AU4863396A - Recombinant swinepox virus - Google Patents

Description

RECOMBINANT SWINEPOX VIRUS

This application is a continuation-in-part of U.S. Serial No. 08/480,640, filed June 7, 1995, U.S. Serial No. 08/488,237, filed June 7, 1995, U.S. Serial No. 08/472,679, filed June 7, 1995, and- a continuation-in- part of U.S. Serial No. 08/375,992, filed January 19, 1995, the contents of which are incorporated by reference into the present application.

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. The disclosures of these publications are hereby incorporated by reference into this application.

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 the cytoplasm of the host cell. SPV is the only member of the genus Suipoxvirus . 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 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, 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. SPV is only mildly pathogenic, being characterized by a self-limiting infection with lesions detected only in the skin 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 fro the genus Suipoxvirus . 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

This invention provides a recombinant swinepox virus comprising a foreign DNA sequence inserted into the swinepox virus genomic DNA, wherein the foreign DNA sequence is inserted within a HindiII N fragment of the swinepox virus genomic DNA and is capable of being expressed in a swinepox virus infected host cell.

The invention further provides homology vectors, vaccines and methods of immunization.

BRIEF DESCRIPTION OF THE INVENTION

Figures 1A - IB:

Show a detailed diagram of SPV genomic DNA (Kasza strain) including the unique long and Terminal repeat (TR) regions. A restriction map for the enzyme Hindlll 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.

Figures 2A - 2B:

Show the 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 NO:l) covers a 599 base pair sequence which flanks the unique AccI site as indicated in Figures 3A-3C. 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 HindiII site as indicated in Figures 3A-3C. The beginning (Asp) and end (lie) of a 220 amino acid ORF is indicated by the translation of amino acids below the ENA sequence.

Figures 3A - 3C:

Show the homology which exists between the 515.85.1 ORF and the Vaccinia virus OIL ORF. Figure 3A shows two maps: The first line of Figure 3A is a restriction map of the SPV Hindi11 M fragment and the second is a restriction map of the DΝA insertion in plasmid 515-85.1. The location of the 515-85.1 [W OIL-like] ORF is also indicated on the map. The locations of the DΝA sequences shown in Figures 3B and 3C are indicated below the map by heavy bars in Figure 3A. Figure 3B shows the homology between the W OIL ORF (SEQ ID NO:5) and the 515-85.1 ORF (SEQ ID NO:6) at their respective N-termini. Figure 3C 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.

Figures 4A - 4D:

Show a description of the DNA insertion in Homology Vector 520-17.5. Figure 4A contains a diagram showing the orientation of DNA fragments assembled in plasmid 520-17.5 and table indicating the origin of each fragment. Figure 4B shows the sequences located at each of the junctions A and B between fragments, and Figure 4C shows the sequences located at Junctions C and D (SEQ ID NO'S: 9, 10, 13, and 16) . Figures 4B and 4C further describe 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 0 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 1 (EP1) , late promoter 2 (LP2) , lactose operon Z gene (lacZ) , and

Escherichia coli (E. coli) .

Figures 5A - 5D:

Show a detailed description of the DNA insertion in Homology Vector 538-46.16. Figure 5A contains a diagram showing the orientation of DNA fragments assembled in plasmid 538-46.16 and a table indicating the origin of each fragment . Figure 5B shows the sequences located at Junctions A and B between fragments, Figure 5C shows sequences located at Junction C and Figure 5D shows sequences located at Junctions D and E (SEQ ID NO'S: 17, 18, 21, 26, and 28) . Figures 5B to 5D also describe 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) , g50 (gD) , glycoprotein 63 (g63) , early promoter 1 (EP1) , late promoter 1 (LP1) (SEQ ID NO: 46) , 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 S62/26) 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-Neuraminidase 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.

Figures 8A - 8D; Show a detailed description of the DNA insertion in Homology Vector 538-46.26. Figure 5A contains a diagram showing the orientation of DNA fragments assembled in plasmid 538-46.26 and table indicating the origin of each fragment . Figure 8B shows the sequences located at Junctions A and B between fragments; Figure 8C shows the sequences located at Junctions C and D, Figure 8D shows the sequences located at Junction E (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 in Figures 8B and 8D. 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 ) . Figures 9A - 9C ;

Show a detailed description of Swinepox Virus S-SPV-

010 and the DNA insertion in Homology Vector 561- 36.26. Figure 9A contains a diagram showing the orientation of DNA fragments assembled in plasmid 561-36.26 and a table indicating the origin of each fragment . Figure 9B shows the sequences located at Junctions A and B between fragments and Figure 9C show the sequences located at junction C and D (SEQ ID. NO: 47, 48, 49,50) . The restriction sites used to generate each fragment as well as synthetic linker sequences which are used to join the fragments are described for each junction in Figures 9B and 9C. The location of several gene coding regions and regulatory elements is also given. The following two conventions are used: numbers in parentheses, (), refer to amino acids, and restriction sites in brackets, [] , indicate the remnants of sites which are destroyed during construction. The following abbreviations are used: swinepox virus (SPV) , Escherichia coli (E. col i ) , thymidine kinase (TK) , pox synthetic late promoter 1 (LP1) , base pairs (BP) .

Figures 10A - 10D:

Show a detailed description of Swinepox Virus S-SPV-

011 and the DNA insertion in Homology Vector 570- 91.21. Figure 10A contains a diagram showing the orientation of DNA fragments assembled in plasmid 570-91.21 and a table indicating the origin of each fragment. Figure 10B show the sequences located at Junctions A and B between fragments; Figure 10C shows the sequences located at Junction C, and Figure 10D shows the sequences located at Junctions 10D and 10E(SEQ ID NOs : 51, 52, 53, 54, 55) . The restriction sites used to generate each fragment as well as synthetic linker sequences which are used to join the fragments are described for each junction in Figures 10B to 10D. The location of several gene coding regions and regulatory elements is also given. The following two conventions are used: numbers in parentheses, (), refer to amino acids, and restriction sites in brackets, [] , indicate the remnants of sites which are destroyed during construction. The following abbreviations are used: swinepox virus (SPV) , pseudorabies virus (PRV) , Escherichia coli (E. coli) , pox synthetic late promoter 1 (LP1) , pox synthetic early promoter 2

(EP2) (SEQ ID NO: 45), gill (gC) , base pairs (BP) .

Figures 11A - IIP: Show a detailed description of Swinepox Virus S-SPV- 012 and the DNA insertion in Homology Vector 570- 91.41. Figure 11A contains a diagram showing the orientation of DNA fragments assembled in plasmid 570-91.41 and a table indicating the origin of each fragment. Figure 11B shows the sequences located at Junctions A and B between fragments, Figure 11C shows the sequences located at Junction C, and Figure 11D shows the sequence located at Junctions D and E. (SEQ ID NOs: 56, 57, 58, 59, 60). The restriction sites used to generate each fragment as well as synthetic linker sequences which are used to join the fragments are described for each junction in Figures 11B to 11D. The location of several gene coding regions and regulatory elements is also given. The following two conventions are used: numbers in parentheses, () , refer to amino acids, and restriction sites brackets, [] , indicate the remnants of sites which are destroyed during construction. The following abbreviations are used: swinepox virus (SPV) , pseudorabies virus (PRV) , Escherichia coli (E. coli ) , pox synthetic late promoter 1 (LP1) , pox synthetic early promoter 1 late promoter 2 (EP1LP2) (SEQ ID NO: 43) , gill (gC) , base pairs (BP) .

