RECOMBINANT SWINEPOX VIRUS

Abstract: The present invention relates to a recombinant swinepox virus capable of replication comprising foreign DNA inserted into a site in the swinepox viral DNA which is not essential for replication of the swinepox virus. The invention further relates to homology vectors which produce recombinant swinepox viruses by inserting foreign DNA into swinepox viral DNA.

Description: RECOMBINANT SWINEPOX VIRUS ~ι 5 Within this application several publications are referenced by arabic numerals within parentheses. Full citations for these publications may be found at the end of the specification immediately preceding the claims. 10 The disclosures of these publications are hereby incorporated by reference into this application in order to more fully describe the state of the art to which this invention pertains. 15 Background of the Invention Swinepox virus (SPV) belongs to the family Poxviridae. Viruses belonging to this group are large, double- stranded DNA viruses that characteristically develop in 20 the cytoplasm of the host cell. SPV is the only member of the genus Suipoxviruε . Several features distinguish SPV from other poxviruses. SPV exhibits species specificity (18) compared to other poxviruses such as vaccinia which exhibit a broad host range. SPV infection 25 of tissue culture cell lines also differs dramatically from other poxviruses (24) . It has also been demonstrated that SPV does not exhibit antigenic cross- reactivity with vaccinia virus and shows no gross detectable homology at the DNA level with the ortho, 30 lepori, avi or entomopox virus groups (24) . Accordingly, what is known and described in the prior art regarding other poxviruses does not pertain a priori to swinepox virus. 35 SPV is only mildly pathogenic, being characterized by a self-limiting infection with lesions detected only in the HEET εkin and regional lymph nodes. Although the SPV infection is quite limited, pigs which have recovered from SPV are refractory to challenge with SPV, indicating development of active immunity (18) . The present invention concerns the use of SPV as a vector for the delivery of vaccine antigens and therapeutic agents to swine. The following properties of SPV support this rationale: SPV is only mildly pathogenic in swine, SPV is species specific, and SPV elicits a protective immune response. Accordingly, SPV is an excellent candidate for a viral vector delivery system, having little intrinsic risk which must be balanced against the benefit contributed by the vector's vaccine and therapeutic properties. The prior art for this invention stems first from the ability to clone and analyze DNA while in bacterial plasmids. The techniques that are available are detailed for the most part in Maniatis et al., 1983 and Sambrook et al . , 1989. These publications teach state of the art general recombinant DNA techniques. Among the poxviruses, five (vaccinia, fowlpox, canarypox, pigeon, and raccoon pox) have been engineered, previous to this disclosure, to contain foreign DNA sequences. Vaccinia virus has been used extensively to vector foreign genes (25) and is the subject of U.S. Patents 4,603,112 and 4,722,848. Similarly, fowlpox has been used to vector foreign genes and is the subject of several patent applications EPA 0 284 416, PCT WO 89/03429, and PCT WO 89/12684. Raccoon pox (10) and Canarypox (31) have been utilized to express antigens from the rabies virus. These examples of insertions of foreign genes into poxviruses do not include an example from the genus Suipoxv±rus . Thus, they do not teach methods to genetically engineer swinepox viruses, that is, where to make insertions and how to get expression in swinepox virus. The idea of using live viruses as delivery systems for antigens has a very long history going back to the first live virus vaccines. The antigens delivered were not foreign but were naturally expressed by the live virus in the vaccines. The use of viruses to deliver foreign antigens in the modern sense became obvious with the recombinant vaccinia virus studies. The vaccinia virus was the vector and various antigens from other disease causing viruses were the foreign antigens, and the vaccine was created by genetic engineering. While the concept became obvious with these disclosures, what was not obvious was the answer to a more practical question of what makes the best candidate virus vector. In answering this question, details of the pathogenicity of the virus, its site of replication, the kind of immune response it elicits, the potential it has to express foreign antigens, its suitability for genetic engineering, its probability of being licensed by regulatory agencies, etc, are all factors in the selection. The prior art does not teach these questions of utility. The prior art relating to the use of poxviruses to deliver therapeutic agents relates to the use of a vaccinia virus to deliver interleukin-2 (12) . In this case, although the interleukin-2 had an attenuating effect on the vaccinia vector, the host did not demonstrate any therapeutic benefit. The therapeutic agent that is delivered by a viral vector of the present invention must be a biological molecule that is a by-product of swinepox virus replication. This limits the therapeutic agent in the first analysis to either DNA, RNA or protein. There are examples of therapeutic agents from each of these classes of compounds in the form of anti-sense DNA, anti-sense RNA (16), ribozymes (34), suppressor tRNAs (2), interferon- inducing double stranded RNA and numerous examples of protein therapeutics, from hormones, e.g., insulin, to lymphokines, e.g., interferons and interleukins, to natural opiates. The discovery of these therapeutic agents and the elucidation of their structure and function does not make obvious the ability to use them in a viral vector delivery system. Summary of the Invention The invention provides a recombinant swinepox virus capable of replication which comprises swinepox viral DNA and foreign DNA encoding RNA which does not naturally occur in an animal into which the recombinant swinepox virus is introduced. The foreign DNA is inserted into the swinepox viral DNA at a site which is not essential for replication of the swinepox virus and is under the control of a promoter. This invention provides a homology vector for producing a recombinant swinepox virus by inserting foreign DNA into the genomic DNA of a swinepox virus which comprises a double-stranded DNA molecule. This molecule consists essentially of double-stranded foreign DNA encoding RNA which does not naturally occur in an animal into which the recombinant swinepox virus is introduced. At one end of this foreign DNA is double-stranded swinepox viral DNA homologous to genomic DNA located at one side of a site on the genomic DNA which is not essential for replication of the swinepox virus. At the other end of the foreign DNA is double-stranded swinepox viral DNA homologous to genomic DNA located at the other side of the same site on the genomic DNA. Brief Description of the Figures Figure 1 Details of the SPV Kasza Strain. Diagram of SPV genomic DNA showing the unique long and Terminal repeat (TR) regions. A restriction map for the enzyme HindΣII is indicated (23) . Fragments are lettered in order of decreasing size. Note that the terminal repeats are greater than 2.1 kb but less than 9.7 kb in size. Figure 2 DNA sequence from homology vector 515-85.1. The sequence of two regions of the homology vector 515-85.1 are shown. The first region (Figure 2A) (SEQ ID N0:1) covers a 599 base pair sequence which flanks the unique Accl site as indicated in Figure 3. The beginning (Met) and end (Val) of a 115 amino acid ORF is indicated by the translation of amino acids below the DNA sequence. The second region (Figure 2B) (SEQ ID NO:3) covers the 899 base pairs upstream of the unique indlll site as indicated in Figure 3. The beginning (Asp) and end (lie) of a 220 amino acid ORF is indicated by the translation of amino acids below the DNA sequence. Figure 3 Homology between the 515.85.1 ORF and the Vaccinia virus OIL ORF. The first line shows a restriction map of the SPV HindlZl M fragment. The second map shows a restriction map of the DNA insertion in plasmid 515-85.1. The location of the 515-85.1 [W OIL-like] ORF is indicated on the map. The locations of the DNA sequences shown in Figure 2 are indicated below the map by heavy bars. The third line shows the homolo between the W OIL ORF SE ID NO:5) and the 515-85.1 ORF (SEQ ID NO:6) at their respective N-termini. The fourth line shows the homology between the W OIL ORF (SEQ ID NO:7) and the 515-85.1 ORF (SEQ ID NO:8) at their respective C-termini. Figure 4 Detailed description of the DNA insertion in Homology Vector 520-17.5. Diagram showing the orientation of DNA fragments assembled in plasmid 520-17.5. The origin of each fragment is indicated in the table. The sequences located at each of the junctions between fragments is also shown (SEQ ID NO , s: 9, 10, 13, and 16). The restriction sites used to generate each fragment as well as the synthetic linker sequences which were used to join the fragments are described for each junction. The synthetic linker sequences are underlined by a heavy bar. The location of several gene coding regions and regulatory elements are also given. The following two conventions are used: numbers in parenthesis () refer to amino acids, and restriction sites in brackets [] indicate the remnants of sites which were destroyed during construction. The following abbreviations are used, swinepox virus (SPV) , early promoter l (EP1) , late promoter 2 (LP2) , lactose operon Z gene (lacZ) , and Escherichia coli (E. coli) . Figure 5 Detailed description of the DNA insertion in Homology Vector 538-46.16. Diagram showing the orientation of DNA fragments assembled in plasmid 538-46.16. The origin of each fragment is indicated in the table. The sequences located at each of the junctions between fragments are also shown (SEQ ID NO's: 17, 18, 21, 26, and 28). The restriction sites used to generate each fragment as well as the synthetic linker sequences which, were used to join the fragments are described for each junction. The synthetic linker sequences are underlined by a heavy bar. The location of several gene coding regions and regulatory elements is also given. The following two conventions are used: numbers in parenthesis () refer to amino acids, and restriction sites in brackets [] indicate the remnants of sites which were destroyed during construction. The following abbreviations are used, swinepox virus (SPV) , pseudorabies virus (PRV) , glycoprotein 50 (gp50) , glycoprotein 63 (gp63), early promoter 1 (EP1) , late promoter 1 (LP1) , late promoter 2 (LP2), lactose operon Z gene (lacZ) , and Escherichia coli (E. coli) . Figure 6 Western blot of lysates from recombinant SPV infected cells with anti-serum to PRV. Lanes (A) uninfected Vero cell lysate, (B) S-PRV-000 (pseudorabies virus Cooper strain) infected cell lysate, (C) pre-stained molecular weight markers, (D) uninfected EMSK cell lysate, (E) S-SPV-000 infected cell lysate, (F) S-SPV-003 infected cell lysate, (G) S-SPV-008 infected cell lysate. Cell lysates were prepared as described in the PREPARATION OF INFECTED CELL LYSATES. Approximately 1/5 of the total lysate sample was loaded in each lane. Figure 7 DNA sequence of NDV Hemagglutinin-Neura inidase gene (HN) (SEQ ID NO: 29) . The sequence of 1907 base pairs of the NDV HN cDNA clone are shown. The translational start and stop of the HN gene is indicated by the amino acid translation below the DNA sequence. Figure 8 Detailed description of the DNA insertion in Homology Vector 538-46.26. Diagram showing the orientation of DNA fragments assembled in plasmid 538-46.26. The origin of each fragment is indicated in the table. The sequences located at each of the junctions between fragments is also shown (SEQ ID NO's: 31, 32, 34, 37, and 40). The restriction sites used to generate each fragment as well as the synthetic linker sequences which were used to join the fragments are described for each junction. The synthetic linker sequences are underlined by a heavy bar. The location of several gene coding regions and regulatory elements is also given. The following two conventions are used: numbers in parenthesis () refer to amino acids, and restriction sites in brackets [] indicate the remnants of sites which were destroyed during construction. The following abbreviations are used, swinepox virus (SPV) , Newcastle Disease virus (NDV) , hemagglutinin- neuraminidase (HN) , early promoter 1 (EP1) , late promoter 1 (LP1) , late promoter 2 (LP2) , lactose operon Z gene (lacZ) , and Escherichia coli (E. coli) . Detailed Description Of The Invention The present invention provides a recombinant swinepox virus (SPV) capable of replication in an animal into which the recombinant swinepox virus is introduced which comprises swinepox viral DNA and foreign DNA encoding RNA which does not naturally occur in the animal into which the recombinant swinepox virus is introduced, the foreign DNA being inserted into the swinepox viral DNA at an insertion site which is not essential for replication of the swinepox virus and being under the control of a promoter. For purposes of this invention, "a recombinant swinepox virus capable of replication" is