly to those CHV and CHV genomes that contain heterologous nucleic acid molecules. The present invention also relates to the use of such genomes and viruses in a variety of applications, including as therapeutic compositions to protect animals from disease. The present invention also relates to novel isolated CHV nucleic acid molecules, to CHV proteins encoded by such nucleic acid molecules, and to antibodies raised against such CHV proteins as well as to the use of such CHV nucleic acid molecules, proteins and antibodies as therapeutic compositions to protect an animal from CHV. The present invention also includes constructs comprising CHV nucleic acid molecules that include heterologous nucleic acid molecules, to recombinant vectors including such constructs, and to the use of such constructs and vectors in the production of recombinant CHV and recombinant CHV genomes. ## Claims ### Claim 1 An isolated canine herpesvirus (CHV) nucleic acid molecule that hybridizes under stringent hybridization conditions with a CHV nucleic acid region selected from the group consisting of a CdUPTase gene, a CgE gene, a CgG gene, a CgI gene, a CPK gene, a CTK gene, a CIR6 gene, a CUS2 gene, a CUS9 gene, a CUL49 gene, a CUL51 gene, a CUL48 gene, a CUL52 gene and a region of the CHV genome spanning from the 3' end of the coding region of the CUL41 gene through the 3' end of the coding region of the CUL38 gene. ### Claim 2 (depends on 1) The CHV nucleic acid molecule of claim 1, wherein said CdUTPase gene comprises nCdUTP.sub.459, wherein said CgE gene comprises nCgE.sub.750, wherein said CgG gene comprises nCgG.sub.1248, wherein said CgI gene comprises nCgI.sub.161, wherein said CPK gene comprises nCPK.sub.1203, wherein said CTK gene comprises nCTK.sub.280, wherein said CIR6 gene comprises nCIR6.sub.552, wherein said CUS2 gene comprises nCUS2.sub.1176, wherein said CUS9 gene comprises nCUS9.sub.579, wherein said CUL49 gene is included in nCHin.sub.3000, wherein said CUL51 gene comprises nCUL51.sub.261, wherein said CUL48 gene comprises nCUL48.sub.294, and wherein said CUL52 gene comprises nCUL52.sub.146. ### Claim 3 (depends on 1) The CHV nucleic acid molecule of claim 1, wherein said CHV nucleic acid molecule comprises a nucleic acid sequence that is at least 80% identical to a nucleic acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:35, SEQ ID NO:37, and SEQ ID NO:38. ### Claim 4 (depends on 1) The CHV nucleic acid molecule of claim 1, wherein said nucleic acid molecule hybridizes under stringent hybridization conditions with a nucleic acid molecule selected from the group consisting of nCIR6.sub.552, nCUS2.sub.1176, nCPK.sub.1203, nCgG.sub.1248, nCgI.sub.161, nCgE750, nCUS9.sub.579, nCdUTP/nCUL51.sub.743, nCTK.sub.280, nCUL48.sub.294, and nCUL52.sub.146. ### Claim 5 (depends on 1) The CHV nucleic acid molecule of claim 1, wherein said CHV nucleic acid molecule comprises a CHV nucleic acid molecule selected from the group consisting of nCAsc.sub.9300, nCAsc.sub.10000, nCHin.sub.3000, nCHin.sub.1900, nCUS.sub.5495, nCIR6.sub.552, nCUS2.sub.1176, nCPK.sub.1203, nCgG.sub.1248, nCdUTP/CUL51.sub.743, nCdUTP.sub.459, nCUS9.sub.579, nCUS9.sub.450, nCUL48.sub.294, nCUL48.sub.291, nCUL52.sub.146, nCUL52.sub.144, nCgI.sub.161, nCgI.sub.159, nCgE.sub.750, nCTK.sub.280, nCTK.sub.279, nCUL51.sub.261, and allelic variants of said CHV nucleic acid molecules. ### Claim 6 (depends on 1) The CHV nucleic acid molecule of claim 1, wherein said CHV nucleic acid molecule is selected from the group consisting of: a CHV nucleic acid molecule comprising a nucleic acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:35, SEQ ID NO:37, and SEQ ID NO:38; and a nucleic acid molecule comprising an allelic variant of any of said CHV nucleic acid molecules. ### Claim 7 (depends on 1) A recombinant molecule comprising a CHV nucleic acid