Figures 12A - 12D: Show a detailed description of Swinepox Virus S-PRV-

013 and the DNA insertion in Homology Vector 570- 91.64. Figure 12A contains a diagram showing the orientation of DNA fragments assembled in plasmid 570-91.64 and a table indicating the origin of each fragment. Figure 12B shows the sequences located at Junctions A and B between fragments, Figure 12C shows the sequences located at Junction C, and Figure 12D shows the sequences located at Junctions D and E (SEQ ID NOs: 61, 62, 63, 64, 65) . The restriction sites used to generate each fragment as well as synthetic linker sequences which are used to join the fragments are described for each junction in Figures 12B to 12D. The location of several gene coding regions and regulatory elements is also given. The following two conventions are used: numbers in parentheses, () , refer to amino acids, and restriction sites in brackets, [] , indicate the remnants of sites which are destroyed during construction. The following abbreviations are used: swinepox virus (SPV) , pseudorabies virus (PRV) , Escherichia coli (E. coli ) , pox synthetic late promoter 1 (LP1) , pox synthetic late promoter 2 early promoter 2 (LP2EP2) (SEQ ID NO: 44) , gill (gC) base pairs (BP) .

Figures 13A - 13D:

Show a detailed description of Swinepox Virus S-PRV-

014 and the DNA insertion in Homology Vector 599- 65.25. Figure 13A contains a diagram showing the orientation of DNA fragments assembled in plasmid 599-65.25 and a table indicating the origin of each fragment. Figure 13B shows sequences located at Junctions A and B between the fragments, Figure 13C shows sequences located at Junction C, and Figure 13D shows sequences located at Junctions D and E. (SEQ ID NOs: 66, 67, 68, 69, and 70) . The restriction sites used to generate each fragment as well as synthetic linker sequences which are used to join the fragments are described for each junction in Figures 13B to 13D. The location of several gene coding regions and regulatory elements is also given. The following two conventions are used: numbers in parentheses, 0 , refer to amino acids, and restriction sites in brackets, [] , indicate the remnants of sites which are destroyed during construction. The following abbreviations are used: swinepox virus (SPV) , infectious laryngotracheitis virus (ILT) , Escherichia coli (E. coli ) , pox synthetic late promoter 1 (LP1) , pox synthetic early promoter 1 late promoter 2 (EP1LP2) , glycoprotein G

(gG) , polymerase chain reaction (PCR) , base pairs (BP) .

Figures 14A - 14D:

Show a detailed description of Swinepox Virus S-SPV- 016 and the DNA insertion in Homology Vector 624- 20. IC. Figure 14A contains a diagram showing the orientation of DNA fragments assembled in plasmid 624-20.1C and a table indicating the origin of each fragment . Figure 14B shows the sequences located at Junctions A and B between fragments; Figure 14C shows the sequences located at Junction C, and Figure 14D shows the sequences at Junctions D and E. (SEQ ID NOs: 71, 72, 73, 74, and 75) . The restriction sites are used to generate each fragment as well as synthetic linker sequences which are used to join the fragments are described for each junction in Figures 14B to 14D. The location of several gene coding regions and regulatory elements is also given. The following two conventions are used: numbers in parentheses, () , refer to amino acids, and restriction sites in brackets, [] , indicate the remnants of sites which are destroyed during construction. The following abbreviations are used: swinepox virus (SPV) , infectious laryngotracheitis virus (ILT) , Escherichia col i (E. coli ) , pox synthetic late promoter 1 (LP1) , pox synthetic late promoter 2 early promoter 2 (LP2EP2) , glycoprotein I (gl) , polymerase chain reaction

(PCR) , base pairs (BP) .

Figures 15A - 15D:

Show a detailed description of Swinepox Virus S-SPV- 017 and the DNA insertion in Homology Vector 614- 83.18. Figure 15A contains a diagram showing the orientation of DNA fragments assembled in plasmid 614-83.18 and a table showing the origin of each fragment . Figure 15B shows the sequences located at Junctions A and B between fragments, Figure 15C shows the sequences at Junction C, and Figure 15D shows the sequences located at Junctions D and E. The restriction sites used to generate each fragment as well as synthetic linker sequences which are used to join the fragments are described for each junction in Figures 15B to 15D. The location of several gene coding regions and regulatory elements is also given. The following two conventions are used: numbers in parentheses, () , refer to amino acids, and restriction sites in brackets, [] , indicate the remnants of sites which are destroyed during construction. The following abbreviations are used: swinepox virus (SPV) , infectious bovine rhinotracheitis virus (IBR) , Escherichia coli (E. col i ) , pox synthetic late promoter 1 (LP1) , pox synthetic late promoter 2 early promoter 2 (LP2EP2) , glycoprotein G (gG) , polymerase chain reaction (PCR) , base pairs (BP) .

Figure 16: Western blot of lysates from recombinant SPV infected cells with polyclonal goat anti-PRV gill (gC) . Lanes (A) S-PRV-002 (U.S. Patent No. 4,877,737, issued October 31, 1989) infected cell lysate, (B) molecular weight markers, (C) mock- infected EMSK cell lysate, (D) S-SPV-003 infected cell lysate, (E) S-SPV-008 infected cell lysate, (F) S-SPV-011 infected cell lysate, (G) S-SPV-012 infected cell lysate, (H) S-SPV-013 infected cell lysate. Cell lysates are prepared as described in the PREPARATION OF INFECTED CELL LYSATES. Approximately 1/5 of the total lysates sample is loaded in each lane.

Figure 17: Map showing the 5.6 kilobase pair Hindi11 M swinepox virus genomic DNA fragment . Open reading frames

(ORF) are shown with the number of amino acids coding in each open reading frame. The swinepox virus ORFs show significant sequence identities to the vaccinia virus ORFs and are labeled with the vaccinia virus nomenclature (56 and 58) . The I4L

ORF (SEQ ID NO: 196) shows amino acid sequence homology to ribonucleotide reductase large subunit

(57) , and the OIL ORF (SEQ ID NO: 193) shows amino acid sequence homology to a leucine zipper motif characteristic of certain eukaryotic transcriptional regulatory proteins (13) . The Bglll site in the I4L ORF and the AccI site in the OIL ORF are insertion sites for foreign DNA into non-essential regions of the swinepox genome. The homology vector 738-94.4 contains a deletion of SPV DNA from nucleotides 1679 to 2452 (SEQ ID NO: 189) . The black bar at the bottom indicates regions for which the DNA sequence is known and references the SEQ ID NOs : 189 and 195. Positions of restriction sites AccI, Bglll, and Hindlll are shown. I3L ORF (SEQ ID NO: 190) , I2L ORF (SEQ ID NO: 191) and E10R ORF (SEQ ID NO: 194) are shown. SEQ ID NO 221 contains the complete 5785 base pair sequence of the Hindi11 M fragment. Open reading frames within the SPV HindiII M fragment are the partial I4L ORF (445 AA; Nucl 2 to 1336) ; I3L ORF (275 AA; Nucl 1387 to 2211) ; I2L ORF (75 AA; Nucl 2215 to 2439) ; I1L ORF (313 AA; Nucl 2443 to 3381) ; OIL ORF (677 AA; Ncl 3520 to 5550) ; partial E10R ORF (64 AA; Nucl 5787 to 5596) .

Figures 18A - 18D:

Show a detailed description of Swinepox Virus S-SPV- 034 and the DNA insertion in Homology Vector 723- 59A9.22. Figure 18A contains a diagram showing the orientation of DNA fragments assembled in plasmid 723-59A9.22 and a table indicating the origin of each fragment. Figure 18B shows the sequences located at Junctions A and B between fragments, Figure 18C shows the sequences located at Junction C, and Figure 18D shows the sequences located at Junctions D and E. The restriction sites used to generate each fragment as well as synthetic linker sequences which are used to join the fragments are described for each junction in Figures 18B to 18D. The location of several gene coding regions and regulatory elements is also given. The following two conventions are used: numbers in parentheses, () , refer to amino acids, and restriction sites in brackets, [] , indicate the remnants of sites which are destroyed during construction. The following abbreviations are used: swinepox virus (SPV) , equine influenza virus (EIV) , Escherichia coli (E. coli ) , pox synthetic late promoter 1 (LP1) , pox synthetic late promoter 2 early promoter 2 (LP2EP2) , neuraminidase (NA) , Prague (PR) , polymerase chain reaction (PCR) , base pairs (BP) .