a live swinepox virus which has been generated by the recombinant methods well known to those of skill in the art, e.g., the methods set forth in HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT SPV in Materials and Methods and has not had genetic material essential for the replication of the recombinant swinepox virus deleted. For purposes of this invention, "an insertion site which is not essential for replication of the swinepox virus" is a location in the genome where a sequence of DNA is not necessary for viral replication, for example, complex protein binding sequences, sequences which code for reverse transcriptase or an essential glycoprotein, DNA sequences necessary for packaging, etc. For purposes of this invention, a "promoter" is a specific DNA sequence on the DNA molecule to which the foreign RNA polymerase attaches and at which transcription of the foreign RNA is initiated. The invention further provides foreign RNA which encodes a polypeptide. Preferably, the polypeptide is antigenic in the animal. -11- Preferably, this antigenic polypeptide is a linear polymer of more than 10 amino acids linked by peptide bonds which stimulates the animal to produce antibodies. The invention further provides an insertion site present within the larger Hindlll to Bglll subfragment of the Hindlll M fragment of swinepox viral DNA. Preferably, the insertion site is within an open reading frame contained in the Hindlll to Bglll subfragment. Preferably, the insertion site is the keel restriction endonuclease site located in the Hindlll to Bglll subfragment. The invention further provides an insertion site within an open reading frame encoding swinepox thymidine kinase. For purposes of this invention, an "open reading frame" is a segment of DNA which contains codons that can be transcribed into RNA which can be translated into an amino acid sequence and which does not contain a termination codon. The invention further provides a recombinant swinepox virus comprising foreign DNA encoding RNA encoding an antigenic polypeptide which is or is from pseudorabies virus (PRV) glycoprotein 50, pseudorabies virus (PRV) glycoprotein II, Pseudorabies virus (PRV) glycoprotein III, Pseudorabies virus (PRV) glycoprotein H, Transmissible gastroenteritis (TGE) glycoprotein 195, Transmissible gastroenteritis (TGE) matrix protein, swine rotavirus glycoprotein 38, swine parvovirus capsid protein, Serpulina hydodysenteriae protective antigen, Bovine Viral Diarrhea (BVD) glycoprotein 55, Newcastle Disease Virus (NDV) hemagglutinin-neuraminidase, swine flu hemagglutinin or swine flu neuraminidase. Preferably, the antigenic polypeptide is Pseudorabies protein is Newcastle Disease Virus (NDV) hemagglutinin- neuraminidase. The invention further provides a recombinant swinepox virus comprising foreign DNA encoding RNA encoding an antigenic polypeptide which is or is from Serpulina hyodysenteriae , Foot and Mouth Disease Virus, Hog Cholera Virus, Swine Influenza Virus, African Swine Fever Virus or Mycoplasma hyopneumoniae . The invention further provides a recombinant swinepox virus where the foreign DNA encodes RNA which encodes a polypeptide which is a detectable marker. Preferably the detectable marker is the polypeptide E. coli β- galactosidase. For purposes of this invention, a "polypeptide which is a detectable marker" includes the bimer, tri er and tetramer form of the polypeptide. E. coli β-galactosidase is a tetramer composed of four polypeptides or monomer sub-units. Preferably, this recombinant swinepox virus is designated S-SPV-003 (ATCC Accession No. VR 2335) . The S-SPV-003 swinepox virus has been deposited pursuant to the Budapest Treaty on the International Deposit of Microorganisms for the Purposes of Patent Procedure with the Patent Culture Depository of the American Type Culture Collection, 12301 Parklawn Drive, Rockville, Maryland 20852 U.S.A. under ATCC Accession No. VR 2335. The invention further provides for a recombinant swinepox virus capable of replication which contains foreign DNA encoding RNA encoding the antigenic polypeptide pseudorabies virus (PRV) glycoprotein 50 further comprising foreign DNA encoding a polypeptide which is a detectable marker. Preferably, this recombinant swinepox virus is designated S-SPV-008 (ATCC Accession No. VR 2339) . The S-SPV-008 swinepox virus has been deposited pursuant to the Budapest Treaty on the International De osit of Microor s Procedure with the Patent Culture Depository of the American Type Culture Collection, 12301 Parklawn Drive, Rockville, Maryland 20852 U.S.A. under ATCC Accession No. VR 2339. The invention further provides for a recombinant swinepox virus capable of replication which contains foreign DNA encoding RNA encoding the antigenic polypeptide Newcastle Disease Virus (NDV) hemagglutinin-neuraminidase further comprising foreign DNA encoding a polypeptide which is a detectable marker. Preferably, this recombinant swinepox virus is designated S-SPV-009 (ATCC Accession No. VR 2344) . The S-SPV-009 swinepox virus has been deposited pursuant to the Budapest Treaty on the International Deposit of Microorganisms for the Purposes of Patent Procedure with the Patent Culture Depository of the American Type Culture Collection, 12301 Parklawn Drive, Rockville, Maryland 20852 U.S.A. under ATCC Accession No. VR 2344. The invention further provides that the inserted foreign DNA is under the control of a promoter. Preferably, the promoter is a swinepox viral promoter. Preferably, the promoter is a synthetic pox viral promoter. For purposes of this invention, the promoters were generated by methods well known to those of skill in the art, for example, as set forth in the STRATEGY FOR THE CONSTRUCTION OF SYNTHETIC POX VIRAL PROMOTERS in Materials and Methods. For purposes of this invention, a synthetic pox promoter includes a synthetic late pox - promoter, a synthetic early pox promoter or a synthetic early/late pox promoter. The invention provides for a homology vector for producing a recombinant swinepox virus by inserting foreign DNA into the genomic DNA of a swinepox virus. The homology vector comprises a double-stranded DNA molecule consisting essentiall of a double-stranded foreign DNA encoding RNA which does not naturally occur in an animal into which the recombinant swinepox virus is introduced, with at one end of the foreign DNA, double- stranded swinepox viral DNA homologous to genomic DNA located at one side of a site on the genomic DNA which is not essential for replication of the swinepox virus, and at the other end of the foreign DNA, double-stranded swinepox viral DNA homologous to genomic DNA located at the other side of the same site on the genomic DNA. Preferably, the RNA encodes a polypeptide. In one embodiment, the polypeptide is antigenic in the animal. Preferably, the antigenic polypeptide is or is from pseudorabies virus (PRV) glycoprotein 50, pseudorabies virus (PRV) glycoprotein II, Pseudorabies virus (PRV) glycoprotein III, Pseudorabies virus (PRV) glycoprotein H, Transmissible gastroenteritis (TGE) glycoprotein 195, Transmissible gastroenteritis (TGE) matrix protein, swine rotavirus glycoprotein 38, swine parvovirus capsid protein, Serpulina hydodysenteriae protective antigen, Bovine Viral Diarrhea (BVD) glycoprotein 55, Newcastle Disease Virus (NDV) hemagglutinin-neuraminidase, swine flu hemagglutinin or swine flu neuraminidase. Preferably, the antigenic polypeptide is or is from Serpulina hyodysenteriae, Foot and Mouth Disease Virus, Hog Cholera Virus, Swine Influenza Virus, African Swine Fever Virus or Mycoplasma hyopneumoniae . In one embodiment, the polypeptide is a detectable marker. Preferably, the polypeptide which is a detectable marker is E. coli β-galactosidase. In one embodiment of the invention, the double-stranded swinepox viral DNA is homologous to genomic DNA present within the larger Hindlll to Bglll subfragment of the Hindlll M fragment of swinepox virus. Preferably, the - genomic DNA present within the open reading frame contained in this Hindlll to Bglll subfragment. Preferably, the double-stranded swinepox viral DNA is homologous to genomic DNA present within the Accl restriction endonuclease site located in this Hindlll to Bglll subfragment. For purposes of this invention, a "homology vector" is a plasmid constructed to insert foreign DNA in a specific site on the genome of a swinepox virus. In one embodiment of the invention, the double-stranded swinepox viral DNA in the homology vector is homologous to genomic DNA present within the open reading frame encoding swinepox thymidine kinase. The invention further provides a homology vector where foreign DNA further comprises a synthetic pox viral promoter. The invention further provides a vaccine which comprises an effective immunizing amount of a recombinant swinepox virus of the present invention and a suitable carrier. Suitable carriers for the pseudorabies virus are well known in the art and include proteins, sugars, etc. One example of such a suitable carrier is a physiologically balanced culture medium containing one or more stabilizing agents such as stabilized, hydrolyzed proteins, lactose, etc. For purposes of this invention, an "effective immunizing amount" of the recombinant swinepox virus of the present invention is within the range of 10 3 to 10 9 PFU/dose. The present invention also provides a method of immunizing an animal, wherein the animal is a swine, bovine, equine, caprine or ovine. For purposes of this invention, this includes immunizing the animal against the virus or viruses which cause the disease or diseases pseudorabies, transmissible gastroenteritis, swine rotavirus, swine parvovirus, Serpulina hyodysenteriae, bovine viral diarrhea, Newcastle disease, swine flu, foot and mouth disease, hog cholera, African swine fever or Mycoplasma hyopneumoniae . The method comprises administering to the animal an effective immunizing dose of the vaccine of the present invention. The vaccine may be administered by any of the methods well known to those skilled in the art, for example, by intramuscular, subcutaneous, intraperitoneal or intravenous injection. Alternatively, the vaccine may be administered intranasally or orally. The present invention also provides a method for testing a swine to determine whether the swine has been vaccinated with the vaccine of the present invention, particularly the embodiment which contains the recombinant swinepox virus S-SPV-008 (ATCC Accession No. VR 2339) , or is infected with a naturally-occurring, wild-type pseudorabies virus. This method comprises obtaining from the swine to be tested a sample of a suitable body fluid, detecting in the sample the presence of antibodies to pseudorabies virus, the absence of such antibodies indicating that the swine has been neither vaccinated nor infected, and for the swine in which antibodies to pseudorabies virus are present, detecting in the sample the absence of antibodies to pseudorabies virus antigens which are normally present in the body fluid of a swine infected by the naturally-occurring pseudorabies virus but which are not present in a vaccinated swine indicating that the swine was vaccinated and is not infected. The present invention also provides a host cell infected with a recombinant swinepox virus capable of replication. In one embodiment, the host cell is a mammalian cell. Preferably, the mammalian cell is a Vero cell. Preferably, the mammalian cell is an EMSK cell. For purposes of this invention a "host cell" is a cell used to propagate a vector and its insert. Infecting the cells was accomplished by methods well known to those of skill in the art, for example, as set forth in INFECTION - TRANSFECTION PROCEDURE in Material and Methods. Methods for constructing, selecting and purifying recombinant swinepox virus, including S-SPV-003, S-SPV- 008 and S-SPV-009, are detailed below in Materials and Methods. Materials and Methods PREPARATION OF SWINEPOX VIRUS STOCK SAMPLES. Swinepox virus (SPV) samples were prepared by infecting embryonic swine kidney (EMSK) cells at a multiplicity of infection of 0.01 PFU/cell in a 1:1 mixture of Iscove's Modified Dulbecco's Medium (IMDM) and RPMI 1640 medium containing 2 mM glutamine, 100 units/ml penicillin, 100 units/ml streptomycin (these components were obtained from Sigma or equivalent supplier, and hereafter are referred to as EMSK negative medium) . Prior to infection, the cell monolayers were washed once with EMSK negative medium to remove traces of fetal bovine serum. The SPV contained in the initial inoculum (0.5 ml for 10 cm plate; 10 ml for T175 cm flask) was then allowed to absorb onto the cell monolayer for two hours, being redistributed every half hour. After this period, the original inoculum was brought up to the recommended volume with the addition of complete EMSK medium (EMSK negative medium plus 5% fetal bovine serum) ► The plates were incubated at 37°C in 5% C0 2 until cytopathic effect was complete. The medium and cells were harvested and frozen in a 50 ml conical screw cap tube at -70°C. Upon thawing at 37°C, the virus stock was aliquoted into 1.0 ml vials and refrozen at -70°C. The titers were usually about 10 6 PFU/ml. PREPARATION OF SPV DNA. For swinepox virus DNA isolation, a confluent monolayer of EMSK cells in a T175 cm 2 flask was infected at a multiplicity