molecule as set forth in claim 1 operatively linked to a transcription control sequence. ### Claim 8 A recombinant virus comprising an isolated CHV nucleic acid region selected from the group consisting of a CdUTPase gene, a CgE gene, a CgG gene, a CgI gene, a CPK gene, a CTK gene, a CIR6 gene, a CUS2 gene, a CUS9 gene, a CUL49 gene, a CUL51 gene, a CUL48 gene, a CUL52 gene and a region of the CHV genome spanning from the 3' end of the coding region of the CUL41 gene through the 3' end of the coding region of the CUL38 gene and portions thereof, wherein a portion encodes an epitope that elicits an immune response against a CHV protein. ### Claim 9 (depends on 1) A recombinant cell comprising a CHV nucleic acid molecule as set forth in claim 1. ### Claim 10 A recombinant vector comprising an isolated CHV nucleic acid region selected from the group consisting of a CdUTPase gene, a CgE gene, a CgG gene, a CgI gene, a CPK gene, a CTK gene, a CIR6 gene, a CUS2 gene, a CUS9 gene, a CUL49 gene, a CUL51 gene, a CUL48 gene, a CUL52 gene and a region of the CHV genome spanning from the 3' end of the coding region of the CUL41 gene through the 3' end of the coding region of the CUL38 gene and portions thereof, wherein the portion forms a stable hybrid with the complementary sequence of one of said regions. ### Claim 11 (depends on 10) A cell comprising said recombinant vector of claim 10. ### Claim 12 A recombinant CHV comprising a recombinant CHV genome, said CHV genome comprising a heterologous nucleic acid molecule operatively linked to a transcription control region, said heterologous nucleic acid molecule being in a region of said CHV genome selected from the group consisting of a nonessential gene of said CHV genome and an intergenic region of said CHV genome, said CHV being reproduction competent. ### Claim 13 (depends on 12) The CHV of claim 12, wherein said CHV comprises a recombinant CHV genome having an inactive gene. ### Claim 14 (depends on 13) The CHV of claim 13, wherein said inactive gene comprises an alteration selected from the group consisting of a deletion of one or more nucleotides, an insertion of one or more nucleotides, an inversion of one or more nucleotides, and a substitution of one or more nucleotides. ### Claim 15 (depends on 13) The CHV of claim 13, wherein said inactive gene comprises said heterologous nucleic acid molecule. ### Claim 16 (depends on 12) The CHV of claim 12, wherein said CHV is selected from the group consisting of a CdUTPase negative CHV, a CgC negative CHV, a CgE negative CKV, a CgG negative CEV, a CgI negative CHV, a CPk negative CHV, a CTK negative CHV, a CIR6 negative CHV, a CUS2 negative CHV, a CUS9 negative CkIv, a CUL49 negative CHV and a CUL45 negative CHV. ### Claim 17 (depends on 16) The CHV of claim 16, wherein said CHV is attenuated. ### Claim 18 (depends on 12) The CHV of claim 12, wherein said CHV comprises a recombinant CHV genome having a heterologous nucleic acid molecule in a CHV gene of said CHV genome, wherein said CHV gene is selected from the group consisting of a CdUTPase gene, a CgC gene, a CgE gene, a CgG gene, a CgI gene, a CPK gene, a CTK gene, a CIR6 gene, a CUS2 gene, a CUS9 gene, a CUL49 gene, and a CUL45 gene. ### Claim 19 (depends on 12) The CHV of claim 12, wherein said CHV comprises a recombinant CHV genome having a heterologous nucleic acid molecule in a region of said CHV genome spanning from the 3' end of the coding region of the CUL41 gene through the 3' end of the coding region of the CUL38 gene. ### Claim 20 (depends on 12) The CHV of claim 12, wherein said CHV comprises a recombinant CHV genome having a heterologous nucleic acid molecule in a naturally-occurring AscI restriction endonuclease site wit in said CHV genome. ### Claim 21 (depends on 12) The CHV of claim 12, wherein said CHV comprises a recombinant CHV