Figures 19A - 19D;

Show a detailed description of Swinepox Virus S-SPV- 015 and the DNA insertion in Homology Vector 727- 54.60. Figure 19A contains a diagram showing the orientation of DNA fragments assembled in plasmid 727-54.60 and a table indicating the origin of each fragment. Figure 19B shows the sequences located at Junctions A and B between fragments, Figure 19C shows the sequences located at Junction C, and Figure 19D shows the sequences located at Junctions D and E. The restriction sites used to generate each fragment as well as synthetic linker sequences which are used to join the fragments are described for each junction in Figures 19B to 19D. The location of several gene coding regions and regulatory elements is also given. The following two conventions are used: numbers in parentheses, () , refer to amino acids, and restriction sites in brackets, [] , indicate the remnants of sites which are destroyed during construction. The following abbreviations are used: swinepox virus (SPV) , pseudorabies virus (PRV) , Escherichia coli (E. coli ) , pox synthetic late promoter 1 (LP1) , pox synthetic late promoter 2 early promoter 2 (LP2EP2) , glycoprotein B (gB) , base pairs (BP) .

Figures 20A - 20D:

Show a detailed description of Swinepox Virus S-SPV- 031 and the DNA insertion in Homology Vector 727- 67.18. Figure 20A contains a diagram showing the orientation of DNA fragments assembled in plasmid 727-67.18 and a table indicating the origin of each fragment. Figure 2OB shows the sequences located at Junctions A and B between fragments, Figure 20C shows the sequences located at Junction C, and Figure 20D shows the sequences located at Junctions D and E. The restriction sites used to generate each fragment as well as synthetic linker sequences which are used to join the fragments are described for each junction in Figures 20B to 20D. The location of several gene coding regions and regulatory elements is also given. The following two conventions are used: numbers in parentheses, () , refer to amino acids, and restriction sites in brackets, [] , indicate the remnants of sites which are destroyed during construction. The following abbreviations are used: swinepox virus (SPV) , Escheri chia coli (E. coli ) , pox synthetic late promoter 1 (LP1) , pox synthetic early promoter 1 late promoter 2 (EP1LP2) , antigen (Ag) , base pairs (BP) .

Figures 21A - 2ID;

Show a detailed description of Swinepox Virus S-SPV- 033 and the DNA insertion in Homology Vector 732- 18.4. Figure 21A contains a diagram showing the orientation of DNA fragments assembled in plasmid 732-18.4 and a table indicating the origin of each fragment. Figure 21B shows the sequences located at Junctions A and B between fragments, Figure 21C shows the sequences located at Junction C, and Figure 21D shows the sequences located at Junctions D and E. The restriction sites used to generate each fragment as well as synthetic linker sequences which are used to join the fragments are described for each junction in Figures 21B to 21D. The location of several gene coding regions and regulatory elements is also given. The following two conventions are used: numbers in parentheses, () , refer to amino acids, and restriction sites in brackets, [] , indicate the remnants of sites which are destroyed during construction. The following abbreviations are used: swinepox virus (SPV) , Escherichia coli (E. coli) , pox synthetic late promoter 1 (LPl) , pox synthetic late promoter 2 early promoter 2 (LP2EP2) , equine influenza virus (EIV) , neuraminidase (NA) , Alaska (AK) , polymerase chain reaction (PCR) , base pairs (BP) .

Figures 22A - 22C:

Show a detailed description of Swinepox Virus S-SPV- 036 and the DNA insertion in Homology Vector 741- 80.3. Figure 22A contains a diagram showing the orientation of DNA fragments assembled in plasmid 741-80.3 and a table indicating the origin of each fragment. Figure 22B shows the sequences located at Junctions A, B, and C between fragments and Figure 22C shows the sequences located at Junctions D, E and F. The restriction sites used to generate each fragment as well as synthetic linker sequences which are used to join the fragments are described for each junction in Figures 22B and 22C. The location of several gene coding regions and regulatory elements is also given. The following two conventions are used: numbers in parentheses, () , refer to amino acids, and restriction sites in brackets, [] , indicate the remnants of sites which are destroyed during construction. The following abbreviations are used: swinepox virus (SPV) , pseudorabies virus (PRV) , Escherichia coli (E. coli) , human cytomegalovirus immediate early (HCMV IE) , pox synthetic late promoter 1 (LPl) , pox synthetic late promoter 2 early promoter 2 (LP2EP2) , polyadenylation site (poly A) , base pairs (BP) .

Figures 23A - 23D; Show a detailed description of Swinepox Virus S-SPV- 035 and the DNA insertion in Homology Vector 741- 84.14. Figure 23A contains a diagram showing the orientation of DNA fragments assembled in plasmid 741-84.14 and a table indicating the origin of each fragment. Figure 23B shows the sequences located at Junctions A and B between fragments, Figure 23C shows the sequences located at Junction C, and Figure 23D shows the sequences located at Junctions D and E. The restriction sites used to generate each fragment as well as synthetic linker sequences which are used to join the fragments are described for each junction in Figures 23B to 23D. The location of several gene coding regions and regulatory elements is also given. The following two conventions are used: numbers in parentheses, () , refer to amino acids, and restriction sites in brackets, [] , indicate the remnants of sites which are destroyed during construction. The following abbreviations are used: swinepox virus (SPV) , pseudorabies virus (PRV) , Escherichia coli (E . coli ) , pox synthetic late promoter 1 (LPl) , pox synthetic late promoter 2 early promoter 2 (LP2EP2) , interleukin-2 (IL-2) , glycoprotein X (gX) polymerase chain reaction (PCR) , sequence (seq) , base pairs (BP) .

Figures 24A - 24D;

Show a detailed description of Swinepox Virus S-SPV- 038 and the DNA insertion in Homology Vector 744-34. Figure 24A contains a diagram showing the orientation of DNA fragments assembled in plasmid 744-34 and a table indicating the origin of each fragment. Figure 24B shows the sequences located at Junction A and B between fragments, Figure 24C shows the sequences located at Junction C, and Figure 24D shows the sequences located at Junctions D and E. The restriction sites used to generate each fragment as well as synthetic linker sequences which are used to join the fragments are described for each junction in Figures 24B and 24D. The location of several gene coding regions and regulatory elements is also given. The following two conventions are used: numbers in parentheses, () , refer to amino acids, and restriction sites in brackets, [] , indicate the remnants of sites which are destroyed during construction. The following abbreviations are used: swinepox virus (SPV) , equine herpesvirus type 1 (EHV-1) , Escherichia coli (E. coli ) , pox synthetic late promoter 1 (LPl) , pox synthetic late promoter 2 early promoter 2 (LP2EP2) , glycoprotein B (gB) , polymerase chain reaction (PCR) , base pairs (BP) .

Figures 25A - 25D;

Show a detailed description of Swinepox Virus S-SPV- 039 and the DNA insertion in Homology Vector 744-38.

Figure 25A contains a diagram showing the orientation of DNA fragments assembled in plasmid 744-38 and a table indicating the origin of each fragment. Figure 25B shows the sequences located at Junction A and B between fragments. Figure 25C shows the sequences located at Junction C and Figure 25D shows the sequences located at Junctions D and E. The restriction sites used to generate each fragment as well as synthetic linker sequences which are used to join the fragments are described for each junction in Figures 25B to 25D. The location of several gene coding regions and regulatory elements is also given. The following two conventions are used: numbers in parentheses, () , refer to amino acids, and restriction sites in brackets, [] , indicate the remnants of sites which are destroyed during construction. The following abbreviations are used: swinepox virus (SPV) , equine herpesvirus type 1 (EHV-l) , Escherichia coli

(E. coli ) , pox synthetic late promoter 1 (LPl) , pox synthetic late promoter 2 early promoter 2 (LP2EP2) , glycoprotein D (gD) , polymerase chain reaction

(PCR) , base pairs (BP) .