of 0.1 and incubated 4-6 days until the cells were showing 100% cytopathic effect. The infected cells were then harvested by scraping the cells into the medium and centrifuging at 3000 rpm for 5 minutes in a clinical centrifuge. The medium was decanted, and the cell pellet was gently resuspended in 1.0 ml Phosphate Buffer Saline (PBS: 1.5g Na 2 HP0 4 , 0.2g KH 2 P0 4 , 0.8g NaCL and 0.2g KCl per liter H 2 0) (per T175) and subjected to two successive freeze-thaws (-70° C to 37° C) . Upon the last thaw, the cells (on ice) were sonicated two times for 30 seconds each with 45 seconds cooling time in between. Cellular debris was then removed by centrifuging (Sorvall RC-5B superspeed centrifuge) at 3000 rpm for 5 minutes in a HB4 rotor at 4° C. SPV virions, present in the supernatant, were then pelleted by centrifugation at 15,000 rpm for 20 minutes at 4° C in a SS34 rotor (Sorvall) and resuspended in 10 mM Tris (pH 7.5). This fraction was then layered onto a 36% sucrose gradient (w/v in 10 mM tris pH 7.5) and centrifuged (Beckman L8-70M Ultracentrifuge) at 18,000 rpm for 60 minutes in a SW41 rotor (Beckman) at 4° C. The virion pellet was resuspended in 1.0 ml of 10 mM tris pH 7.5 and sonicated on ice for 30 seconds. This fraction was layered onto a 20% to 50% continuous sucrose gradient and centrifuged 16,000 rpm for 60 minutes in a SW41 rotor at 4° C. The SPV virion band located about three quarters down the gradient was harvested, diluted with 20% sucrose and pelleted by centrifugation at 18,000 rpm for 60 minutes in a SW41 rotor at 4° C. The resultant pellet was then washed once with 10 mM Tris pH 7.5 to remove traces of sucrose and finally resuspended in 10 mM Tris pH 7.5. SPV DNA was then extracted from the purified virions by lysis (4 hours at 60° C) induced by the addition of EDTA, SDS, and proteinase K to final concentrations of 20 mM, 0.5% and 0.5 mg/ml, respectively. After digestion, three phenol:chloroform (1:1) extractions were conducted and the sample precipitated by the addition of two volumes of absolute ethanol and incubation at -20° C for 30 minutes. The sample was then centrifuged in an Eppendorf minifuge for 5 minutes at full speed. The supernatant was decanted, and the pellet air dried and rehydrated in 0.01 M Tris pH 7.5, 1 mM EDTA at 4° C. PREPARATION OF INFECTED CELL LYSATES. For cell lysate preparation, serum free medium was used. A confluent monolayer of cells (EMSK for SPV or VERO for PRV) in a 25 cm 2 flask or a 60 mm petri dish was infected with 100 μl of virus sample. After cytopathic effect was complete, the medium and cells were harvested and the cells were pelleted at 3000 rpm for 5 minutes in a clinical centrifuge. The cell pellet was resuspended in 250 μl of disruption buffer (2% sodium dodecyl sulfate, 2% β- mercapto-ethanol) . The samples were sonicated for 30 seconds on ice and stored at -20°C. WESTERN BLOTTING PROCEDURE. Samples of lysates and protein standards were run on a polyacrylamide gel according to the procedure of Laemnli (1970) . After gel electrophoresis the proteins were transferred and processed according to Sambrook et al . (1982). The primary antibody was a swine anti-PRV serum (Shope strain; lot370, PDV8201, NVSL, Ames, IA) diluted 1:100 with 5% non-fat dry milk in Tris-sodium chloride, and sodium Azide (TSA: 6.6lg Tris-HCl, 0.97g Tris-base, 9.0g NaCl and 2.0g Sodium Azide per liter H 2 0) . The secondary antibody was a goat anti-swine alkaline phosphatase conjugate diluted 1:1000 with TSA. MOLECULAR BIOLOGICAL TECHNIQUES. Techniques for the manipulation of bacteria and DNA, including such procedures as digestion with restriction endonucleases, gel electrophoresis, extraction of DNA from gels, ligation, phosphorylation with kinase, treatment with phosphatase, growth of bacterial cultures, transformation of bacteria with DNA, and other molecular biological methods are described by Maniatis et al . (1982) and Sambrook et al . (1989) . Except as noted, these were used with minor variation. DNA SEQUENCING. Sequencing was performed using the USB Sequenase Kit and 35 S-dATP (NEN) . Reactions using both the dGTP mixes and the dITP mixes were performed to clarify areas of compression. Alternatively, compressed areas were resolved on formamide gels. Templates were double-stranded plasmid subclones or single stranded M13 subclones, and rimers were either made to the vector just outside the insert to be sequenced, or to previously obtained sequence. Sequence obtained was assembled and compared using Dnastar software. Manipulation and comparison of sequences obtained was performed with Superclone™ and Supersee™ programs from Coral Software. CLONING WITH THE POLYMERASE CHAIN REACTION. The polymerase chain reaction (PCR) was used to introduce restriction sites convenient for the manipulation of various DNAs. The procedures used are described by Innis, et al . (1990). In general, amplified fragments were less than 500 base pairs in size and critical regions of amplified fragments were confirmed by DNA sequencing. The primers used in each case are detailed in the descriptions of the construction of homology vectors below. HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT SPV. This method relies upon the homologous recombination between the swinepox virus DNA and the plasmid homology vector DNA which occurs in the tissue culture cells containing both swinepox virus DNA and transfected plasmid homology vector. For homologous recombination to occur, the monolayers of EMSK cells are infected with S-SPV-001 (Kasza SPV strain, 17) at a multiplicity of infection of 0.01 PFU/cell to introduce replicating SPV (i.e. DNA synthesis) into the cells. The plasmid homology vector DNA is then transfected into these cells according to the INFECTION - TRANSFECTION PROCEDURE. The construction of homology vectors used in this procedure is described below INFECTION - TRANSFECTION PROCEDURE. 6 cm plates of EMSK cells (about 80% confluent) were infected with S-SPV-001 at a multiplicity of infection of 0.01 PFU/cell in EMSK negative medium and incubated at 37°C in a humidified 5% C0 2 environment for 5 hours. The transfection procedure used is essentially that recommended for Lipofectin™ Reagent (BRL) . Briefly, for each 6 cm plate, 15 μg of plasmid DNA was diluted up to 100 μl with H 2 0. Separately, 50 micrograms of Lipofectin Reagent was diluted to 100 μl with H 2 0. The 100 μl of diluted Lipofectin Reagent was then added dropwise to the diluted plasmid DNA contained in a polystyrene 5 ml snap cap tube and mixed gently. The mixture was then incubated for 15- 20 minutes at room temperature. During this time, the virus inoculum was removed from the 6 cm plates and the cell monolayers washed once with EMSK negative medium. Three ml of EMSK negative medium was then added to the plasmid DNA/lipofectin mixture and the contents pipetted onto the cell monolayer. The cells were incubated overnight (about 16 hours) at 37°C in a humidified 5% C0 2 environment. The next day the 3 ml of EMSK negative medium was removed and replaced with 5 ml EMSK complete medium. The cells were incubated at 37°C in 5% C0 2 for 3-7 days until cytopathic effect from the virus was 80-100%. Virus was harvested as described above for the preparation of virus stocks. This stock was referred to as a transfection stock and was subsequently screened for recombinant virus by the BLUOGAL SCREEN FOR RECOMBINANT SWINEPOX VIRUS OR CPRG SCREEN FOR RECOMBINANT SWINEPOX VIRUS. SCREEN FOR RECOMBINANT SPV EXPRESSING B-GALACTOSIDASE (BLUOGAL AND CPRG ASSAYS) . When the E. coli β- galactosidase (lacZ) marker gene was incorporated into a recombinant virus the plaques containing the recombinants were visualized by one of two simple methods. In the first method, the chemical Bluogal™ (Bethesda Research Labs) was incorporated (200 μg/ml) into the agarose overlay during the plaque assay, and plaques expressing active β-galactosidase turned blue. The blue plaques were then picked onto fresh cells (EMSK) and purified by further blue plaque isolation. In the second method, CPRG (Boehringer Mannheim) was incorporated (400 μg/ml) into the a arose overla durin the la ue assa and la ues expressing active β-galactosidase turned red. The red plaques were then picked onto fresh cells (EMSK) and purified by further red plaque isolation. In both cases viruses were typically purified with three rounds of plaque purification. SCREEN FOR FOREIGN GENE EXPRESSION IN RECOMBINANT SPV USING BLACK PLAQUE ASSAYS. To analyze expression of foreign antigens expressed by recombinant swinepox viruses, monolayers of EMSK cells were infected with recombinant SPV, overlayed with nutrient agarose media and incubated for 6-7 days at 37°C for plaque development to occur. The agarose overlay was then removed from the dish, the cells fixed with 100% methanol for 10 minutes at room temperature and the cells air dried. Fixation of the cells results in cytoplasmic antigen as well as surface antigen detection whereas specific surface antigen expression can be detected using non-fixed cells. The primary antibody was then diluted to the appropriate dilution with PBS and incubated on the cell monolayer for 2 hours at room temperature. To detect PRV gp50 expression from S-SPV-008, swine anti-PRV serum (Shope strain; lot370, PDV8201, NVSL, Ames, IA) was used (diluted 1:100). To detect NDV HN expression from S-SPV- 009, a rabbit antiserum specific for the HN protein (rabbit anti-NDV#2) was used (diluted 1:1000). Unbound antibody was then removed by washing the cells three times with PBS at room temperature. The secondary antibody, either a goat anti-swine (PRVgpδO; S-SPV-008) or goat anti-rabbit (NDV HN; S-SPV-009) , horseradish peroxidase conjugate was diluted 1:250 with PBS and incubated with the cells for 2 hours at room temperature. Unbound secondary antibody was then removed by washing the cells three times with PBS at room temperature. The cells were then incubated 15-30 minutes at room temperature with freshly prepared substrate solution (100 μg/ml 4-chloro-l-naphthol, 0.003% H 2 0 2 in PBS). Plaques ex ressin the correct anti en stain black. STRATEGY FOR THE CONSTRUCTION OF SYNTHETIC POX VIRAL PROMOTERS. For recombinant swinepox vectors synthetic pox promoters offer several advantages including the ability to control the strength and timing of foreign gene expression. We chose to design three promoter cassettes LP1, EP1 and LP2 based on promoters that have been defined in the vaccinia virus (1, 7 and 8) . Each cassette was designed to contain the DNA sequences defined in vaccinia flanked by restriction sites which could be used to combine the cassettes in any order or combination. Initiator methionines were also designed into each cassette such that inframe fusions could be made at either EcoRI or BamHI sites. A set of translational stop codons in all three reading frames and an early transcriptional termination signal (9) were also engineered downstream of the inframe fusion site. DNA encoding each cassette was synthesized according to standard techniques and cloned into the appropriate homology vectors (see Figures 4, 5 and 8) . HOMOLOGY VECTOR 515-85.1. The plasmid 515-85.1 was constructed for the purpose of inserting foreign DNA into SPV. It contains a unique AccI restriction enzyme site into which foreign DNA may be inserted. When a plasmid, containing a foreign DNA insert at the AccI site, is used according to the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT SPV a virus containing the foreign DNA will result. A restriction map of the DNA insert in homology vector 515-85.1 is given in figure 4. It may be constructed utilizing standard recombinant DNA techniques (22 and 29) , by joining two restriction fragments from the following sources. The first fragment is an approximately 2972 base pair Hindlll to BamHI restriction fragment of pSP64 (Promega) . The second fragment is an approximately 3300 base pair Hindlll to Bglll restriction sub-fragment of the SPV Hindlll restriction fragment M (23). HOMOLOGY VECTOR 520-17.5. The plasmid 520-17.5 was constructed for the purpose of inserting foreign DNA into SPV. It incorporates an E. coli β-galactosidase (lacZ) marker gene flanked by SPV DNA. Upstream of the marker gene is an approximately 2015 base pair fragment of SPV DNA. Downstream of the marker gene is an approximately 1103 base pair fragment of SPV DNA. When this plasmid is used according to the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT SPV a virus containing DNA coding for the marker gene will result. Note that the β- galactosidase (lacZ) marker gene is under the control of a synthetic early/late pox promoter. A detailed description of the plasmid is given in figure 4. It may be constructed utilizing standard recombinant DNA techniques (22 and 30) , by joining restriction fragments from the following sources with the synthetic DNA sequences indicated in figure 4. The plasmid vector is derived from an approximately 2972 base pair Hindlll to BamHI restriction fragment of pSP64 (Promega) . Fragment 1 is an approximately 2015 base pair Hindlll to AccI restriction sub-fragment of the SPV Hindlll restriction fragment M (23) . Fragment 2 is an approximately 3010 base pair BamHI to PvuII restriction fragment of plasmid pJF751 (11) . Fragment 3 is an approximately 1103 base pair AccI to Bglll restriction sub-fragment