genome having a heterologous nucleic acid molecule in the US region of said CHV genome at a position selected from the group consisting of a non-essential gene in said US region an a intergenic site in said US region. ### Claim 22 (depends on 12) The CHV of claim 12, wherein said CHV comprises a recombinant CHV genome having a heterologous nucleic acid molecule in a region of said CHV genome selected from the group consisting of nCAsc.sub.9300, nCAsc.sub.10000, nCHin.sub.3000, nCHin.sub.1900, and allelic variants of aid regions, wherein said heterologous nucleic acid molecule does not abrogate growth of said CHV in tissue culture. ### Claim 23 (depends on 12) The CHV of claim 12, wherein said CHV comprises a recombinant CHV genome having a heterologous nucleic acid molecule in a region of said CHV genome selected from the group consisting of US region comprising nCUS.sub.5495, a CHV UL region spanning CgC and CUL45 comprising nCgC/CUL45.sub.2100, a CgE gene comprising nCgE.sub.750, a CgI gene comprising nCgI.sub.161, a CUS9 gene comprising nCUS9.sub.579, a CHV UL region spanning CdUTPase and CUL51 comprising ncdUTP/CUL51.sub.743, a CTK gene comprising nCTK.sub.280, a CUL49 gene included in nCHin.sub.3000, and allelic variants of said regions, wherein said heterologous nucleic acid molecule does not abrogate growth of said CHV in tissue culture. ### Claim 24 (depends on 12) The CHV of claim 12, wherein said CHV comprises a recombinant CHV genome having a heterologous nucleic acid molecule in a region of said CHV genome selected from the group consisting of nCUS.sub.5495, nCgC/CUL45.sub.2100, nCgE.sub.750, nCgI.sub.161, nCUS9.sub.579,-nCdUTP/CUL51.sub.743, nCTK.sub.280, and allelic variants of said regions, wherein said heterologous nucleic acid molecule does not abrogate growth of said CHV in tissue culture. ### Claim 25 (depends on 12) The CHV of claim 12, wherein said CHV comprises a recombinant CHV genome having a heterologous nucleic acid molecule in a region of said CHV genome represented by a nucleic acid sequence comprising a nucleic acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:35, SEQ ID NO:37, SEQ ID NO:38, and allelic variants of said regions, wherein said heterologous nucleic acid molecule does not abrogate growth of said CHV in tissue culture. ### Claim 26 (depends on 12) The CHV of claim 12, wherein said heterologous nucleic acid molecule encodes a compound selected from the group consisting of orateins and RNA species. ### Claim 27 (depends on 26) The CHV of claim 26, wherein said compound is derived from a infectious agent selected from the group consisting of protozoan parasites, helminth parasites, ectoparasites, fungi bacteria, and viruses. ### Claim 28 (depends on 12) The CHV of claim 12, wherein said heterologous nucleic acid molecule comprises a gene encoding an immunomodulator. ### Claim 29 (depends on 12) The CHV of claim 12, wherein said CHV infects dogs. ### Claim 30 A recombinant CHV genome comprising a heterologous nucleic acid molecule operatively linked to a transcription control region said heterologous nucleic acid molecule being in a region of said CHV genome selected from the group consisting of a nonessential gene of said CHV genome and an intergenic region of said CHV genome, said CHV genome is reproduction competent. ### Claim 31 (depends on 30) The CHV genome of claim 30, wherein said CHV genome comprises an inactive gene. ### Claim 32 (depends on 31) The CHV genome of claim 31, wherein said inactive gene comprises an alteration selected from the group consisting of a deletion of one or more nucleotides, an insertion of one or more nucleotides, an inversion of one or more nucleotides, and a substitution of one or more nucleotides. ### Claim 33 (depends on 31) The CHV genome of claim 31, wherein said inactive gene comprises said heterologous nucleic acid