Figures 26A-26D;

Detailed description of Swinepox Virus S-SPV-042 and the DNA insertion in Homology Vector 751-07.A1. Diagram showing the orientation of DNA fragments assembled in plasmid 751-07.Al. The origin of each fragment is indicated in the table. The sequence located at each of the junctions between fragments is also shown. The restriction sites used to generate each fragment as well as synthetic linker sequences which are used to join the fragments are described for each junction. Figures 26A-26D show the sequences located at Junction A (SEQ ID NOS: 197) , (SEQ ID NO: 198) , C (SEQ ID NO: 199), D (SEQ ID NO: 200) and E (SEQ ID NO: 201) between fragments and the sequences located at the junctions. The location of several gene coding regions and regulatory elements is also given. The following two conventions are used: numbers in parentheses,

() , refer to amino acids, and restriction sites in brackets, [] , indicate the remnants of sites which are destroyed during construction. The following abbreviations are used: swinepox virus (SPV) , chicken interferon (cIFN) , Escherichia coli (E. coli) , pox synthetic late promoter 1 (LPl) , pox synthetic late promoter 2 early promoter 2 (LP2EP2) , polymerase chain reaction (PCR) , base pairs (BP) .

Figures 27A-27D;

Detailed description of Swinepox Virus S-SPV-043 and the DNA insertion in Homology Vector 751-56.Al. Diagram showing the orientation of DNA fragments assembled in plasmid 751-56.Al. The origin of each fragment is indicated in the table. The sequences located at each of the junctions between fragments is also shown. Figures 27A-27D show the sequences located at Junction A (SEQ ID NOS: 202) , (SEQ ID NO: 203) , C (SEQ ID NO: 204) , D (SEQ ID NO: 205) and E (SEQ ID NO: 206) between fragments and the sequences located at the junctions. The restriction sites used to generate each fragment as well as synthetic linker sequences which are used to join the several gene coding regions and regulatory elements is also given. The following two conventions are used: numbers in parentheses, (), refer to amino acids, and restriction sites in brackets, [] , indicate the remnants of sites which are destroyed during construction. The following abbreviations are used: swinepox virus (SPV) , chicken myelomonocytic growth factor (cMGF) , Escherichia coli (E. coli) , pox synthetic late promoter 1 (LPl) , pox synthetic late promoter 2 early promoter 2 (LPE2EP2) , polymerase chain reaction (PCR) , base pairs (BP) .

Figure 28A-28D; Detailed description of Swinepox Virus S-SPV-043 and the DNA insertion in Homology Vector 752-22.1. Diagram showing the orientation of DNA fragments assembled in plasmid 752-22.1. The origin of each fragment is indicated in the table. The sequences located at each of the junctions between fragments is also shown. Figures 28A-28D show the sequences located at Junction A (SEQ ID NOS: 207) , (SEQ ID NO: 208) , C (SEQ ID NO: 209) , and D (SEQ ID NO: 210) between fragments and the sequences located at the junctions. The restriction sites used to generate each fragment as well as synthetic linker sequences which are used to join the fragments are described for each junction. The location of several gene coding regions and regulatory elements is also given. The following two conventions are used: numbers in parentheses, () , refer to amino acids, and restrictions sites in brackets, [] , indicate the remnants of sites which are destroyed during construction. The following abbreviations are used: swinepox virus (SPV), Escherichia coli (E. coli) , pox synthetic late promoter 2 early promoter 2 (LP2EP2) , polymerase chain reaction (PCR) , base pairs (BP) .

Figures 29A-29B;

Figure 29A: Restriction Endonuclease Map and Open Reading Frames in the SPV Hindlll N fragment and part of SPV Hindlll M fragment. Insertions of a foreign gene into a non-essential site of the swinepox virus Hind III N and Hind III M genomic DNA include the EcoR V site (S-SPV-060) , SnaB I site (S- SPV-061) , Bgl II site in Hind III N (S-SPV-062) , and the Bgl II site in Hind III M (S-SPV-047) . Insertions of a foreign gene into the I7L ORF (SEQ ID NO. 230) and I4L ORF (SEQ ID NO. 231) indicates that the sequence of the entire open reading frame is non-essential for replication of the swinepox virus and suitable for insertion of foreign genes. Additional sites for insertion of foreign genes include, but are not limited to the two Hind III sites, Ava I site, and the BamHI site.

Figure 29B: Restriction Endonuclease Map and Open Reading Frames in the SPV Hind III K fragment. Insertion of a foreign gene into a non-essential site of the swinepox virus Hind III K genomic DNA include, but is not limited to the EcoR I site (S- SPV-059) . Three open reading frames are identified within a 3.2 kB region of the SPV Hindlll K fragment. Insertions of a foreign gene into the B18R ORF (SEQ ID NO. 228) indicates that the sequence of the entire open reading frame is non-essential for replication of the swinepox virus and suitable for insertion of foreign genes. Also identified is the B4R ORF (SEQ ID NO. 229) which is a site for insertion of a foreign gene. SPV B18R ORF has homology to the vaccinia virus (W) B18R ORF. SPV B18R ORF has more homology to the 77.2 kd protein of rabbit fibroma virus (RFV) . SPV B4R ORF has homology to the vaccinia virus (W) B4R ORF. SPV B4R ORF has more homology to the T5 protein of rabbit fibroma virus (RFV) . The identified open reading frames are within approximately 3200 base pairs of the SPV Hind III K fragment. The remaining approximately 3500 base pairs of the SPV Hind III K fragment has been sequenced previously (R.F. Massung, et al . Virology 197, 511-528 (1993) ) .

Figures 30A-30C;

Detailed description of Swinepox Virus S-SPV-047 and the DNA insertion in Homology Vector 779-94.31. Diagram showing the orientation of DNA fragments assembled in plasmid 779-94.31. The origin of each fragment is indicated in the table. The sequences located at each of the junctions between fragments is also shown. Figures 30A-30C show the sequences located at Junction A (SEQ ID NOS: ) , (SEQ ID NO: ) , C (SEQ ID NO: ) , D (SEQ ID NO: ), and E (SEQ ID NO: ) between fragments and the sequences located at the junctions. The restriction sites used to generate each fragment as well as synthetic linker sequences which are used to join the fragments are described for each junction. The location of several gene coding regions and regulatory elements is also given. The following two conventions are used: numbers in parentheses, () , refer to amino acids, and restrictions sites in brackets, [] , indicate the remnants of sites which are destroyed during construction. The following abbreviations are used: swinepox virus (SPV) , pseudorabies virus (PRV) , Escherichia coli (E. coli) , pox synthetic late promoter 2 early promoter 2 (LP2EP2) , pox synthetic late promoter 1 (LPl) , base pairs (BP) .

Figures 31A-31D:

Detailed description of Swinepox Virus S-SPV-052 and the DNA insertion in Homology Vector 789-41.7. Diagram showing the orientation of DNA fragments assembled in plasmid 789-41.7. The origin of each fragment is indicated in the table. The sequences located at each of the junctions between fragments is also shown. Figures 31A-31D show the sequences located at Junction A (SEQ ID NOS: ) , (SEQ ID NO: ) , C (SEQ ID NO: ) , D (SEQ ID NO: ) , E (SEQ ID NO: ) , and F (SEQ ID NO: ) between fragments and the sequences located at the junctions. The restriction sites used to generate each fragment as well as synthetic linker sequences which are used to join the fragments are described for each junction. The location of several gene coding regions and regulatory elements is also given. The following two conventions are used: numbers in parentheses, () , refer to amino acids, and restrictions sites in brackets, [] , indicate the remnants of sites which are destroyed during construction. The following abbreviations are used: swinepox virus (SPV) , pseudorabies virus (PRV) , Escherichia coli (E. coli) , pox synthetic late promoter 2 early promoter 2 (LP2EP2) , pox synthetic early promoter 1 late promoter 2 (EP1LP2) , pox synthetic late promoter 1 (LPl) , base pairs (BP) . Figures 32A- 32D ;

Detailed description of Swinepox Virus S-SPV-053 and the DNA insertion in Homology Vector 789-41.27. Diagram showing • the orientation of DNA fragments assembled in plasmid 789-41.27. The origin of each fragment is indicated in the table. The sequences located at each of the junctions between fragments is also shown. Figures 32A-32D show the sequences located at Junction A (SEQ ID NOS: ) , (SEQ ID NO: ) , C (SEQ ID NO: ) , D (SEQ ID NO: ) , E (SEQ ID NO: ) , F (SEQ ID NO: ) , and G (SEQ ID NO: ) between fragments and the sequences located at the junctions. The restriction sites used to generate each fragment as well as synthetic linker sequences which are used to join the fragments are described for each junction. The location of several gene coding regions and regulatory elements is also given. The following two conventions are used: numbers in parentheses, () , refer to amino acids, and restrictions sites in brackets, [] , indicate the remnants of sites which are destroyed during construction. The following abbreviations are used: swinepox virus (SPV) , pseudorabies virus (PRV) , Escherichia coli (E. coli) , pox synthetic late promoter 2 early promoter 2 (LP2EP2) , pox synthetic early promoter 1 late promoter 2 (EP1LP2) , pox synthetic late promoter 1 (LPl) , base pairs (BP) .