of the SPV Hindlll fragment M (23) . HOMOLOGY VECTOR 538-46.16. The plasmid 538-46.16 was constructed for the purpose of inserting foreign DNA into SPV. It incorporates an E . coli β-galactosidase (lacZ) marker gene and the PRV gp50 gene flanked by SPV DNA. Upstream of the foreign genes is an approximately 2015 base pair fragment of SPV DNA. Downstream of the foreign genes is an approximately 1103 base pair fragment of SPV DNA. When this plasmid is used according to the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT SPV a virus containing DNA coding for the foreign genes will result. Note that the β-galactosidase (lacZ) marker gene is under the control of a synthetic late pox promoter and the gp50 gene is under the control of a synthetic early/late pox promoter. A detailed description of the plasmid is given in figure 5. It may be constructed utilizing standard recombinant DNA techniques (22 and 30) , by joining restriction fragments from the following sources with the synthetic DNA sequences indicated in figure 5. The plasmid vector is derived from an approximately 2972 base pair Hindlll to BamHI restriction fragment of pSP64 (Promega) . Fragment 1 is an approximately 2015 base pair Hindlll to AccI restriction sub-fragment of the SPV Hindlll restriction fragment M (23) . Fragment 2 is an approximately 3010 base pair BamHI to PvuII restriction fragment of plasmid pJF751 (11) . Fragment 3 is an approximately 1571 base pair -EcoRI to StuI restriction sub-fragment of the PRV BamHI fragment 7 (21) . Note that the .EcoRI site was introduced in to this fragment by PCR cloning. In this procedure the primers described below were used along with a template consisting of a PRV BamHI #7 fragment subcloned into pSP64. The first primer 87.03 (5'- CGCGAATTCGCTCGCAGCGCTATTGGC-3') (SEQ ID NO:41) sits down on the PRV gp50 sequence (26) at approximately amino acid 3 priming toward the 3' end of the gene. The second primer 87.06 (5'-GTAGGAGTGGCTGCTGAAG-3') (SEQ ID NO:42) sits down on the opposite strand at approximately amino acid 174 priming toward the 5' end of the gene. The PCR product may be digested with coRI and Sail to produce an approximately 509 base pair fragment. The approximately 1049 base pair Sail to StuI sub-fragment of PRV BamHI #7 may then be ligated to the approximately 509 base pair EcoRI to Sail fragment to generate the approximately 1558 base pair EcoRI to StuI fragment 3. Fragment 4 is an approximately 1103 base pair AccI to Bglll restriction sub-fragment of the SPV Hindlll fragment M (23) . HOMOLOGY VECTOR 538-46.26. The plasmid 538-46.26 was constructed for the purpose of inserting foreign DNA into SPV. It incorporates an E. coli β-galactosidase (lacZ) marker gene and the Newcastle Disease Virus (NDV) hemagglutinin-Neuraminidase (HN) gene flanked by SPV DNA. Upstream of the foreign genes is an approximately 2015 base pair fragment of SPV DNA. Downstream of the foreign genes is an approximately 1103 base pair fragment of SPV DNA. When this plasmid is used according to the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT SPV a virus containing DNA coding for the foreign genes will result. Note that the β-galactosidase (lacZ) marker gene is under the control of a synthetic late pox promoter and the HN gene is under the control of a synthetic early/late pox promoter. A detailed description of the plasmid is given in figure 8. It may be constructed utilizing standard recombinant DNA techniques (22 and 30) , by joining restriction fragments from the following sources with the synthetic DNA sequences indicated in figure 8. The plasmid vector is derived from an approximately 2972 base pair Hindlll to BamHI restriction fragment of pSP64 (Promega) . Fragment 1 is an approximately 2015 base pair Hindlll to AccI restriction sub-fragment of the SPV Hindlll restriction fragment M (23) . Fragment 2 is an approximately 1810 base pair Avail to Nael restriction fragment of a NDV HN cDNA clone. The sequence of the HN cDNA clone is given in figure 7. The cDNA clone was generated from the Bl strain of NDV using standard cDNA cloning techniques (14) . Fragment 3 is an approximately 3010 base pair BamHI to PvuII restriction fragment of plasmid pJF751 (11) . Fragment 4 is an approximately 1103 base pair AccI to Bglll restriction sub-fragment of the SPV Hindlll fragment M (23) . HOMOLOGY VECTOR 520-90.15. The plasmid 520-90.15 was constructed for the purpose of inserting foreign DNA into SPV. It contains a unique Ndel restriction enzyme site into which foreign DNA may be inserted. When a plasmid, containing a foreign DNA insert at the Ndel site, is used according to the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT SPV a virus containing the foreign DNA will result. Plasmid 520-90.15 was constructed utilizing standard recombinant DNA techniques (22 and 30) , by joining two restriction fragments from the following sources. The first fragment is an approximately 2972 base pair Hindlll to BamHI restriction fragment of pSP64 (Promega) . The second fragment is an approximately 1700 base pair Hindlll to BamHI restriction subfragment of the SPV Hindlll restriction fragment G (23). Examples Example 1 Homolocry Vector 515-85.1 The homology vector 515-85.1 is a plasmid useful for the insertion of foreign DNA into SPV. Plasmid 515-85.1 contains a unique AccI restriction site into which foreign DNA may be cloned. A plasmid containing such a foreign DNA insert may be used according to the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT SPV to generate a SPV containing the foreign DNA. For this procedure to be successful it is important that the insertion site (AccI) be in a region non- essential to the replication of the SPV and that the site be flanked with swinepox virus DNA appropriate for mediating homologous recombination between virus and plasmid DNAs. We have demonstrated that the AccI site in homology vector 515-85.1 may be used to insert foreign DNA into at least three recombinant SPV (see examples 2- 4). In order to define an appropriate insertion site, a library of SPV Hindlll restriction fragments was generated. Several of these restriction fragments (Hindlll fragments G, J, and M see figure 1) were subjected to restriction mapping analysis. Two restriction sites were identified in each fragment as potential insertion sites. These sites included Hpal and Nrul in fragment G, Ball and Xbal in fragment J, and AccI and PstI in fragment M. A β-galactosidase (lacZ) marker gene was inserted in each of the potential sites. The resulting plasmids were utilized in the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT SPV. The generation of recombinant virus was determined by the SCREEN FOR RECOMBINANT SPV EXPRESSING β-GALACTOSIDASE ASSAYS. Four of the six sites were found to generate recombinant virus, however the ability of each of these viruses to be purified away from the parental SPV varied greatly. In one case virus could not be purified above the level of 1%, in another case virus could not be purified above the level of 50%, and in a third case virus could not be purified above the level of 90%. The inability to purify these viruses indicates instability at the insertion site. This makes the corresponding sites inappropriate for insertion of foreign DNA. However the insertion at one site, the AccI site of Homology vector 515-85.1, resulted in a virus which was easily purified to 100% (see example 2) , clearly defining an appropriate site for the insertion of foreign DNA. The homology vector 515-85.1 was further characterized by DNA sequence analysis. Two regions of the homology vector were seguenced. The first region covers a 599 base pair sequence which flanks the unique AccI site (see figure 2) . The second region covers the 899 base pairs upstream of the unique Hindlll site (see figure 2) . The sequence of the first region codes for an open reading frame (ORF) which shows homology to amino acids l to 115 of the vaccinia virus (W) OIL open reading frame identified by Goebel et al , 1990 (see figure 3). The sequence of the second region codes for an open reading frame which shows homology to amino acids 568 to 666 of the same vaccinia virus OIL open reading frame (see figure 3) . These data suggest that the AccI site interrupts the presumptive W OIL-like ORF at approximately amino acid 41, suggesting that this ORF codes for a gene non-essential for SPV replication. Goebel et al. suggest that the W OIL ORF contains a leucine zipper motif characteristic of certain eukaryotic transcriptional regulatory proteins, however they indicate that it is not known whether this gene is essential for virus replication. The DNA sequence located upstream of the W OIL-like ORF (see Figure 2A) would be expected to contain a swinepox viral promoter. This swinepox viral promoter will be useful as the control element of foreign DNA introduced into the swinepox genome. Example 2 S-SPV-003 S-SPV-003 is a swinepox virus that expresses a foreign gene. The gene for E. coli β-galactosidase (lacZ gene) was inserted into the SPV 515-85.1 ORF. The foreign gene (lacZ) is under the control of a synthetic early/late promoter (EP1LP2) . S-SPV-003 was derived from S-SPV-001 (Kasza strain) . This was accomplished utilizing the homology vector 520-17.5 (see Materials and Methods) and virus S-SPV-001 in the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT SPV. The transfection stock was screened by the SCREEN FOR RECOMBINANT SPV EXPRESSING β-GALACTOSIDASE (BLUOGAL AND CPRG ASSAYS) . The final result of red plaque purification was the recombinant virus designated S-SPV-003. This virus was assayed for β-galactosidase expression, purity, and insert stability by multiple passages monitored by the blue plaque assay as described in Materials and Methods. After the initial three rounds of urification all la ues observed were blue indicating that the virus was pure, stable and expressing the foreign gene. The assays described here were carried out in VERO cells as well as EMSK cells, indicating that VERO cells would be a suitable substrate for the production of SPV recombinant vaccines. S-SPV-003 has been deposited with the ATCC under Accession No. VR 2335. Example 3 S-SPV-008 S-SPV-008 is a swinepox virus that expresses at least two foreign genes. The gene for E. coli β-galactosidase (lacZ gene) and the gene for pseudorabies virus (PRV) gp50 (26) were inserted into the SPV 515-85.1 ORF. The lacZ gene is under the control of a synthetic late promoter (LP1) and the gp50 gene is under the control of a synthetic early/late promoter (EP1LP2) . S-SPV-008 was derived from S-SPV-001 (Kasza strain) . This was accomplished utilizing the homology vector 538-46.16 (see Materials and Methods) and virus S-SPV-001 in the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT SPV. The transfection stock was screened by the SCREEN FOR RECOMBINANT SPV EXPRESSING β-GALACTOSIDASE (BLUOGAL AND CPRG ASSAYS) . The final result of red plaque purification was the recombinant virus designated S-SPV-008. This virus was assayed for β-galactosidase expression, purity, and insert stability by multiple passages monitored by the blue plaque assay as described in Materials and Methods. After the initial three rounds of purification, all plaques observed were blue indicating that the virus was pure, stable and expressing the marker gene. S-SPV-008. was assayed for expression of PRV specific antigens using the BLACK PLAQUE SCREEN FOR FOREIGN GENE EXPRESSION IN RECOMBINANT SPV. Swine anti-PRV serum was shown to react specifically with S-SPV-008 plaques and not with S-SPV-009 negative control plaques. All S-SPV- 008 observed plaques reacted with the swine antiserum indicating that the virus was stably expressing the PRV foreign gene. The black plaque assay was also performed on unfixed monolayers. The SPV plaques on the unfixed monolayers also exhibited specific reactivity with swine anti-PRV serum indicating that the PRV antigen is expressed on the infected cell surface. To confirm the expression of the PRV gp50 gene product, cells were infected with SPV and samples of infected cell lysates were subjected to SDS-polyacrylamide gel electrophoresis. The gel was blotted and analyzed using the WESTERN BLOTTING PROCEDURE. The swine anti-PRV serum was used to detect expression of PRV specific proteins. As shown in figure 6, the lysate from s-SPV-008 infected cells exhibits a specific band of approximately 48 kd, the reported size of PRV gp50 (35) . PRV gp50 is the gD homologue of HSV-1 (26) . Several investigators have shown that W expressing HSV-1 gD will protect mice against challenge with HSV-l (6 and 34) . Therefore the S-SPV-008 should be valuable as a vaccine to protect swine against PRV disease. It is anticipated that several other PRV glycoproteins will be useful in the creation of recombinant swinepox vaccines to protect against PRV disease. These PRV glycoproteins include gpll (28) , gpIII (27) , and gpH (19) . The PRV gpIII coding region has been engineered behind several synthetic pox promoters. The techniques utilized for the creation of S-SPV-008 will be used to create recombinant swinepox viruses expressing all four of these PRV glycoprotein genes. Such recombinant swinepox viruses will be useful as vaccines against PRV disease. Since the PRV vaccines described here do not express PRV gpX or gpi, they would be