molecule. ### Claim 34 (depends on 30) The CHV genome of claim 30, wherein said CHV genome is selected from the group consisting of a CdUTPase negative CHV genome, a CgG negative CHV genome, a CgE negative CHV genome, a CgG negative CHV genome, a CgI negative CHV genome, a CPK negative CHV genome, a CTK negative CHV genome, a CIR6 negative CHV genome a CUS2 negative CHV genome, a CUS9 negative CHV genome, a CUL49 negative CHV genome, and a CUL45 negative CHV genome. ### Claim 35 (depends on 30) The CHV genome of claim 30, wherein said heterologous nucleic acid molecule is in a CHV gene selected from the group consisting of a CdUTPase gene, a CgC gene, a CgE gene, a CgG gene, a CgI gene, a CPK gene, a CTK gene, a CIR6 gene, a CUS2 gene, a CUS9 gene, a CUL49 gene, and a CUL45 gene. ### Claim 36 (depends on 30) The CHV genome of claim 30, wherein said heterologous nucleic acid molecule is in a region of said CHV genome spanning from the 3' end of the coding region of the CUL41 gene through the 3' end of the coding region of the CUL38 gene. ### Claim 37 (depends on 30) The CHV genome of claim 30, wherein said CHV genome comprises a heterologus nucleic acid molecule in a naturally-occurring AscI restriction endonuclease site in raid CHV genome. ### Claim 38 (depends on 30) The CHV genome of claim 30, wherein said CHV genome comprises a heterologous nucleic acid molecule in the US region of said CHV genome at a position selected from the group consisting of a non-essential gene in said US region and an intergenic site in said US region. ### Claim 39 (depends on 30) The CHV genome of claim 30, wherein said CEV genome comprises a heterologous nucleic acid molecule in a region of said CHV genome selected from the group consisting of nCAsc.sub.9300, nCAsc.sub.10000, nCHin3000, nCHin.sub.1900, and allelic variants of said regions, wherein said heterologous nucleic acid molecule does not abrogate growth of said CHV in tissue culture. ### Claim 40 (depends on 30) The CHV genome of claim 30, wherein said CHV genome comprises a heterologous nucleic acid molecule in a region of said CHV genome selected from the group consisting of a CHV US region comprising nCUS.sub.5495, a CHV UL region spanning CgC and CUL45 comprising /CUL45.sub.2100, a CgE gene comprising nCgE.sub.750, a CgI gene comprising nCgI.sub.161, a CUS9 gene comprising nCUS9.sub.579, a CHV UL region spanning CdUTPase and CUL51 comprising nCdUTP/CUL51.sub.743, a CTK gene comprising nCTK.sub.280, a CUL49 gene included in nCHin.sub.3000 and allelic variants of said regions, wherein said heterologous nucleic acid molecule does not abrogate growth of said CHV in tissue culture. ### Claim 41 (depends on 30) The CHV genome of claim 30, wherein said CHV genome comprises a heterologous nucleic acid molecule in a region of said CHV genome Selected from the group consisting of nCUS.sub.5495, nCgC/CUL45.sub.2100, nCgE.sub.750, nCgI.sub.161, nCUS9.sub.579, nCdUTP/CUL51.sub.743, nCTK.sub.280, and allelic variants of said regions, wherein said heterologous nucleic acid molecule does not abrogate growth of said CHV in tissue culture. ### Claim 42 (depends on 30) The CHV genome of claim 30, wherein said CHV genome comprises a heterologous nucleic acid molecule in a region of said CHV genome represented by a nucleic acid sequence comprising a nucleic acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:29, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:35, SEQ ID NO:37, SEQ ID NO:38, and allelic variants said regions, wherein said heterologous nucleic acid molecule does not abrogate growth of said CHV in tissue culture. ### Claim 43 (depends on 30) The CHV genome of claim 30, wherein said heterologous nucleic acid molecule encodes a compound selected from the group consisting of proteins and RNA species. ### Claim 44 (depends on 43) The CHV genome of claim 