Figures 33A-33D;

Detailed description of Swinepox Virus S-SPV-054 and the DNA insertion in Homology Vector 789-41.47. Diagram showing the orientation of DNA fragments assembled in plasmid 789-41.47. The origin of each fragment is indicated in the table. The sequences located at each of the junctions between fragments is also shown. Figures 33A-33D show the sequences located at Junction A (SEQ ID NOS: ) , (SEQ ID NO: ) , C (SEQ ID NO: ) , D (SEQ ID NO: ) , E (SEQ ID NO: ) , F (SEQ ID NO: ) , and G (SEQ ID NO: ) between fragments and the sequences located at the junctions. The restriction sites used to generate each fragment as well as synthetic linker sequences which are used to join the fragments are described for each junction. The location of several gene coding regions and regulatory elements is also given. The following two conventions are used: numbers in parentheses, () , refer to amino acids, and restrictions sites in brackets, [] , indicate the remnants of sites which are destroyed during construction. The following abbreviations are used: swinepox virus (SPV) , pseudorabies virus (PRV) , Escherichia coli (E. coli) , pox synthetic early promoter 1 late promoter 2 (EP1LP2) , pox synthetic late promoter 1 (LPl) , base pairs (BP) .

Figures 34A-34E:

Detailed description of Swinepox Virus S-SPV-055 and the DNA insertion in Homology Vector 789-41.73. Diagram showing the orientation of DNA fragments assembled in plasmid 789-41.73. The origin of each fragment is indicated in the table. The sequences located at each of the junctions between fragments is also shown. Figures 34A-34E show the sequences located at Junction A (SEQ ID NOS: ) , (SEQ ID NO: ) , C (SEQ ID NO: ) , D (SEQ ID NO: ) , E (SEQ ID NO: ) , F (SEQ ID NO: ) , G (SEQ ID NO: ) , and H (SEQ ID NO: ) between fragments and the sequences located at the junctions. The restriction sites used to generate each fragment as well as synthetic linker sequences which are used to join the fragments are described for each junction. The location of several gene coding regions and regulatory elements is also given. The following two conventions are used: numbers in parentheses, (), refer to amino acids, and restrictions sites in brackets, [] , indicate the remnants of sites which are destroyed during construction. The following abbreviations are used: swinepox virus (SPV) , pseudorabies virus (PRV) , Escherichia coli (E. coli) , pox synthetic late promoter 2 early promoter 2 (LP2EP2) , pox synthetic early promoter 1 late promoter 2 (EP1LP2) , pox synthetic late promoter 1 (LPl) , base pairs (BP) .

PETAILEP PESCRIPTION OF THE INVENTION

The present invention provides a recombinant swinepox virus comprising a foreign DNA sequence inserted into the swinepox virus genomic DNA, wherein the foreign DNA sequence is inserted within a Hindlll K fragment of the swinepox virus genomic DNA and is capable of being expressed in a swinepox virus infected host cell.

In one embodiment the recombinant swinepox virus contains the foreign DNA sequence is inserted into an approximately 2 kB Hindlll to BamHI subfragment of the Hindlll N fragment of the swinepox virus genomic DNA. In another embodiment the foreign DNA sequence is inserted into an open reading frame within an approximately 2 kB Hindlll to BamHI subfragment of the Hindlll N fragment of the swinepox virus genomic DNA. In another embodiment the the open reading frame encodes a I7L gene.

In another embodiment the foreign DNA sequence is inserted within a EcoRV restriction endonuclease site within the approximately 2 kB Hindlll to BamHI subfragment of the swinepox virus genomic DNA. In another embodiment the foreign DNA sequence is inserted within a SnaBI restriction endonuclease site within the approximately 2.0 kB Hindlll to BamHI subfragment of the swinepox virus genomic DNA.

In another embodiment the foreign DNA sequence is inserted within an approximately 1.2 kB BamHI to Hindlll subfragment of the Hindlll N fragment of the swinepox virus genomic DNA. In another embodiment the foreign DNA sequence is inserted into an open reading frame within an approximately 1.2 kB BamHI to Hindlll subfragment of the Hindlll N fragment of the swinepox virus genomic DNA. In another embodiment the foriegn DNA sequence is inserted into an open reading frame which encodes a I4L gene. In another embodiment the foreign DNA sequence is inserted within a Bglll restriction endonuclease site within the approximately 1.2 kB BamHI to Hindlll subfragment of the swinepox virus genomic DNA.

The present invention provides a recombinant swinepox virus comprising a foreign DNA sequence inserted into the swinepox virus genomic DNA, wherein the foreign DNA sequence is inserted within a Hindlll M fragment of the swinepox virus genomic DNA and is capable of being expressed in a swinepox virus infected host cell.

In one embodiment the recombinant swinepox virus contains the foreign DNA sequence inserted into an approximately 2 kB Bglll to Hindlll subfragment of the Hindlll M fragment of the swinepox virus genomic DNA. In another embodiment the foreign DNA sequence is inserted into an open reading frame within an approximately 2 kB Bglll to Hindlll subfragment of the Hindlll M fragment of the swinepox virus genomic DNA. In another embodiment the open reading frame encodes a OIL gene. In the preferred embodiment the foreign DNA sequence is inserted within a Bglll restriction endonuclease site within the approximately 2 kB Bglll to Hindlll subfragment of the swinepox virus genomic DNA.

In another embodiment the recombinant swinepox virus contains the foreign DNA sequence inserted within an approximately 3.6 kB larger Hindlll to Bglll subfragment of the Hindlll M fragment of the swinepox virus genomic DNA. In another embodiment the foreign DNA sequence is inserted into an open reading frame within an approximately 3.6 kB larger Hindlll to Bglll subfragment of the Hindlll M fragment of the swinepox virus genomic DNA. In another embodiment the open reading frame encodes a I4L gene In one embodiment the foreign DNA sequence of the recombinant swinepox virus is inserted within a non- essential Open Reading Frame (ORF) of the Hindlll M fragment. Example of ORF's include, but are not limited to: I4L, I2L, OIL, and E10L.

In another embodiment the foreign DNA sequence of the recombinant swinepox virus is inserted within an approximately 2 Kb Hindlll to Bglll subfragment of the Hindlll M fragment of the swinepox virus genomic DNA. In a preferred embodiment the foreign DNA sequence is inserted within a Bglll site located within the approximately 2 Kb Hindlll to Bglll subfragment of the swinepox virus genomic DNA.

In another embodiment the foreign DNA sequence is inserted within a larger Hindlll to Bglll subfragment of the Hindlll M fragment of the swinepox virus genomic DNA. In a preferred embodiment the foreign DNA sequence is inserted within an AccI site located within the larger Hindlll to Bglll subfragment of the swinepox virus genomic DNA.

In another embodiment the recombinant swinepox virus further comprises a foreign DNA sequence inserted into an open reading frame encoding swinepox virus thymidine kinase. In one embodiment the foreign DNA sequence is inserted into a Ndel site located within the open reading frame encoding the swinepox virus thymidine kinase.