compatible with current PRV diagnostic tests (gX HerdChek®, gl HerdChek® and ClinEase®) which are utilized to distinguish vaccinated animals from infected animals. S-SPV-008 has been deposited with the ATCC under Accession No. VR 2339. Example 4 S-SPV-009 S-SPV-009 is a swinepox virus that expresses at least two foreign genes. The gene for E. coli β-galactosidase (lacZ gene) and the gene for Newcastle's Disease virus hemagglutinin (HN) gene were inserted into the SPV 515- 85.1 ORF. The lacZ gene is under the control of a synthetic late promoter (LP1) and the HN gene is under the control of an synthetic early/late promoter (EP1LP2) . S-SPV-009 was derived from S-SPV-001 (Kasza strain) . This was accomplished utilizing the homology vector 538-46.26 (see Materials and Methods) and virus S-SPV-001 in the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT SPV. The transfection stock was screened by the SCREEN FOR RECOMBINANT SPV EXPRESSING β-GALACTOSIDASE (BLUOGAL AND CPRG ASSAYS) . The final result of red plaque purification was the recombinant virus designated S-SPV-009. This virus was assayed for β-galactosidase expression, purity, and insert stability by multiple passages monitored by the blue plaque assay as described in Materials and Methods. After the initial three rounds of purification, all plaques observed were blue indicating that the virus was pure, stable and expressing the marker gene. S-SPV-009 was assayed for expression of PRV specific antigens using the BLACK PLAQUE SCREEN FOR FOREIGN GENE EXPRESSION IN RECOMBINANT SPV. Rabbit anti-NDV HN serum was shown to react specifically with S-SPV-009 plaques and not with S-SPV-008 negative control plaques. All S- SPV-009 observed plaques reacted with the swine antiserum indicating that the virus was stably expressing the NDV foreign gene. S-SPV-009 has been deposited with the ATCC under Acession No. VR 2344) . To confirm the expression of the NDV HN gene product, cells were infected with SPV and samples of infected cell lysates were subjected to SDS-polyacrylamide gel electrophoresis. The gel was blotted and analyzed using the WESTERN BLOTTING PROCEDURE. The rabbit anti-NDV HN serum was used to detect expression of the HN protein. The lysate from S-SPV-009 infected cells exhibited a specific band of approximately 74 kd, the reported size Of NDV HN (29) . Example 5 Homoloσv Vector 520-90.15 The homology vector 520-90.15 is a plasmid useful for the insertion of foreign DNA into SPV. Plasmid 520-90.15 contains a unique Ndel restriction site into which foreign DNA may be cloned. A plasmid containing such a foreign DNA insert has been used according to the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT SPV to generate a SPV containing the foreign DNA. For this procedure to be successful, it is important that the insertion site be in a region non- essential to the replication of the SPV and that the site be flanked with swinepox virus DNA appropriate for mediating homologous recombination between virus and plasmid DNAs. The unique Ndel restriction site in plasmid 520-90.15 is located within the coding region of the SPV thymidine kinase gene (32) . Therefore, we have shown that the thymidine kinase gene of swinepox virus is non-essential for DNA replication and is an appropriate insertion site. Example 6 The development of vaccines utilizing the swinepox virus to express antigens from various disease causing microorganisms can be engineered. TRANSMISSIBLE GASTROENTERITIS VIRUS The major neutralizing antigen of the transmissible gastroenteritis virus (TGE) , glycoprotein 195, for use in the swinepox virus vector has been cloned. The clone of the neutralizing antigen is disclosed in U.S. Serial No. 078,519, filed July 27, 1987. It is contemplated that the procedures that have been used to express PRV gp50 in SPV and are disclosed herein are applicable to TGE. PORCINE PARVOVIRUS We have cloned the major capsid protein of the porcine (swine) parvovirus (PPV) for use in the swinepox virus vector. The clone of the capsid protein is disclosed in U.S. Patent No. 5,068,192 issued November 26, 1991. It is contemplated that the procedures that have been used to express PRV gp50 in SPV and are disclosed herein are applicable to PPV. SWINE ROTAVIRUS We have cloned the major neutralizing antigen of the swine rotavirus, glycoprotein 38, for use in the swinepox virus vector. The clone of glycoprotein 38 is disclosed in U.S. Patent No. 5,068,192 issued November 26, 1991. It is contemplated that the procedures that have been used to express PRV gp50 in SPV and are disclosed herein are applicable to SRV. HOG CHOLERA VIRUS The major neutralizing antigen of the bovine viral diarrhea (BVD) virus was cloned as disclosed in U.S. Serial No. 225,032, filed July 27, 1988. Since the BVD and ho cholera viruses are cross rotective 31 the BVD virus antigen has been targeted for use in the swinepox virus vector. It is contemplated that the procedures that have been used to express PRV gp50 in SPV and are disclosed herein are applicable to BVD virus. SERPULINA HYODYSENTERIAE A protective antigen of Serpulina hyodysenteriae (3) , for use in the swinepox virus vectorhas been cloned. It is contemplated that the procedures that have been used to express PRV gp50 in SPV and are disclosed herein are also applicable to Serpulina hyodysenteriae . Antigens from the following microorganisms may also be utilized to develop animal vaccines: Swine influenza virus, foot and mouth disease virus, African swine fever virus, hog cholera virus and Mycoplas a hyodysenteriae . SUBSTITUTESHEET References 1. C. Bertholet, et al . , EMBO Journal 5, 1951-1957 (1986) . 2. R. A. Bhat, et al., Nucleic Acids Research 17, 1159- 1176 (1989). 3. D. A. Boyden, et al., Infection and Immunity 57, 3808-3815 (1989) . 4. D. B. Boyle and B. E. H. Coupar, Virus -Research 10, 343-356 (1988). 5. R. M. Buller, et al . , Nature 317, 813-815 (1985). 6. K. J. Cremer, et al., Science 228, 737-739 (1985) . 7. A. J. Davidson and B. Moss, J. Mol. Biol. 210, 749- 769 (1989) . 8. A. J. Davidson and B. Moss, J. Mol. Biol. 210, 771- 784 (1989). 9. P. L. Earl, et al . , Journal of Virology 64, 2448- 2451 (1990) . 10. J. J. Esposito, et al., Virology 165, 313 (1988) . 11. F. A. Ferrari, et al., J. of Bacteriology 161, 556- 562 (1985) . 12. C. Flexner, et al., Vaccine 8, 17-21 (1990). 13. S. J. Goebel, et al . , Virology 179, 247-266 (1990) 14. U. Gubler and B. J. Hoffman, Gene 25, 263-269 (1983) . 15. M. A. Innis, et al . , PCR Protocols A Guide to Methods and Applications, 84-91, Academic Press, Inc., San Diego (1990). 16. S. Joshi, et al . , Journal of Virology. 65, 5524-5530 (1991) . 17. L. Kasza, et al . , Am . J. Vet . Res . 21, 269-273 (1960) . 18. L. Kasza, Diseases of Swine, 254-260, Ed. A.D. Leman, et al . . The Iowa State University Press, Ames, Iowa (1981). 19. B. G. Klupp and T. C. Mettenleiter, Virology 182, 732-741 (1991) . 20. U. K. Laemnli, Nature 227, 680-685 (1970). 21. B. Lo iniczi, et al . , Journal of Virology 49, 970- 979 (1984) . 22. T. Maniatis, et al . , Molecular Cloning: A Laboratory Manual , Cold Spring Harbor Press, Cold Spring Harbor, New York (1982) . 23. R. F. Massung, and R. W. Moyer, Virology 180, 347- 354 (1991) . 24. R. F. Massung, and R. W. Moyer, Virology 180, 355- 364 (1991) . 25. B. Moss, Science 252, 1662-1667 (1991). SUBSTITUTE HEET 26. E. A. Petrovskis, et al . , Journal of Virology 59, 216-223 (1986) . 27. A. K. Robbinε et al . , Journal of Virology 58, 339- 347 (1986). 28. A. K. Robbins et al . , Journal of Virology 61, 2691- 2701 (1987) . 29. A. C. R. Samson, Journal of Virology 67, 1199-1203 (1986) . 30. J. Sambrook, et al . , Molecular Cloning A Laboratory Manual Second Edition , Cold Spring Harbor Press, Cold Spring Harbor, New York (1989) . 31. Sheffy, et al., Proceedings 65th Annual Meeting of the United States Livestock Association 65, 347-353 (1961) . 32. W. M. Schnitzlein and D. N. Tripathy, Virology 181, 727-732, (1991). 33. J. Taylor, et al . , Vaccine 9, 190-193, (1991). 34. M. Wachsman, et al . , Journal of General Virology 70, 2513-2520 (1989). 35. M. W. Wathen, et al . , Journal of Virology 51, 57-62 (1984) . 36. M. Weerasinge, Journal of Virology 65, 5531-5534 (1991) . SEQUENCE LISTING (1) GENERAL INFORMATION: (i) APPLICANT: Coc ran Ph.D., Mark D Junker M.S., David E (ii) TITLE OF INVENTION: Recombinant Swinepox Virus (iii) NUMBER OF SEQUENCES: 42 (iv) CORRESPONDENCE ADDRESS: (A) ADDRESSEE: John P. White (B) STREET: 30 Rockefeller Plaza (C) CITY: New York (D) STATE: New York (E) COUNTRY: USA (F) ZIP: 10112 (v) COMPUTER READABLE FORM: (A) MEDIUM TYPE: Floppy disk (B) COMPUTER: IBM PC compatible (C) OPERATING SYSTEM: PC-DOS/MS-DOS (D) SOFTWARE: Patentin Release #1.0, Version #1.25 (vi) CURRENT APPLICATION DATA: (A) APPLICATION NUMBER: (B) FILING DATE: (C) CLASSIFICATION: (viii) ATTORNEY/AGENT INFORMATION: (A) NAME: White, John P (ix) TELECOMMUNICATION INFORMATION: (A) TELEPHONE: (212)977-9550 (B) TELEFAX: (212)664-0525 (C) TELEX: 422523 (2) INFORMATION FOR SEQ ID N0:1: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 599 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (vi) ORIGINAL SOURCE: (A) ORGANISM: Swinepox virus (B) STRAIN: Kasza (C) INDIVIDUAL ISOLATE: S-SPV-001 (vii) IMMEDIATE SOURCE: (B) CLONE: 515-85.1 (viii) POSITION IN GENOME: (B) MAP POSITION: -23.2 (C) UNITS: %G ( ix ) FEATURE : (A) NAME/KEY: CDS (B) LOCATION: 202..597 (D) OTHER INFORMATION: /partial /codon_start= 202 /function= "Potential eukaryotic transcriptional regulatory protein" /standard_name= "515-85.1 ORF" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:l: AATGTATCCA GAGTTGTTGA ATGCCTTATC GTACCTAATA TTAATATAGA GTTATTAACT GAATAAGTAT ATATAAATGA TTGTTTTTAT AATGTTTGTT ATCGCATTTA GTTTTGCTGT ATGGTTATCA TATACATTTT TAAGGCCGTA TATGATAAAT GAAAATATAT AAGCACTTAT TTTTGTTAGT ATAATAACAC A ATG CCG TCG TAT ATG TAT- CCG AAG AAC GCA Met Pro Ser Tyr Met Tyr Pro Lys Asn Ala 1 5 10 AGA AAA GTA ATT TCA AAG ATT ATA TCA TTA CAA CTT GAT ATT AAA AAA Arg Lys Val lie Ser Lys lie lie Ser Leu Gin Leu Asp lie Lys Lys 15 20 25 CTT CCT AAA AAA TAT ATA AAT ACC ATG TTA GAA TTT GGT CTA CAT GGA Leu Pro Lys Lys Tyr lie Asn Thr Met Leu Glu Phe Gly Leu His Gly 30 35 40 AAT CTA CCA GCT TGT ATG TAT AAA GAT GCC GTA TCA TAT GAT ATA AAT Asn Leu Pro Ala Cys Met Tyr Lys Asp Ala Val Ser Tyr Asp lie Asn 45 50 55 AAT ATA AGA TTT TTA CCT TAT AAT TGT GTT ATG GTT AAA GAT TTA ATA Asn lie Arg Phe Leu Pro Tyr Asn Cys Val Met Val Lys Asp Leu lie 60 65 70 AAT GTT ATA AAA TCA TCA TCT GTA ATA GAT ACT AGA TTA CAT CAA TCT 4 Asn Val lie Lys Ser Ser Ser Val lie Asp Thr Arg Leu His Gin Ser 75 80 85 90 GTA TTA AAA CAT CGT AGA GCG TTA ATA GAT TAC GGC GAT CAA GAC ATT 5 Val Leu Lys His Arg Arg Ala Leu lie Asp Tyr Gly Asp Gin Asp lie 95 100 105 ATC ACT TTA ATG ATC ATT AAT AAG TTA CTA TCG ATA GAT GAT ATA TCC 5 lie Thr Leu Met lie lie Asn Lys Leu Leu Ser lie Asp Asp lie Ser 110 115 120 TAT ATA TTA GAT AAA AAA ATA ATT CAT GTA AC 5 Tyr lie Leu Asp Lys Lys lie lie His Val 125 130 (2) INFORMATION FOR SEQ ID NO:2: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 132 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2: Met Pro Ser Tyr Met Tyr Pro Lys Asn Ala Arg Lys Val lie Ser Lys 1 5 10 15 lie lie Ser Leu Gin Leu Asp lie Lys Lys Leu Pro Lys Lys Tyr lie 20 25 30 Asn Thr Met Leu Glu Phe Gly Leu His Gly Asn Leu Pro Ala Cys Met 35 40 45 Tyr Lys Asp Ala Val Ser Tyr Asp lie Asn Asn lie Arg Phe Leu Pro 50 55 60 Tyr Asn Cys Val Met Val Lys Asp Leu lie Asn Val lie Lys Ser Ser 65 70 75 80 Ser Val lie Asp Thr Arg Leu His Gin Ser Val Leu Lys His Arg Arg 85 90 95 Ala Leu lie Asp Tyr Gly Asp Gin Asp lie lie Thr Leu Met lie lie 100 105 110 Asn Lys Leu Leu Ser lie Asp Asp lie Ser Tyr lie Leu Asp Lys Lys 115 120 125 lie lie His Val 130 (2) INFORMATION FOR SEQ ID NO:3: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 899 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (vi) ORIGINAL SOURCE: (A) ORGANISM: Swinepox virus (B) STRAIN: Kasza (C) INDIVIDUAL ISOLATE: S-SPV-001 (vii) IMMEDIATE SOURCE: (B) CLONE: 515-85.1 (viii) POSITION IN GENOME: (B) MAP POSITION: -23.2 (C) UNITS: %G (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION". 