43, wherein said compound is derived from an infectious agent selected from the group consisting of protozoan parasites, helminth parasites, ectoparasites, fungi, bacteria, and viruses. ### Claim 45 (depends on 30) The CHV genome of claim 30, wherein said heterologous nucleic acid molecule comprises a gene encoding an immunomodulator. ### Claim 46 (depends on 30) A transfected cell comprising a CHV genome as set forth in claim 30. ### Claim 47 (depends on 30) The CHV genome of claim 30, wherein said CHV genome comprises a recombinant dog herpesvirus genome. ### Claim 48 A composition for delivery to a canid, said composition comprising a recombinant CHV comprising a recombinant CHV genome, said CHV genome comprising a heterologous nucleic acid molecule operatively linked to a transcription control region, said heterologous nucleic acid molecule being in a region of said CHV genome selected from the group consisting of a nonessential gene of said CHV genome and an intergenic region of said CHV genome, said CHV being reproduction competent. ### Claim 49 (depends on 48) The composition of claim 48, wherein said composition further comprises a component selected from the group consisting of an excipient, an adjuvant, and a carrier. ### Claim 50 A composition for delivery to a canid, said composition comprising a recombinant CHV genome, said CHV genome comprising a recombinant CHV genome, said CHV genome comprising a heterologous nucleic acid molecule operatively linked to a transcription control region, said heterologous nucleic acid molecule being in a region of said CHV genome selected from the group consisting of a nonessential gene of said CHV genome and an intergenic region of said CHV genome, wherein a recombinant CHV comprising said CHV genome is reproduction competent. ### Claim 51 (depends on 50) The composition of claim 50, wherein said composition further comprises a component selected from the group consisting of an excipient, an adjuvant, and a carrier. ### Claim 52 A method to deliver a compound to a canid, said method comprising administering to said canid a composition comprising a recombinant CHV comprising a recombinant CHV genome, said CHV genome comprising a heterologous nucleic acid molecule encoding said compound, operatively linked to a transcription control region, said heterologous nucleic acid molecule being in a region of said CHV genome selected from the group consisting of a nonessential gene of said CHV genome and an intergenic region of said CHV genome, said CHV being reproduction competent. ### Claim 53 (depends on 52) The method of claim 52, wherein said composition further comprises a component selected from the group consisting of an excipient, an adjuvant, and a carrier. ### Claim 54 A method to deliver a compound to a canid, said method comprising administering to said canid a composition comprising a recombinant CHV genome, said CHV genome comprising a heterologous nucleic acid molecule encoding said compound, operatively linked to a transcription control region, said heterologous nucleic acid molecule being in a region of said CHV genome selected from the group consisting of a nonessential gene of said CHV genome and an intergenic region of said CHV genome, wherein a recombinant CHV comprising said CHV genome is reproduction competent. ### Claim 55 (depends on 54) The method of claim 54, wherein said composition further comprises a component selected from the group consisting of an excipient, an adjuvant, and a carrier. ### Claim 56 (depends on 1) A composition for delivery to a canid, said composition comprising an isolated CHV nucleic acid molecule as set forth in claim 1. ### Claim 57 (depends on 56) The composition of claim 56, wherein said composition further comprises a component selected from the group consisting of excipient, an adjuvant, and a carrier.