This invention provides a recombinant swinepox virus comprising a foreign DΝA sequence inserted into the swinepox virus genomic DΝA, wherein the foreign DΝA sequence is inserted within a Hindlll K fragment of the swinepox virus genomic DΝA and is capable of being expressed in a swinepox virus infected host cell. In one embodiment the foreign DNA sequence is inserted into an approximately 3.2 kB subfragment of the Hindlll K fragment of the swinepox virus genomic DNA. In another embodiment the foreign DNA sequence is inserted into an open reading frame within an approximately 3.2 kB subfragment of the Hindlll K fragment of the swinepox virus genomic DNA. In another embodiment the open reading frame encodes a B18R gene. In another embodiment the open reading frame encodes a B4R gene.

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 swinepox viral 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.

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. In addition, 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.

The invention further provides a foreign DNA sequence or foreign RNA which encodes a polypeptide. Preferably, the polypeptide is antigenic in the animal. 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 a recombinant swinepox virus capable of replication which contains a foreign DNA encoding a polypeptide which is a detectable marker. Preferably the detectable marker is the polypeptide E. coli ?-galactosidase or E. coli beta -glucuronidase . Preferably, the insertion site for the foreign DNA encoding E. coli -galactosidase is the AccI restriction endonuclease site located within the Hindlll M fragment of the swinepox viral DNA. 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. For purposes of this invention, a "polypeptide which is a detectable marker" includes the bimer, trimer and tetramer form of the polypeptide. E. coli β- galactosidase is a tetramer composed of four polypeptides or monomer sub-units.

The invention further provides a recombinant swinepox virus capable of replication which contains foreign DNA encoding an antigenic polypeptide which is or is from pseudorabies virus (PRV) g50 (gD) , pseudorabies virus

(PRV) gll (gB) , Pseudorabies virus (PRV) gill (gC) , pseudorabies virus (PRV) glycoprotein H, pseudorabies virus (PRV) glycoprotein E, Transmissible gastroenteritis

(TGE) glycoprotein 195, Transmissible gastroenteritis (TGE) matrix protein, swine rotavirus glycoprotein 38, swine parvovirus capsid protein, Serpul ina hydodys ente iae 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 Virus (PRV) g50 (gD) . Preferably, the antigenic protein is Newcastle Disease Virus (NDV) hemagglutinin-neuraminidase.

The invention further provides a recombinant swinepox virus capable of replication which contains foreign DNA 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 for a recombinant swinepox virus capable of replication which contains foreign DNA encoding pseudorabies virus (PRV) g50 (gD) . This recombinant swinepox virus can be further engineered to contain foreign DNA encoding a detectable marker, such as E . coli _/S-galactosidase. A preferred site within the swinepox viral genome for insertion of the foreign DNA encoding PRV g50 (gD) and E. coli β-galactosidase is the AccI site within the Hindlll M fragment of the swinepox viral DNA. 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 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 2339.

The invention further provides for a recombinant swinepox virus capable of replication which contains foreign DNA encoding pseudorabies virus (PRV) gill (gC) . This recombinant swinepox virus can also be further engineered to contain foreign DNA encoding a detectable marker, such as E. coli jβ-galactosidase. A preferred site within the swinepox viral DNA for insertion of the foreign DNA encoding PRV C gene and E. coli _/S-galactosidase is the AccI site within the Hindlll M fragment of the swinepox viral DNA. Preferably, this recombinant swinepox virus is designated S-SPV-011, S-SPV-012, or S-SPV-013. The swinepox virus designated S-SPV-013 has been deposited on July 16, 1993 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 2418.

The invention further provides for a recombinant swinepox virus capable of replication which contains foreign DNA encoding pseudorabies virus (PRV) gll (gB) . This recombinant swinepox virus can also be further engineered to contain foreign DNA encoding a detectable marker, such as E . coli β-galactosidase . A preferred site within the swinepox viral DNA for insertion of the foreign DNA encoding PRV gll (gB) and E. coli β-galactosidase is the AccI site within the Hindlll M fragment of the swinepox viral DNA. Preferably, this recombinant swinepox virus is designated S-SPV-015 (ATCC Accession No. VR 2466) . The S-SPV-015 swinepox virus has been deposited on July 22, 1994 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 2466.

The invention further provides for a recombinant swinepox virus capable of replication which contains foreign DNA encoding pseudorabies virus (PRV) g50 (gD) and foreign DNA encoding pseudorabies virus (PRV) gill (gC) . This recombinant swinepox virus can also be further engineered to contain foreign DNA encoding a detectable marker, such as E. coli S-galactosidase. A preferred site within the swinepox viral DNA for insertion of the foreign DNA encoding PRV g50 (gD) , PRV gill (gC) and E. coli β- galactosidase is the AccI site within the Hindlll M fragment of the swinepox viral DNA.

The invention further provides for a recombinant swinepox virus capable of replication which contains foreign DNA encoding pseudorabies virus (PRV) g50 (gD) and foreign DNA encoding pseudorabies virus (PRV) gll (gB) . This recombinant swinepox virus can also be further engineered to contain foreign DNA encoding a detectable marker, such as E. coli jβ-galactosidase. A preferred site within the swinepox viral genome for insertion of foreign DNA encoding PRV g50 (gD) , PRV gll (gB) and E. coli β- galactosidase is the AccI site within the Hindlll M fragment of the swinepox viral DNA. The invention further provides for a recombinant swinepox virus capable of replication which contains foreign DNA encoding pseudorabies virus (PRV) gill (gC) and foreign DNA encoding pseudorabies virus (PRV) gll (gB) . This recombinant swinepox virus can also be further engineered to contain foreign DNA encoding a detectable marker, such as E. coli 0-galactosidase. A preferred site within the swinepox viral genome for insertion of foreign DNA encoding PRV gill (gC) , PRV gll (gB) and E. coli β- galactosidase is the AccI site within the Hindlll M fragment of the swinepox viral DNA.

The invention further provides for a recombinant swinepox virus capable of replication which contains foreign DNA encoding pseudorabies virus (PRV) g50 (gD) , foreign DNA encoding pseudorabies virus (PRV) gill (gC) , and foreign DNA encoding pseudorabies virus (PRV) gll (gB) . This recombinant swinepox virus can also be further engineered to contain foreign DNA encoding a detectable marker, such as E. coli 3-galactosidase.

A preferred site within the swinepox viral genome for insertion of foreign DNA encoding PRV g50 (gD) , PRV gill

(gC) , PRV gll (gB) and E. coli -galactosidase is the AccI site within the Hindlll M fragment of the swinepox viral DNA.

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 present invention further provides a recombinant swinepox virus which comprises a foreign DNA sequence inserted into a non-essential site of the swinepox genome, wherein the foreign DNA sequence encodes an antigenic polypeptide derived from infectious bovine rhinotracheitis virus and is capable of being expressed in a host infected by the recombinant swinepox virus. Examples of such antigenic polypeptide are infectious bovine rhinotracheitis virus glycoprotein E and glycoprotein G. Preferred embodiment of this invention are recombinant swinepox viruses designated S-SPV-017 and S-SPV-019.

The present invention further provides a recombinant swinepox virus which comprises a foreign DNA sequence inserted into a non-essential site of the swinepox genome, wherein the foreign DNA sequence encodes an antigenic polypeptide derived from infectious laryngotracheitis virus and is capable of being expressed in a host infected by the recombinant swinepox virus.