3..662 (D) OTHER INFORMATION: /partial /codon_εtart= 3 /function= "Potential eukaryotic transcriptional regulatory protein" /standard name= "515-85.1 ORF" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3: GA GAT ATT AAA TCA TGT AAA TGC TCG ATA TGT TCC GAC TCT ATA ACA Asp lie Lys Ser Cys Lys Cys Ser lie Cys Ser Asp Ser lie Thr 1 5 10 15 CAT CAT ATA TAT GAA ACA ACA TCA TGT ATA AAT TAT AAA TCT ACC GAT His His lie Tyr Glu Thr Thr Ser Cys lie Asn Tyr Lys Ser Thr Asp 20 25 30 AAT GAT CTT ATG ATA GTA TTG TTC AAT CTA ACT AGA TAT TTA ATG CAT 1 Asn Asp Leu Met lie Val Leu Phe Asn Leu Thr Arg Tyr Leu Met His 35 40 45 GGG ATG ATA CAT CCT AAT CTT ATA AGC GTA AAA GGA TGG GGT CCC CTT 1 Gly Met lie His Pro Asn Leu lie Ser Val Lys Gly Trp Gly Pro Leu 50 55 60 ATT GGA TTA TTA ACG GGT GAT ATA GGT ATT AAT TTA AAA CTA TAT TCC 2 lie Gly Leu Leu Thr Gly Asp lie Gly He Asn Leu Lys Leu Tyr Ser 65 70 75 ACC ATG AAT ATA AAT GGG CTA CGG TAT GGA GAT ATT ACG TTA TCT TCA 2 Thr Met Asn He Asn Gly Leu Arg Tyr Gly Asp He Thr Leu Ser Ser 80 85 90 95 TAC GAT ATG AGT AAT AAA TTA GTC TCT ATT ATT AAT ACA CCC ATA TAT 3 Tyr Asp Met Ser Asn Lys Leu Val Ser He He Asn Thr Pro He Tyr 100 105 110 GAG TTA ATA CCG TTT ACT ACA TGT TGT TCA CTC AAT GAA TAT TAT TCA 3 Glu Leu He Pro Phe Thr Thr Cys Cys Ser Leu Asn Glu Tyr Tyr Ser 115 120 125 AAA ATT GTG ATT TTA ATA AAT GTT ATT TTA GAA TAT ATG ATA TCT ATT 4 Lys He Val He Leu He Asn Val He Leu Glu Tyr Met He Ser He 130 135 140 ATA TTA TAT AGA ATA TTG ATC GTA AAA AGA TTT AAT AAC ATT AAA GAA 4 He Leu Tyr Arg He Leu He Val Lys Arg Phe Asn Asn He Lys Glu 145 150 155 TTT ATT TCA AAA GTC GTA AAT ACT GTA CTA GAA TCA TCA GGC ATA TAT 5 Phe He Ser Lys Val Val Asn Thr Val Leu Glu Ser Ser Gly He Tyr 160 165 170 175 TTT TGT CAG ATG CGT GTA CAT GAA CAA ATT GAA TTG GAA ATA GAT GAG 5 Phe Cys Gin Met Arg Val His Glu Gin He Glu Leu Glu He Asp Glu 180 185 190 CTC ATT ATT AAT GGA TCT ATG CCT GTA CAG CTT ATG CAT TTA CTT CTA 6 Leu He He Asn Gly Ser Met Pro Val Gin Leu Met His Leu Leu Leu 195 200 205 AAG GTA GCT ACC ATA ATA TTA GAG GAA ATC AAA GAA ATA TAACGTATTT 6 Lys Val Ala Thr He He Leu Glu Glu He Lys Glu He 210 215 220 TTTCTTTTAA ATAAATAAAA ATACTTTTTT TTTTAAACAA GGGGTGCTAC CTTGTCTAAT 7 TGTATCTTGT ATTTTGGATC TGATGCAAGA TTATTAAATA ATCGTATGAA AAAGTAGTAG 7 ATATAGTTTA TATCGTTACT GGACATGATA TTATGTTTAG TTAATTCTTC TTTGGCATGA 8 ATTCTACACG TCGGANAAGG TAATGTATCT ATAATGGTAT AAAGCTT 8 (2) INFORMATION FOR SEQ ID NO:4: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 220 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:4: Asp He Lys Ser Cys Lys Cys Ser He Cys Ser Asp Ser He Thr His 1 5 10 15 His He Tyr Glu Thr Thr Ser Cys He Asn Tyr Lys Ser Thr Asp Asn 20 25 30 Asp Leu Met He Val Leu Phe Asn Leu Thr Arg Tyr Leu Met His Gly 35 40 45 Met He His Pro Asn Leu He Ser Val Lys Gly Trp Gly Pro Leu He 50 55 60 Gly Leu Leu Thr Gly Asp He Gly He Asn Leu Lys Leu Tyr Ser Thr 65 70 75 80 Met Asn He Asn Gly Leu Arg Tyr Gly Asp He Thr Leu Ser Ser Tyr 85 90 95 Asp Met Ser Asn Lys Leu Val Ser He He Asn Thr Pro He Tyr Glu 100 105 110 Leu He Pro Phe Thr Thr Cys Cys Ser Leu Asn Glu Tyr Tyr Ser Lys 115 120 125 He Val He Leu He Asn Val He Leu Glu Tyr Met He Ser He He 130 135 140 Leu Tyr Arg He Leu He Val Lys Arg Phe Asn Asn He Lys Glu Phe 145 150 155 160 He Ser Lys Val Val Asn Thr Val Leu Glu Ser Ser Gly He Tyr Phe 165 170 175 Cys Gin Met Arg Val His Glu Gin He Glu Leu Glu He Asp Glu Leu 180 185 190 He He Asn Gly Ser Met Pro Val Gin Leu Met His Leu Leu Leu Lys 195 200 205 Val Ala Thr He He Leu Glu Glu He Lys Glu He 210 215 220 (2) INFORMATION FOR SEQ ID NO:5: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 129 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: YES (iv) ANTI-SENSE: NO (v) FRAGMENT TYPE: N-terminal (vi) ORIGINAL SOURCE: (A) ORGANISM: Vaccinia virus (B) STRAIN: Copenhagen (viii) POSITION IN GENOME: (B) MAP POSITION: -23. (C) UNITS: %G (xi ) SEQUENCE DESCRIPTION : SEQ ID NO: 5 : Met Phe Met Tyr Pro Glu Phe Ala Arg Lys Ala Leu Ser Lys Leu He 1 5 10 15 Ser Lys Lys Leu Asn He Glu Lys Val Ser Ser Lys His Gin Leu Val 20 25 30 Leu Leu Asp Tyr Gly Leu His Gly Leu Leu Pro Lys Ser Leu Tyr Leu 35 40 45 Glu Ala He Asn Ser Asp He Leu Asn Val Arg Phe Phe Pro Pro Glu 50 55 60 He He Asn Val Thr Asp He Val Lys Ala Leu Gin Asn Ser Cys Arg 65 70 75 80 Val Asp Glu Tyr Leu Lys Ala Val Ser Leu Tyr His Lys Asn Ser Leu 85 90 95 Met Val Ser Gly Pro Asn Val Val Lys Leu Met He Glu Tyr Asn Leu 100 105 110 Leu Thr His Ser Asp Leu Glu Trp Leu He Asn Glu Asn Val Val Lys 115 120 125 Ala (2) INFORMATION FOR SEQ ID NO:6: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 132 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: YES (iv) ANTI-SENSE: NO (v) FRAGMENT TYPE: N-terminal (vi) ORIGINAL SOURCE: (A) ORGANISM: Swinepox virus (B) STRAIN: Kasza (viii) POSITION IN GENOME: (B) MAP POSITION: -23.2 (C) UNITS: %G (xi) SEQUENCE DESCRIPTION: SEQ ID NO:6: Met Pro Ser Tyr Met Tyr Pro Lys Asn Ala Arg Lys Val He Ser Lys 1 5 10 15 He He Ser Leu Gin Leu Asp He Lys Lys Leu Pro Lys Lys Tyr He 20 25 30 Asn Thr Met Leu Glu Phe Gly Leu His Gly Asn Leu Pro Ala Cys Met 35 40 45 Tyr Lys Asp Ala Val Ser Tyr Asp He Asn Asn He Arg Phe Leu Pro 50 55 60 Tyr Asn Cys Val Met Val Lys Asp Leu He Asn Val He Lys Ser Ser 65 70 75 80 Ser Val He Asp Thr Arg Leu His Gin Ser Val Leu Lys His Arg Arg 85 90 95 Ala Leu He Asp Tyr Gly Asp Gin Asp He He Thr Leu Met He He 100 105 110 Asn Lys Leu Leu Ser He Asp Asp He Ser Tyr He Leu Asp Lys Lys 115 120 125 He He His Val 130 (2) INFORMATION FOR SEQ ID NO:7: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 101 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: YES (iv) ANTI-SENSE: NO (v) FRAGMENT TYPE: C-terminal (vi) ORIGINAL SOURCE: (A) ORGANISM: Vaccinia virus (B) STRAIN: Copenhagen (viii) POSITION IN GENOME: (B) MAP POSITION: -23.2 (C) UNITS: %G (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 1 : Val Leu Asn Asp Gin Tyr Ala Lys He Val He Phe Phe Asn Thr He 1 5 10 15 He Glu Tyr He He Ala Thr He Tyr Tyr Arg Leu Thr Val Leu Asn 20 25 30 Asn Tyr Thr Asn Val Lys His Phe Val Ser Lys Val Leu His Thr Val 35 40 45 Met Glu Ala Cys Gly Val Leu Phe Ser Tyr He Lys Val Asn Asp Lys 50 55 60 He Glu His Glu Leu Glu Glu Met Val Asp Lys Gly Thr Val Pro Ser 65 70 75 80 Tyr Leu Tyr His Leu Ser He Asn Val He Ser He He Leu Asp Asp 85 90 95 He Asn Gly Thr Arg 100 (2) INFORMATION FOR SEQ ID NO:8: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 100 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: YES (iv) ANTI-SENSE: NO (v) FRAGMENT TYPE: C-terminal (vi) ORIGINAL SOURCE: (A) ORGANISM: Swinepox virus (B) STRAIN: Kasza (viii) POSITION IN GENOME: (B) MAP POSITION: -23.2 (C) UNITS: %G ( xi ) SEQUENCE DESCRIPTION : SEQ ID NO: 8 : Ser Leu Asn Glu Tyr Tyr Ser Lys He Val He Leu He Asn Val He 1 5 10 15 Leu Glu Tyr Met He Ser He He Leu Tyr Arg He Leu He Val Lys 20 25 30 Arg Phe Asn Asn He Lys Glu Phe He Ser Lys Val Val Asn Thr Val 35 40 45 Leu Glu Ser Ser Gly He Tyr Phe Cys Gin Met Arg Val His Glu Gin 50 55 60 He Glu Leu Glu He Asp Glu Leu He He Asn Gly Ser Met Pro Val 65 70 75 80 Gin Leu Met His Leu Leu Leu Lys Val Ala Thr He He Leu Glu Glu 85 90 95 He Lys Glu He 100 (2) INFORMATION FOR SEQ ID NO:9: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 102 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: circular (ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (vi) ORIGINAL SOURCE: (A) ORGANISM: Plasmid (vii) IMMEDIATE SOURCE: (B) CLONE: 520-17.5 (Junction A) (x) PUBLICATION INFORMATION: (A) AUTHORS: Ferrari, Franco A Trach, Kathleen Hoch, James A (B) TITLE: Sequence Analysis of the spoOB Locus Revels a Polycistronic Transcription Unit (C) JOURNAL: J. Bacteriol. (D) VOLUME: 161 (E) ISSUE: 2 (F) PAGES: 556-562 (G) DATE: Feb.-1985 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:9: CACATACGAT TTAGGTGACA CTATAGAATA CAAGCTTTAT ACCATTATAG ATACATTACC 6 TTGTCCGACG TGTAGAATTC ATGCCAAAGA AGAATTAACT AA 10 (2) INFORMATION FOR SEQ ID NO:10: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 102 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: circular (ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (vi) ORIGINAL SOURCE: (A) ORGANISM: Plasmid (vii) IMMEDIATE SOURCE: (B) CLONE: 520-17.5 (Junction B) (ix) FEATURE: , ._ ,_, ,_._-. TITUTE SHEET (A) NAME/KEY: CDS (B) LOCATION: 85..99 (D) OTHER INFORMATION: /codon_start= 85 /function= "Translational start of hybrid protein" /product= "N-terminal peptide" /number= 1 /standard_name= "Translation of synthetic DNA sequence" (i ) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 100..102 (C) IDENTIFICATION METHOD: experimental (D) OTHER INFORMATION: /partial /codon_start= 100 /function= "marker enzyme" /product= "Beta-Galactosidase" /evidence * -- * EXPERIMENTAL /gene= "lacZ" /number= 2 /citation= ([1]) (x) PUBLICATION INFORMATION: (A) AUTHORS: Ferrari, Franco A Trach, Kathleen Hoch, James A (B) TITLE: Seqquence Analysis of the spoOB Locus Reveals a Polycistronic Transcription Unit (C) JOURNAL: J. Bacteriol. (D) VOLUME: 161 (E) ISSUE: 2 (F) PAGES: 556-562 (G) DATE: Feb.-1985 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 10: GTAGTCGACT CTAGAAAAAA TTGAAAAACT ATTCTAATTT ATTGCACGGA GATCTTTTTT 6 TTTTTTTTTT TTTTTGGCAT ATAA ATG AAT TCG GAT CCC GTC 10 Met Asn Ser Asp Pro Val 1 5 1 (2) INFORMATION FOR SEQ ID NO:11: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 5 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:ll: Met Asn Ser Asp Pro 1 5 (2) INFORMATION FOR SEQ ID NO: 12: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 1 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear ET (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:12: Val 1 (2) INFORMATION FOR SEQ ID NO:13: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 103 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: circular (ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (vi) ORIGINAL SOURCE: (A) ORGANISM: Plasmid (vii) IMMEDIATE SOURCE: (B) CLONE: 520-17.5 (Junction C) (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 1..72 (C) IDENTIFICATION METHOD: experimental (D) OTHER INFORMATION: /partial /codon_start= 1 /function-- * - "marker enzyme" /product= "Beta-galactosidase" /evidence= EXPERIMENTAL /gene= "lacZ" /numbe = 1 /citation= ([1]) (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 73..78 (C) IDENTIFICATION METHOD: experimental (D) OTHER INFORMATION: /codon_start= 73 /function= "Translational finish of hybrid protein" /product= "C-terminal peptide" /evidence= EXPERIMENTAL /number= 2 /standard_name= "Translation of synthetic DNA sequence" (x) PUBLICATION INFORMATION: (A) AUTHORS: Ferrari, Franco A Trach, Kathleen Hoch, James A (B) TITLE: Seqquence Analysis of the spoOB Locus Reveals a Polycistronic Transcription Unit (C) JOURNAL: J. Bacteriol. (D) VOLUME: 161 (E) ISSUE: 2 (F) PAGES: 556-562 (G) DATE: Feb.-1985 ( i) SEQUENCE DESCRIPTION: SEQ ID NO: 13: AGC CCG TCA GTA TCG GCG GAA ATC CAG CTG AGC GCC GGT CGC TAC CAT Ser Pro Ser Val Ser Ala Glu He Gin Leu Ser Ala Gly Arg Tyr His 1 5 10 15 TAC CAG TTG GTC TGG TGT CAA AAA GAT CCA TAATTAATTA ACCCGGGTCG Tyr Gin Leu Val Trp Cys Gin Lys Asp Pro 20 1 AAGAC 1 (2) INFORMATION FOR SEQ ID NO:14: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 24 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein ( i) SEQUENCE DESCRIPTION: SEQ ID NO:14: Ser Pro Ser Val Ser Ala Glu He Gin Leu Ser Ala Gly Arg Tyr His 1 5 10 15 Tyr Gin Leu Val Trp Cys Gin Lys 20 (2) INFORMATION FOR SEQ ID NO: 15: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 2 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein ( i) SEQUENCE DESCRIPTION: SEQ ID NO: 15: sp Pro 1 (2) INFORMATION FOR SEQ ID NO: 16: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 48 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: circular (ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (vi) ORIGINAL SOURCE: (A) ORGANISM: Plasmid (vii) IMMEDIATE SOURCE: (B) CLONE: 520-17.5 (Junction D) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:16: AGATCCCCGG GCGAGCTCGA ATTCGTAATC ATGGTCATAG CTGTTTCC 4 (2) INFORMATION FOR SEQ ID NO:17: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 57 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: circular (ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (vi) ORIGINAL SOURCE: (A) ORGANISM: Plasmid (vii) IMMEDIATE SOURCE: (B) CLONE: 538-46.26 (Junction A) (Xi) SEQUENCE DESCRIPTION: SEQ ID NO:17: CACATACGAT TTAGGTGACA CTATAGAATA CAAGCTTTAT ACCATTATAG ATACATT 57 (2) INFORMATION FOR SEQ ID NO:18: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 102 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: circular (ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (vi) ORIGINAL SOURCE: (A) ORGANISM: Plasmid (vii) IMMEDIATE SOURCE: (B) CLONE: 538-46.16 (Junction B) (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 91..102 (C) IDENTIFICATION METHOD: experimental (D) OTHER INFORMATION: /partial /codon_start= 91 /function= "marker enzyme" /product= "Beta-Galactosidase" /evidence= EXPERIMENTAL /gene= "lacZ" /number= 2 /citation= ([1]) (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 76..90 (D) OTHER INFORMATION: /partial /codon_start= 76 /function= "Translational start of hybrid protein" /product= "N-terminal peptide" /number= 1 /standard_name= "Translation of synthetic DNA sequence" (X) PUBLICATION INFORMATION: (A) AUTHORS: Ferrari, Franco A Trach, Kathleen Hoch, James A (B) TITLE: Seqquence Analysis of the spoOB Locus Reveals a Polycistronic Transcription Unit (C) JOURNAL: J. Bacteriol. (D) VOLUME: 161 (E) ISSUE: 2 (F) PAGES: 556-562 (G) DATE: Feb.-1985 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 18: AAGCTGGTAG ATTTCCATGT AGGGCCGCCT GCAGGTCGAC TCTAGAATTT CATTTTGTTT TTTTCTATGC TATAA ATG AAT TCG GAT CCC GTC GTT TTA CAA 1 Met Asn Ser Asp Pro Val Val Leu Gin 1 5 1 (2) INFORMATION FOR SEQ ID NO:19: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 5 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 19: Met Asn Ser Asp Pro 1 5 (2) INFORMATION FOR SEQ ID NO:20: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:20: Val Val Leu Gin 1 (2) INFORMATION FOR SEQ ID NO:21: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 206 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: circular (ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (vi) ORIGINAL SOURCE: (A) ORGANISM: Plasmid (vii) IMMEDIATE SOURCE: (B) CLONE: 538-46.16 (Junction C) (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 1..63 (C) IDENTIFICATION METHOD: experimental (D) OTHER INFORMATION: /partial /codon_s art= 1 /function= "marker enzyme" /product= "Beta-galactosidase" /evidences EXPERIMENTAL /number= 1 /citations ([1]) (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 64..69 (C) IDENTIFICATION METHOD: experimental (D) OTHER INFORMATION: /codon_start= 64 /functions "Translational finish of hybrid protein" /products "C-terminal peptide" /evidences EXPERIMENTAL /standard_name= "Translation of synthetic DNA sequence" (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 177..185 (C) (D) ein" (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 186..206 (C) IDENTIFICATION METHOD: experimental (D) OTHER INFORMATION: /partial /codon_start = 186 /functions "glycoprotein" /products "PRV gp50" /evidences EXPERIMENTAL /gene= "gp50" /numbers 3 /citations ([2]) (X) PUBLICATION INFORMATION: (A) AUTHORS: Ferrari, Franco A Trach, Kathleen Hoch, James A (B) TITLE: Seqquence Analysis of the spoOB Locus Reveals a Polyciεtronic Transcription Unit (C) JOURNAL: J. Bacteriol. (D) VOLUME: 161 (E) ISSUE: 2 (F) PAGES: 556-562 (G) DATE: Feb.