Examples of such antigenic polypeptide are infectious laryngotracheitis virus glycoprotein G and glycoprotein

I. Preferred embodiment of this invention are recombinant swinepox viruses designated S-SPV-014 and S- SPV-016.

In one embodiment of the recombinant swinepox virus the foreign DNA sequence encodes a cytokine. In another embodiment the cytokine is chicken myelomonocytic growth factor (cMGF) or chicken interferon (cIFN) . Cytokines include, but are not limited to: transforming growth factor beta, epidermal growth factor family, fibroblast growth factors, hepatocyte growth factor, insulin-like growth factor, vascular endothelial growth factor, interleukin 1, IL-1 receptor antagonist, interleukin-2, interleukin-3, interleukin-4, interleukin-5, interleukin- 6, IL-6 soluble receptor, interleukin-7, interleukin-8, interleukin- 9 , interleukin-10 , interleukin-11 , interleukin-12, interleukin-13 , angiogenin, chemokines, colony stimulating factors, granulocyte-macrophage colony stimulating factors, erythropoietin, interferon, interferon gamma, c-kit ligand, leukemia inhibitory factor, oncostatin M, pleiotrophin, secretory leukocyte protease inhibitor, stem cell factor, tumor necrosis factors, and soluble TNF receptors. These cytokines are from humans, bovine, equine, feline, canine, porcine or avian. Preferred embodiments of such recombinant virus are designated S-SPV-042, and S-SPV-043.

The present invention further provides a recombinant swinepox virus which comprises a foreign DNA sequence inserted into a non-essential site of the swinepox genome, wherein the foreign DNA sequence encodes an antigenic polypeptide derived from a human pathogen and is capable of being expressed in a host infected by the recombinant swinepox virus .

Recombinant SPV expressing cytokines is used to enhance the immune response either alone or when combined with vaccines containing cytokines or antigen genes of disease causing microorganisms.

Antigenic polypeptide of a human pathogen which are derived from human herpesvirus include, but are not limited to: hepatitis B virus and hepatitis C virus hepatitis B virus surface and core antigens, hepatitis C virus, human immunodeficiency virus, herpes simplex virus-1, herpes simplex virus-2, human cytomegalovirus, Epstein-Barr virus, Varicella-Zoster virus, human herpesvirus-6, human herpesvirus-7, human influenza, measles virus, hantaan virus, pneumonia virus, rhinovirus, poliovirus, human respiratory syncytial virus, retrovirus, human T-cell leukemia virus, rabies virus, mumps virus, malaria (Plas/nodiiuπ falciparum) , Bordetella pertussis, Diptheria, Rickettsia prowazekii , Borrelia berfdorferi , Tetanus toxoid, malignant tumor antigens.

In one embodiment of the invention, a recombinant swinepox virus contains the foreign DNA sequence encoding hepatitis B virus core protein. Preferably, such virus recombinant virus is designated S-SPV-031.

The present invention further provides a recombinant swinepox virus which comprises a foreign DNA sequence inserted into a non-essential site of the swinepox genome, wherein the foreign DNA sequence encodes a cytokine capable of stimulating an immune in a host infected by the recombinant swinepox virus and is capable of being expressed in the host infected.

In one embodiment of the invention, a recombinant swinepox virus contains a foreign DNA sequence encoding human interleukin-2. Preferably, such recombinant virus is designated S-SPV-035.

The present invention further provides a recombinant swinepox virus which comprises a foreign DNA sequence inserted into a non-essential site of the swinepox genome, wherein the foreign DNA sequence encodes an antigenic polypeptide derived from an equine pathogen and is capable of being expressed in a host infected by the recombinant swinepox virus .

The antigenic polypeptide of an equine pathogen can derived from equine influenza virus, or equine herpesvirus . In one embodiment the antigenic polypeptide is equine influenza neuraminidase or hemagglutinin. Examples of such antigenic polypeptide are equine influenza virus type A/Alaska 91 neuraminidase, equine influenza virus type A/Prague 56 neuraminidase, equine influenza virus type A/Miami 63 neuraminidase, equine influenza virus type A/Kentucky 81 neuraminidase, equine influenza virus type A/Kentucky 92 neuraminidase equine herpesvirus type 1 glycoprotein B, equine herpesvirus type 1 glycoprotein D, Streptococcus equi , equine infectious anemia virus, equine encephalitis virus, equine rhinovirus and equine rotavirus. Preferred embodiments of such recombinant virus are designated S- SPV-033, S-SPV-034, S-SPV-038, S-SPV-039 and S-SPV-041.

The present invention further provides an antigenic polypeptide which includes, but is not limited to: hog cholera virus gEl, hog cholera virus gE2, swine influenza virus hemagglutinin, neurominidase, matrix and nucleoprotein, pseudorabies virus gB, gC and gD, and PRRS virus ORF7.

The present invention further provides a recombinant swinepox virus which comprises a foreign DNA sequence inserted into a non-essential site of the swinepox genome, wherein the foreign DNA sequence encodes an antigenic polypeptide derived from bovine respiratory syncytial virus or bovine parainfluenza virus, and is capable of being expressed in a host infected by the recombinant swinepox virus.

For example, the antigenic polypeptide of derived from infectious bovine rhinotracheitis virus gE, bovine respiratory syncytial virus equine pathogen can derived from equine influenza virus is bovine respiratory syncytial virus attachment protein (BRSV G) , bovine respiratory syncytial virus fusion protein (BRSV F) , bovine respiratory syncytial virus nucleocapsid protein

(BRSV N) , bovine parainfluenza virus type 3 fusion protein, and the bovine parainfluenza virus type 3 hemagglutinin neuraminidase. In a preferred embodiment the recombinant swinepox virus is designated S-SPV-045.

Preferred embodiments of a recombinant virus containing a foreign DNA encoding an antigenic polypeptide from a bovine respiratory syncytial virus are designated S-SPV- 020, S-SPV-029, and S-SPV-030. And a preferred embodiment of a recombinant virus containing a foreign DNA encoding an antigenic polypeptide from a bovine parainfluenza virus are designated S-SPV-028.

The present invention further provides a recombinant swinepox virus which comprises a foreign DNA sequence inserted into a non-essential site of the swinepox genome, wherein the foreign DNA sequence encodes bovine viral diarrhea virus (BVDV) glycoprotein 48 or glycoprotein 53, and wherein the foreign DNA sequence is capable of being expressed in a host infected by the recombinant swinepox virus. Preferred embodiments of such virus are designated S-SPV-032, S-SPV-040, S-SPV- 049, and S-SPV-050.

The present invention further provides a recombinant swinepox virus which comprises a foreign DNA sequence inserted into a non-essential site of the swinepox genome, wherein the foreign DNA sequence encodes an antigenic polypeptide derived from infectious bursal disease virus and wherein the foreign DNA sequence is capable of being expressed in a host infected by the recombinant swinepox virus. Examples of such antigenic polypeptide are infectious bursal disease virus polyprotein and VP2. Preferred embodiments of such virus are designated S-SPV-026 and S-SPV-027. The present invention further provides a recombinant swinepox virus in which the foreign DNA sequence encodes an antigenic polypeptide which includes, but is not limited to: MDV gA, MDV gB, MDV gD, NDV HN, NDV F, ILT gB, ILT gl, ILT gD, IBDV VP2, IBDV VP3, IBDV VP4 , IBDV polyprotein, IBV spike, IBV matrix, avian encephalomyelitis virus, avian reovirus, avian paramyxovirus, avian influenza virus, avian adenovirus, fowl pox virus, avian coronavirus, avian rotavirus, chick anemia virus, Salmonella spp . E. coli , Pasteurella spp . , Bordetella spp . , Eimeria spp . , Histomonas spp . , Trichomonas spp. , Poultry nematodes, cestodes, trematodes, poultry mites/lice, and poultry protozoa.

The invention further provides that the inserted foreign DNA sequence is under the control of a promoter. In one embodiment the is a swinepox viral promoter. In another embodiment the foreign DNA sequence is under control of an endogenous upstream poxvirus promoter. In another embodiment the foreign DNA sequence is under control of a heterologous upstream promoter.

For purposes of this invention, promoters include but is not limited to: synthetic pox viral promoter, pox synthetic late promoter 1, pox synthetic late promoter 2 early promoter 2, pox OIL promoter, pox I4L promoter, pox I3L promoter, pox I2L promoter, pox I1L promoter, pox E10R promoter, PRV gX, HSV-1 alpha 4, HCMV immediate early, MDV gA, MDV gB, MDV gD, ILT gB, BHV-1.1 VP8 and ILT gD. Alternate promoters are 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.