-1985 (X) PUBLICATION INFORMATION: (A) AUTHORS: Petrovskis, Erik A Timmins, James G Armentrout, Marty A Marchioli, Carmine C Jr. Yancy, Robert J Post, Leonard E (B) TITLE: DNA Sequence of the Gene for Pseudorabies Virus gp50, a Glycoprotein without N-Linked Glycosylation (C) JOURNAL: J. Virol. (D) VOLUME: 59 (E) ISSUE: 2 (F) PAGES: 216-223 (G) DATE: Aug.-1986 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:21: GTA TCG GCG GAA ATC CAG CTG AGC GCC GGT CGC TAC CAT TAC CAG TTG Val Ser Ala Glu He Gin Leu Ser Ala Gly Arg Tyr His Tyr Gin Leu 1 5 10 15 GTC TGG TGT CAA AAA GAT CCA TAATTAATTA ACCCGGCCGC CTGCAGGTCG Val Trp Cys Gin Lys Asp Pro 20 1 ACTCTAGAAA AAATTGAAAA ACTATTCTAA TTTATTGCAC GGAGATCTTT TTTTTTTTTT 1 TTTTTTTTGG CATATAA ATG AAT TCG CTC GCA GCG CTA TTG GCG GCG 2 Met Asn Ser Leu Ala Ala Leu Leu Ala Ala 1 1 5 (2) INFORMATION FOR SEQ ID NO:22: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 21 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:22: Val Ser Ala Glu He Gin Leu Ser Ala Gly Arg Tyr His Tyr Gin Leu 1 5 10 15 Val Trp Cys Gin Lys 20 (2) INFORMATION FOR SEQ ID NO:23: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 2 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:23: Asp Pro 1 (2) INFORMATION FOR SEQ ID NO:24: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 3 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:24: Met Asn Ser 1 (2) INFORMATION FOR SEQ ID NO:25: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 7 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein ( i) SEQUENCE DESCRIPTION: SEQ ID NO:25: Leu Ala Ala Leu Leu Ala Ala 1 5 (2) INFORMATION FOR SEQ ID NO:26: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 101 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: circular (ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (vi) ORIGINAL SOURCE: (A) ORGANISM: Plasmid (vii) IMMEDIATE SOURCE: (B) CLONE: 538-46.16 (Junction D) (ix) FEATUREs (A) NAME/KEY: CDS (B) LOCATION: 1..15 (D) OTHER INFORMATION: /partial /codon_starts l /functions "glycoprotein" /products "PRV gp63" /gene = "gp63" /numbers i /citations ([l]) (x) PUBLICATION INFORMATION: (A) AUTHORS: Petrovskis, Erik A Timminε, James G Post, Lenoard E (B) TITLE: Use of Lambda-gtll To Isolate Genes for two Pseudorabies Virus Glycoproteins with homology to Herpes Simplex Virus and Varicella-Zoster Virus Glycoproteins (C) JOURNAL: J. Virol. (D) VOLUME: 60 (E) ISSUE: 1 (F) PAGES: 185-193 (G) DATE: Oct.-1986 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:26: CGC GTG CAC CAC GAG GGACTCTAGA GGATCCATAA TTAATTAATT AATTTTTATC Arg Val His His Glu 1 5 CCGGGTCGAC CTGCAGGCGG CCGGGTCGAC CTGCAGGCGG CCAGAC 1 (2) INFORMATION FOR SEQ ID NO:27: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 5 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:27: Arg Val His His Glu 1 5 (2) INFORMATION FOR SEQ ID NO:28: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 57 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: circular (ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (vi) ORIGINAL SOURCE: (A) ORGANISM: Plasmid (vii) IMMEDIATE SOURCE: (B) CLONE: 538-46.16 (Junction E) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:28: AGATCCCCGG GCGAGCTCGA ATTCGTAATC ATGGTCATAG CTGTTTCCTG TGTGAAA 5 (2) INFORMATION FOR SEQ ID NO:29: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 1907 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: cDNA to mRNA (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (vi) ORIGINAL SOURCE: (A) ORGANISM: Newcastle disease virus (B) STRAIN: Bl (vii) IMMEDIATE SOURCE: (B) CLONE: 137-23.803 (PSY1142) (viii) POSITION IN GENOME: (B) MAP POSITION: -50% (C) UNITS: %G (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 92..1822 (D) OTHER INFORMATION: /codon_start= 92 /products "NDV heamagglutinin-Neuraminidase" /genes » HN" /numbers l (xi) SEQUENCE DESCRIPTION: SEQ ID NO:29: ACGGGTAGAA CGGTAAGAGA GGCCGCCCCT CAATTGCGAG CCAGACTTCA CAACCTCCGT 60 TCTACCGCTT CACCGACAAC AGTCCTCAAT C ATG GAC CGC GCC GTT AGC CAA 112 Met Asp Arg Ala Val Ser Gin 1 5 GTT GCG TTA GAG AAT GAT GAA AGA GAG GCA AAA AAT ACA TGG CGC TTG 160 Val Ala Leu Glu Asn Asp Glu Arg Glu Ala Lys Asn Thr Trp Arg Leu 10 ' 15 20 ATA TTC CGG ATT GCA ATC TTA TTC TTA ACA GTA GTG ACC TTG GCT ATA 208 He Phe Arg He Ala He Leu Phe Leu Thr Val Val Thr Leu Ala He 25 30 35 TCT GTA GCC TCC CTT TTA TAT AGC ATG GGG GCT AGC ACA CCT AGC GAT 256 Ser Val Ala Ser Leu Leu Tyr Ser Met Gly Ala Ser Thr Pro Ser Asp 40 45 50 55 CTT GTA GGC ATA CCG ACT AGG ATT TCC AGG GCA GAA GAA AAG ATT ACA 304 Leu Val Gly He Pro Thr Arg He Ser Arg Ala Glu Glu Lys He Thr 60 65 70 TCT ACA CTT GGT TCC AAT CAA GAT GTA GTA GAT AGG ATA TAT AAG CAA 352 Ser Thr Leu Gly Ser Asn Gin Asp Val Val Asp Arg He Tvr Lys Gin 75 80 85 GTG GCC CTT GAG TCT CCA TTG GCA TTG TTA AAT ACT GAG ACC ACA ATT 400 Val Ala Leu Glu Ser Pro Leu Ala Leu Leu Asn Thr Glu Thr Thr He 90 95 100 ATG AAC GCA ATA ACA TCT CTC TCT TAT CAG ATT AAT GGA GCT GCA AAC Met Asn Ala He Thr Ser Leu Ser Tyr Gin He Asn Gly Ala Ala Asn 105 110 115 AAC AGC GGG TGG GGG GCA CCT ATT CAT GAC CCA GAT TAT ATA GGG GGG Asn Ser Gly Trp Gly Ala Pro He His Asp Pro Asp Tyr He Gly Gly 120 125 130 135 ATA GGC AAA GAA CTC ATT GTA GAT GAT GCT AGT GAT GTC ACA TCA TTC He Gly Lys Glu Leu He Val Asp Asp Ala Ser Asp Val Thr Ser Phe 140 145 150 TAT CCC TCT GCA TTT CAA GAA CAT CTG AAT TTT ATC CCG GCG CCT ACT Tyr Pro Ser Ala Phe Gin Glu His Leu Asn Phe He Pro Ala Pro Thr 155 160 165 ACA GGA TCA GGT TGC ACT CGA ATA CCC TCA TTT GAC ATG AGT ' GCT ACC Thr Gly Ser Gly Cys Thr Arg He Pro Ser Phe Asp Met Ser Ala Thr 170 175 180 CAT TAC TGC TAC ACC CAT AAT GTA ATA TTG TCT GGA TGC AGA GAT CAC His Tyr Cys Tyr Thr His Asn Val He Leu Ser Gly Cys Arg Asp His 185 190 195 TCA CAC TCA CAT CAG TAT TTA GCA CTT GGT GTG CTC CGG ACA TCT GCA Ser His Ser His Gin Tyr Leu Ala Leu Gly Val Leu Arg Thr Ser Ala 200 205 210 215 ACA GGG AGG GTA TTC TTT TCT ACT CTG CGT TCC ATC AAC CTG GAC GAC Thr Gly Arg Val Phe Phe Ser Thr Leu Arg Ser He Asn Leu Asp Asp 220 225 230 ACC CAA AAT CGG AAG TCT TGC AGT GTG AGT GCA ACT CCC CTG GGT TGT Thr Gin Asn Arg Lys Ser Cys Ser Val Ser Ala Thr Pro Leu Gly Cys 235 240 245 GAT ATG CTG TGC TCG AAA GCC ACG GAG ACA GAG GAA GAA GAT TAT AAC Asp Met Leu Cys Ser Lys Ala Thr Glu Thr Glu Glu Glu Asp Tyr Asn 250 255 260 TCA GCT GTC CCT ACG CGG ATG GTA CAT GGG AGG TTA GGG TTC GAC GGC Ser Ala Val Pro Thr Arg Met Val His Gly Arg Leu Gly Phe Asp Gly 265 270 275 CAA TAT CAC GAA AAG GAC CTA GAT GTC ACA ACA TTA TTC GGG GAC TGG 9 Gin Tyr His Glu Lys Asp Leu Asp Val Thr Thr Leu Phe Gly Asp Trp 280 285 290 295 GTG GCC AAC TAC CCA GGA GTA GGG GGT GGA TCT TTT ATT GAC AGC CGC 10 Val Ala Asn Tyr Pro Gly Val Gly Gly Gly Ser Phe He Asp Ser Arg 300 305 310 GTG TGG TTC TCA GTC TAC GGA GGG TTA AAA CCC AAT ACA CCC AGT GAC 10 Val Trp Phe Ser Val Tyr Gly Gly Leu Lys Pro Asn Thr Pro Ser Asp 315 320 325 ACT GTA CAG GAA GGG AAA TAT GTG ATA TAC AAG CGA TAC AAT GAC ACA 11 Thr Val Gin Glu Gly Lys Tyr Val He Tyr Lys Arg Tyr Asn Asp Thr 330 335 340 TGC CCA GAT GAG CAA GAC TAC CAG ATT CGA ATG GCC AAG TCT TCG TAT 11 Cys Pro Asp Glu Gin Asp Tyr Gin He Arg Met Ala Lys Ser Ser Tyr 345 350 355 AAG CCT GGA CGG TTT GGT GGG AAA CGC ATA CAG CAG GCT ATC TTA TCT 1216 Lys Pro Gly Arg Phe Gly Gly Lys Arg He Gin Gin Ala He Leu Ser 360 365 370 375 ATC AAA GTG TCA ACA TCC TTA GGC GAA GAC CCG GTA CTG ACT GTA CCG 1264 He Lys Val Ser Thr Ser Leu Gly Glu Asp Pro Val Leu Thr Val Pro 380 385 390 CCC AAC ACA GTC ACA CTC ATG GGG GCC GAA GGC AGA ATT CTC ACA GTA 1312 Pro Asn Thr Val Thr Leu Met Gly Ala Glu Gly Arg He Leu Thr Val 395 400 405 GGG ACA TCC CAT TTC TTG TAT CAG CGA GGG TCA TCA TAC TTC TCT CCC 1360 Gly Thr Ser His Phe Leu Tyr Gin Arg Gly Ser Ser Tyr Phe Ser Pro 410 415 420 GCG TTA TTA TAT CCT ATG ACA GTC AGC AAC AAA ACA GCC ACT CTT CAT 1408 Ala Leu Leu Tyr Pro Met Thr Val Ser Asn Lys Thr Ala Thr Leu His 425 430 435 AGT CCT TAT ACA TTC AAT GCC TTC ACT CGG CCA GGT AGT ATC CCT TGC 1456 Ser Pro Tyr Thr Phe Asn Ala Phe Thr Arg Pro Gly Ser He Pro Cys 440 445 450 455 CAG GCT TCA GCA AGA TGC CCC AAC TCA TGT GTT ACT GGA GTC TAT ACA 1504 Gin Ala Ser Ala Arg Cys Pro Asn Ser Cys Val Thr Gly Val Tyr Thr 460 465 470 GAT CCA TAT CCC CTA ATC TTC TAT AGA AAC CAC ACC TTG CGA GGG GTA 1552 Asp Pro Tyr Pro Leu He Phe Tyr Arg Asn His Thr Leu Arg Gly Val 475 480 485 TTC GGG ACA ATG CTT GAT GGT GAA CAA GCA AGA CTT AAC CCT GCG TCT 1600 Phe Gly Thr Met Leu Asp Gly Glu Gin Ala Arg Leu Asn Pro Ala Ser 490 495 500 GCA GTA TTC GAT AGC ACA TCC CGC AGT CGC ATA ACT CGA GTG AGT TCA 1648 Ala Val Phe Asp Ser Thr Ser Arg Ser Arg He Thr Arg Val Ser Ser 505 510 515 AGC AGC ATC AAA GCA GCA TAC ACA ACA TCA ACT TGT TTT AAA GTG GTC 1696 Ser Ser He Lys Ala Ala Tyr Thr Thr Ser Thr Cys Phe Lys Val Val 520 525 530 535 AAG ACC AAT AAG ACC TAT TGT CTC AGC ATT GCT GAA ATA TCT AAT ACT 1744 Lys Thr Asn Lys Thr Tyr Cys Leu Ser He Ala Glu He Ser Asn Thr 540 545 550 CTC TTC GGA GAA TTC AGA ATC GTC CCG TTA CTA GTT GAG ATC CTC AAA 1792 Leu Phe Gly Glu Phe Arg He Val Pro Leu Leu Val Glu He Leu Lys 555 560 565 GAT GAC GGG GTT AGA GAA GCC AGG TCT GGC TAGTTGAGTC AACTATGAAA 1842 Asp Asp Gly Val Arg Glu Ala Arg Ser Gly 570 575 GAGTTGGAAA GATGGCATTG TATCACCTAT CTTCTGCGAC ATCAAGAATC AAACCGAATG 1902 CCGGC 1907 (2) INFORMATION FOR SEQ ID NO:30: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 577 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:30: Met Asp Arg Ala Val Ser Gin Val Ala Leu Glu Asn Asp Glu Arg Glu 1 5 10 15 Ala Lys Asn Thr Trp Arg Leu He Phe Arg He Ala He Leu Phe Leu 20 25 30 Thr Val Val Thr Leu Ala He Ser Val Ala Ser Leu Leu Tyr Ser Met 35 40 45 Gly Ala Ser Thr Pro Ser Asp Leu Val Gly He Pro Thr Arg He Ser 50 55 60 Arg Ala Glu Glu Lys He Thr Ser Thr Leu Gly Ser Asn Gin Asp Val 65 70 75 80 Val Asp Arg He Tyr Lys Gin Val Ala Leu Glu Ser Pro Leu Ala Leu 85 90 95 Leu Asn Thr Glu Thr Thr He Met Asn Ala He Thr Ser Leu Ser Tyr 100 105 110 Gin He Asn Gly Ala Ala Asn Asn Ser Gly Trp Gly Ala Pro He His 115 120 125 Asp Pro Asp Tyr He Gly Gly He Gly Lys Glu Leu He Val Asp Asp 130 135 140 Ala Ser Asp Val Thr Ser Phe Tyr Pro Ser Ala Phe Gin Glu His Leu 145 150 155 160 Asn Phe He Pro Ala Pro Thr Thr Gly Ser Gly Cys Thr Arg He Pro 165 170 175 Ser Phe Asp Met Ser Ala Thr His Tyr Cys Tyr Thr His Asn Val He 180 185 190 Leu Ser Gly Cys Arg Asp His Ser His Ser His Gin Tyr Leu Ala Leu 195 200 205 Gly Val Leu Arg Thr Ser Ala Thr Gly Arg Val Phe Phe Ser Thr Leu 210 215 220 Arg Ser He Asn Leu Asp Asp Thr Gin Asn Arg Lys Ser Cys Ser Val 225 230 235 240 Ser Ala Thr Pro Leu Gly Cys Asp Met Leu Cys Ser Lys Ala Thr Glu 245 250 255 Thr Glu Glu Glu Asp Tyr Asn Ser Ala Val Pro Thr Arg Met Val His 260 265 270 Gly Arg Leu Gly Phe Asp Gly Gin Tyr His Glu Lys Asp Leu Asp Val 275 280 285 Thr Thr Leu Phe Gly Asp Trp Val Ala Asn Tyr Pro Gly Val Gly Gly 290 295 300 Gly Ser Phe He Asp Ser Arg Val Trp Phe Ser Val Tyr Gly Gly Leu 305 310 315 320 Lys Pro Asn Thr Pro Ser Asp Thr Val Gin Glu Gly Lys Tyr Val He 325 330 335 Tyr Lys Arg Tyr Asn Asp Thr Cys Pro Asp Glu Gin Asp Tyr Gin He 340 345 350 Arg Met Ala Lys Ser Ser Tyr Lys Pro Gly Arg Phe Gly Gly Lys Arg 355 360 365 He Gin Gin Ala He Leu Ser He Lys Val Ser Thr Ser Leu Gly Glu 370 375 380 Asp Pro Val Leu Thr Val Pro Pro Asn Thr Val Thr Leu Met Gly Ala 385 390 395 400 Glu Gly Arg He Leu Thr Val Gly Thr Ser His Phe Leu Tyr Gin Arg 405 410 415 Gly Ser Ser Tyr Phe Ser Pro Ala Leu Leu Tyr Pro Met Thr Val Ser 420 425 430 Asn Lys Thr Ala Thr Leu His Ser Pro Tyr Thr Phe Asn Ala Phe Thr 435 440 445 Arg Pro Gly Ser He Pro Cys Gin Ala Ser Ala Arg Cys Pro Asn Ser 450 455 460 Cvs Val Thr Gly Val Tyr Thr Asp Pro Tyr Pro Leu He Phe Tyr Arg 465 470 475 480 Asn His Thr Leu Arg Gly Val Phe Gly Thr Met Leu Asp Gly Glu Gin 485 490 495 Ala Arg Leu Asn Pro Ala Ser Ala Val Phe Asp Ser Thr Ser Arg Ser 500 505 510 Arg He Thr Arg Val Ser Ser Ser Ser He Lys Ala Ala Tyr Thr Thr 515 520 525 Ser Thr Cys Phe Lys Val Val Lys Thr Asn Lys Thr Tyr Cys Leu Ser 530 535 540 He Ala Glu He Ser Asn Thr Leu Phe Gly Glu Phe Arg He Val Pro 545 550 555 560 Leu Leu Val Glu He Leu Lys Asp Asp Gly Val Arg Glu Ala Arg Ser 565 570 575 Gly (2) INFORMATION FOR SEQ ID NO:31: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 57 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: circular (ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (vi) ORIGINAL SOURCE: (A) ORGANISM: Plasmid (vii) IMMEDIATE SOURCE: (B) CLONE: 538-46.26 (Junction A) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:31: CACATACGAT TTAGGTGACA CTATAGAATA CAAGCTTTAT ACCATTATAG ATACATT (2) INFORMATION FOR SEQ ID NO:32: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 108 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: circular (ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (vi) ORIGINAL SOURCE: (A) ORGANISM: Plasmid (vii) IMMEDIATE SOURCE: (B) CLONE: 538-46.26 (Junction B) (ix) FEATURE: (A) NAME/KEY: exon (B) LOCATION: 88..102 (D) OTHER INFORMATION: /codon_Start= 88 /functions "Translational start of hybrid protein" /products "N-terminal peptide" /numbers l /standard_name= "Translation of synthetic DNA sequence" (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 103..108 (C) IDENTIFICATION METHOD: experimental (D) OTHER INFORMATION: /partial /codon_start= 103 /products "NDV Heamagglutinin-Neuraminidase" /evidences EXPERIMENTAL /gene= "HN" /numbers 2 (xi) SEQUENCE DESCRIPTION: SEC ID NO:32: ET CATGTAGTCG ACTCTAGAAA AAATTGAAAA ACTATTCTAA TTTATTGCAC GGAGATCTTT 60 ττττττττττ TTTTTTTTGG CATATAAATG AATTCGGATC CG GAC CGC 108 Asp Arg 1 (2) INFORMATION FOR SEQ ID NO:33: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 2 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:33: Asp Arg 1 (2) INFORMATION FOR SEQ ID NO:34: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 108 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: circular (ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (vi) ORIGINAL SOURCE: (A) ORGANISM: Plasmid (vii) IMMEDIATE SOURCE: (B) CLONE: 538-46.26 (Junction C) (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 70..84 (D) OTHER INFORMATION: /codon_start= 70 /functions "Translational start of hybrid protein" /products "N-terminal peptide" /numbers i /s andard_names "Translation of synthetic DNA sequence" (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 85..108 (C) IDENTIFICATION METHOD: experimental (D) OTHER INFORMATION: /partial /codon_start= 85 /functions "marker enzyme" /products "Beta-galactosidase" /evidences EXPERIMENTAL /genes "lacZ" /numbers 2 /citations ([1]) (x) PUBLICATION INFORMATION: (A) AUTHORS: Ferrari, Franco A Trach, Kathleen Hoch, James A (B) TITLE: Sequence Analysis of the spoOB Locus Reveals a Polycistronic Transcription Unit (C) JOURNAL: J. Bacteriol. (D) VOLUME: 161 (E) ISSUE: 2 (F) PAGES: 556-562 (G) DATE: Feb.-1985 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:34: TGCGACATCA AGAATCAAAC CGAATGCCCT CGACTCTAGA ATTTCATTTT GTTTTTTTCT ATGCTATAA ATG AAT TCG GAT CCC GTC GTT TTA CAA CGT CGT GAC TGG 1 Met Asn Ser Asp Pro Val Val Leu Gin Arg Arg Asp Trp 1 5 1 5 (2) INFORMATION FOR SEQ ID NO:35: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 5 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:35: Met Asn Ser Asp Pro 1 5 (2) INFORMATION FOR SEQ ID NO:36: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 8 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:36: Val Val Leu Gin Arg Arg Asp Trp 1 5 (2) INFORMATION FOR SEQ ID NO:37: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 108 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: circular (ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (vi) ORIGINAL SOURCE: .. . _, ,__.-. (A) ORGANISM: Plasmid (vii) IMMEDIATE SOURCE: (B) CLONE: 538-46.26 (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 1..54 (C) IDENTIFICATION METHOD: experimental (D) OTHER INFORMATION: /partial /codon_start = l /functions "marker enzyme" /products "Beta-galactosidase" /evidences EXPERIMENTAL /gene= "lacZ" /numbers l /citation s ([1]) (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: 55..63 (C) IDENTIFICATION METHOD: experimental (D) OTHER INFORMATION: /codon_start= 55 /functions "Translational finish of hybrid protein" /products "C-terminal peptide" /evidences EXPERIMENTAL /number= 2 /standard_name= "Translation of synthetic DNA sequence" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:37: GAA ATC CAG CTG AGC GCC GGT CGC TAC CAT TAC CAG TTG GTC TGG TGT 48 Glu He Gin Leu Ser Ala Gly Arg Tyr His Tyr Gin Leu Val Trp Cys 1 5 10 15 CAA AAA GAT CCA TAATTAATTA ACCCGGGTCG AGGGTCGAAG ACCAAATTCT 100 Gin Lys Asp Pro 1 AACATGGT 108 (2) INFORMATION FOR SEQ ID NO:38: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 18 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:38: Glu He Gin Leu Ser Ala Gly Arg Tyr His Tyr Gin Leu Val Trp Cys 1 5 10 15 Gin Lys (2) INFORMATION FOR SEQ ID NO:39: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 2 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:39: Asp Pro 1 (2) INFORMATION FOR SEQ ID NO:40: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 57 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: double (D) TOPOLOGY: circular (ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (vi) ORIGINAL SOURCE: (A) ORGANISM: Plasmid (vii) IMMEDIATE SOURCE: (B) CLONE: 538-46.26 (Junction E) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:40: AGATCCCCGG GCGAGCTCGA ATTCGTAATC ATGGTCATAG CTGTTTCCTG TGTGAAA (2) INFORMATION FOR SEQ ID NO:41: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 27 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: N (iv) ANTI-SENSE: N (vi) ORIGINAL SOURCE: (A) ORGANISM: Pseudorabies virus \ Synthetic oligonucleotide p (xi) SEQUENCE DESCRIPTION: SEQ ID NO:41: CGCGAATTCG CTCGCAGCGC TATTGGC (2) INFORMATION FOR SEQ ID NO:42: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: N (iv) ANTI-SENSE: N (vi) ORIGINAL SOURCE: (A) ORGANISM: Pseudorabies virus \ Synthetic oligonucleotide pr (xi) SEQUENCE DESCRIPTION: SEQ ID NO:42: GTAGGAGTGG CTGCTGAAG 1

Claims: hat is claimed is: 1. A recombinant swinepox virus capable of replication in an animal into which the recombinant swinepox virus is introduced which comprises swinepox viral DNA and foreign DNA, encoding RNA which does not naturally occur in the animal into which the recombinant swinepox virus is introduced, the foreign DNA being inserted into the swinepox viral DNA at an insertion site which is not essential for replication of the swinepox virus and being under the control of a promoter. 2. A recombinant swinepox virus of claim 1, wherein the RNA encodes a polypeptide. 3. A recombinant swinepox virus of claim 2, wherein the polypeptide is antigenic in the animal. 4. A recombinant swinepox virus of claim 1, wherein the insertion site is present within the larger Hindlll to Bglll subfragment of the Hindlll M fragment of the swinepox virus. 5. A recombinant swinepox virus of claim 4, wherein the insertion site is within an open reading frame contained in the Hindlll to Bglll subfragment. 6. A recombinant swinepox virus of claim 5, wherein the insertion site is the AccI restriction endonuclease site located in the Hindlll to Bglll subfragment. 7. A recombinant swinepox virus of claim 1, wherein the insertion site is within an open reading frame encoding swinepox thymidine kinase. 8. A recombinant swinepox virus of claim 3, wherein the antigenic polypeptide is or is from PRV glycoprotein 50, PRV glycoprotein II, PRV glycoprotein III, PRV 5 glycoprotein H, TGE glycoprotein 195, TGE matrix protein, swine rotavirus glycoprotein 38, swine parvovirus capsid protein, Serpulina hyodysenteriae protective antigen, BVD glycoprotein 55, NDV hemagglutinin-neura inidase, swine flu hemagglutinin 10 and swine flu neuraminidase. 9. A recombinant swinepox virus of claim 3, wherein the antigenic polypeptide is or is from Serpulina hyodysenteriae , Foot and Mouth Disease Virus, Hog 15 Cholera Virus, Swine Influenza Virus, African Swine Fever Virus or Mycoplasma hyopneumoniae. 10. A recombinant swinepox virus of claim 8, wherein the antigenic polypeptide is PRV glycoprotein 50. 20 11. A recombinant swinepox virus of claim 8, wherein the antigenic polypeptide is NDV hemagglutinin- neuraminidase. 25 12. A recombinant swinepox virus of claim 2, wherein the polypeptide is a detectable marker. 13. A recombinant swinepox virus of claim 2 , wherein the polypeptide is E . coli β-galactosidase. 30 14. A recombinant swinepox virus of claim 13, designated S-SPV-003 (ATCC Accession No. VR 2335) . 15. A recombinant swinepox virus of claim 10 further 5 comprising foreign DNA encoding a polypeptide which is a detectable marker. HEET 16. A recombinant swinepox virus of claim 15 designated S- SPV-008 (ATCC Accession No. VR 2339) . 17. A recombinant swinepox virus of claim 11 further comprising foreign DNA encoding a polypeptide which is a detectable marker. 18. A recombinant swinepox virus of claim 17, designated S-SPV-009 (ATCC Accession No. VR 2344) . 19. A recombinant swinepox virus of claim 1, wherein the promoter is a swinepox viral promoter. 20. A recombinant swinepox virus of claim 1, wherein the promoter is a synthetic pox viral promoter. 21. A homology vector for producing a recombinant swinepox virus by inserting foreign DNA into the genomic DNA of a swinepox virus which comprises a double-stranded DNA molecule consisting essentially of: a) double-stranded foreign DNA encoding RNA which does not naturally occur in an animal into which the recombinant swinepox virus is introduced; b) at one end of the foreign DNA, double-stranded swinepox viral DNA homologous to genomic DNA located at one side of a site on the genomic DNA which is not essential for replication of the swinepox virus; and c) at the other end of the foreign DNA, double- stranded swinepox viral DNA homologous to genomic DNA located at the other side of the same site on the genomic DNA. SUBSTITUTE H 22. A homology vector of claim 21, wherein the RNA encodes a polypeptide. 23. A homology vector of claim 22, wherein the polypeptide 5 is antigenic in the animal. 24. A homology vector of claim 23, wherein the antigenic polypeptide is or is from PRV glycoprotein 50, PRV glycoprotein II, PRV glycoprotein III, PRV 10 glycoprotein H, TGE glycoprotein 195, TGE matrix protein, swine rotavirus glycoprotein 38, swine parvovirus capsid protein, Serpulina hyodysenteriae protective antigen, BVD glycoprotein 55, NDV hemagglutinin-neuraminidase, swine flu hemagglutinin 15 or swine flu neuraminidase. 25. A homology vector of claim 23, wherein the antigenic polypeptide is or is from Serpulina hyodysenteriae , Foot and Mouth Disease Virus, Hog Cholera Virus, Swine 20 Influenza Virus, African Swine Fever Virus or Mycoplasma hyopneumoniae . 26. A homology vector of claim 22, wherein the polypeptide is a detectable marker. 25 27. A homology vector of claim 22, wherein the polypeptide is E. coli β-galactosidase. 28. A homology vector of claim 21, wherein the double- 30 stranded swinepox viral DNA is homologous to genomic DNA present within the larger Hindlll to Bglll subfragment of the Hindlll M fragment of swinepox virus. 35 29. A homology vector of claim 28, wherein the double- stranded swinepox viral DNA is homologous to genomic DNA present within the open reading frame contained in the Hindlll to Bglll subfragment. 30. A homology vector of claim 29, wherein the double- stranded swinepox viral DNA is homologous to genomic DNA present within the AccI restriction endonuclease site located in the Hindlll to Bglll subfragment. 31. A homology vector of claim 21, wherein the double- stranded swinepox viral DNA is homologous to genomic DNA present within the open reading frame encoding swinepox thymidine kinase. 32. A homology vector of claim 21, wherein the foreign DNA further comprises a synthetic pox viral promoter. 33. A vaccine which comprises an effective immunizing amount of the recombinant swinepox virus of claims 3, 8, 9, 16 or 18 and a suitable carrier. 34. A vaccine which comprises an effective immunizing amount of the recombinant swinepox virus of claim 16 and a suitable carrier. 35. A method of immunizing an animal which comprises administering to the animal an effective immunizing dose of the vaccine of claim 33. 36. A method of immunizing an animal which comprises administering to the animal an effective immunizing dose of the vaccine of claim 34. 37. A method of claim 35 or 36, wherein the animal is a swine, bovine equine, caprine, or ovine. 38. A method for testing a swine to determine whether the swine has been vaccinated with the vaccine of claim 33 or is infected with a naturally-occurring pseudorabies virus which comprises: a) obtaining from the swine to be tested a sample of a suitable body fluid; b) detecting in the sample the presence of antibodies to pseudorabies virus, the absence of such antibodies indicating that the swine has been neither vaccinated nor infected; and c) for the swine in which antibodies to pseudorabies virus are present, detecting in the sample the absence of antibodies to pseudorabies virus antigens which are normally present in the body fluid of a swine infected by the naturally-occurring pseudorabies virus but which are not present in a vaccinated swine, the absence of such anti-bodies indicating that the swine was vaccinated and is not infected. 39. A host cell infected with the recombinant swinepox virus of claim l. 40. A host cell of claim 39, wherein the host cell is a mammalian cell. 41. A host cell of claim 40, wherein the mammalian cell is a Vero cell. 42. A host cell of claim 41, wherein the mammalian cell is an EMSK cell.

Inventors: COCHRAN MARK D, JUNKER DAVID E, COCHRAN, MARK, D., JUNKER, DAVID, E.
Applicants: SYNTRO CORP, SYNTRO CORPORATION
Application year: 1993
Main classification: C12N7
IPC codes: A61K39, A61P31, C07K14, C12N5, C12N7, C12N15, C12P21, C12Q1, C12R1