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 essentially of a double-stranded foreign DNA sequence or (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 another embodiment of the present invention, the double-stranded swinepox viral DNA of the homology vectors described above is homologous to genomic DNA present within the Hindlll M fragment. In another embodiment the double-stranded swinepox viral DNA of the homology vectors described above is homologous to genomic DNA present within an approximately 2 Kb Hindlll to Bglll sub-fragment. In a preferred embodiment the double- stranded swinepox viral DNA is homologous to genomic DNA present within the Bgll l site located in this Hindlll to Bglll subfragment.

In another embodiment the double-stranded swinepox viral DNA is homologous to genomic DNA present within the open reading frame contained in the larger Hindlll to Bglll subfragment. Preferably, the double-stranded swinepox viral DNA is homologous to genomic DNA present within the AccI restriction endonuclease site located in the larger Hindlll to Bglll subfragment.

In a preferred embodiment the homology vectors are designated 752-29.33, 751-07.Al, 751-56.Al, 751-22.1, 746-94.1, 767-67.3, 738-94.4, and 771-55.11. 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, the polypeptide is antigenic in the animal. Preferably, the antigenic polypeptide is or is from pseudorabies virus (PRV) g50 (gD) , pseudorabies virus (PRV) gll (gB) , Pseudorabies virus (PRV) gill (gC) , 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 53 and g48, 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 gEl and gE2, Swine Influenza Virus, African Swine Fever Virus or Mycoplasma hyopneumoniae , swine influenza virus hemagglutinin, neuraminidase and matrix and nucleoprotein, PRRS virus ORF7, and hepatitis B virus core protein.

In an embodiment of the present invention, the double stranded foreign DNA sequence in the homology vector encodes an antigenic polypeptide derived from a human pathogen.

For example, the antigenic polypeptide of a human pathogen is derived from human herpesvirus, herpes simplex virus-1, herpes simplex virus-2, human cytomegalovirus, Epstein-Barr virus, Varicell-Zoster virus, human herpesvirus-6, human herpesvirus-7, human influenza, human immunodeficiency virus, rabies virus, measles virus, hepatitis B virus and hepatitis C virus. Furthermore, the antigenic polypeptide of a human pathogen may be associated with malaria or malignant tumor from the group conisting of Plasmodium fal ciparum, Bordetella pertusis, and malignant tumor.

In an embodiment of the present invention, the double stranded foreign DNA sequence in the homology vector encodes a cytokine capable of stimulating human immune response. In one embodiment the cytokine is a chicken myelomonocytic growth factor (cMGF) or chicken interferon (cIFN) . For example, the cytokine can be, but not limited to, interleukin-2, interleukin-6, interleukin-12, interferons, granulocyte-macrophage colony stimulating factors, and interleukin receptors.

In an embodiment of the present invention, the double stranded foreign DNA sequence in the homology vector encodes an antigenic polypeptide derived from an equine pathogen.

The antigenic polypeptide of an equine pathogen can derived from equine influenza virus or equine herpesvirus. Examples of such antigenic polypeptide are equine influenza virus type A/Alaska 91 neuraminidase, equine influenza virus type A/Prague 56 neuraminidase, equine influenza virus type A/Miami 63 neuraminidase, equine influenza virus type A/Kentucky 81 neuraminidaseequine herpesvirus type 1 glycoprotein B, and equine herpesvirus type 1 glycoprotein D.

In an embodiment of the present invention, the double stranded foreign DNA sequence of the homology vector encodes an antigenic polypeptide derived from bovine respiratory syncytial virus or bovine parainfluenza virus.

For example, the antigenic polypeptide is derived from infectious bovine rhinotracheitis gE, bovine respiratory syncytial virus attachment protein (BRSV G) , bovine respiratory syncytial virus fusion protein (BRSV F) , bovine respiratory syncytial virus nucleocapsid protein (BRSV N) , bovine parainfluenza virus type 3 fusion protein, and the bovine parainfluenza virus type 3 hemagglutinin neuraminidase.

In an embodiment of the present invention, the double stranded foreign DNA sequence of the homology vector encodes an antigenic polypeptide derived from infectious bursal disease virus. Examples of such antigenic polypeptide are infectious bursal disease virus polyprotein and infectious bursal disease virus VP2, VP3 , or VP4.

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 of the homology vectors described above is homologous to genomic DNA present within the open reading frame encoding swinepox thymidine kinase. Preferably, the double-stranded swinepox viral DNA is homologous to genomic DNA present within the Ndel restriction endonuclease site located in the open reading frame encoding swinepox thymidine kinase.

The invention further provides a homology vectors described above, the foreign DNA sequence of which is under control of a promoter located upstream of the foreign DNA sequence. The promoter can be an endogenous swinepox viral promoter or an exogenous promoter. Promoters include, but are not limited to: synthetic pox viral promoter, pox synthetic late promoter 1, pox synthetic late promoter 2 early promoter 2, pox OIL promoter, pox I4L promoter, pox I3L promoter, pox I2L promoter, pox I1L promoter, pox E10R promoter, PRV gX, HSV-1 alpha 4, HCMV immediate early, BHV-1.1 VP8, infectious laryngotracheitis virus glycoprotein B, infectious laryngotracheitis virus gD, marek's disease virus glycoprotein A, marek's disease virus glycoprotein B, and marek's disease virus glycoprotein D.

This invention provides a recombinant swinpox virus designated S-SPV-044, S-SPV-046, S-SPV-047, S-SPV-048, S- SPV-052, S-SPV-051, S-SPV-053, S-SPV-054, S-SPV-055, S- SPV-056, S-SPV-057, S-SPV-058, S-SPV-059, S-SPV-060, S- SPV-061, and S-SPV-062.

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 swinepox 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 10³ to 10⁹ PFU/dose.

The present invention also provides a method of immunizing an animal, wherein the animal is a human, 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 hyody s ente i ae , bovine viral diarrhea, Newcastle disease, swine influenza, PRRS, bovine respiratory synctial virus, bovine parainfluenza virus type 3, foot and mouth disease, hog cholera, African swine fever or Mycoplasma hyopneumoniae . For purposes of this invention, the method of immunizing also includes immunizing the animal against human pathogens, bovine pathogens, equine pathogens, avian pathogens described in the preceding part of this section.

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 provides a recombinant SPV which when inserted with a foreign DNA sequence or gene may be employed as a diagnostic assay. In one embodiment FIV env and gag genes and D. immi tis p39 and 22kd are employed in a diagnostic assay to detect feline immunodeficiency caused by FIV and to detect heartworm caused by D. immi ts, respectively.

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 ESK-4 cell, PK-15 cell or 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 viruses described above are detailed below in Materials and Methods.

EXPERIMENTAL DETAILS

Materials and Methods

PREPARATION OF SWINEPOX VIRUS STOCK SAMPLES. Swinepox virus (SPV) samples were prepared by infecting embryonic swine kidney (EMSK) cells, ESK-4 cells, PK-15 cells or Vero 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% C0₂ 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 10⁶ PFU/ml .

PREPARATION OF SPV DNA. For swinepox virus DNA isolation, a confluent monolayer of EMSK cells in a T175 cm² 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 Na₂HP0₄, 0.2g KH₂P0₄, 0.8g NaCL and 0.2g KC1 per liter H₂0) (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, ESK-4, PK-15 or Vero for SPV or VERO for PRV) in a 25 cm² 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% _/S-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: 6.61g Tris-HCl, 0.97g Tris-base, 9. Og NaCl and 2. Og Sodium Azide per liter H₂0) . 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 ³⁵S-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 primers 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% C0₂ 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 H₂0. Separately, 50 micrograms of Lipofectin Reagent was diluted to 100 μl with H₂0. 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% C0₂ 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% C0₂ 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 j8-galactosidase (BLUOGAL AND CPRG ASSAYS) . When the E. coli β- galactosidase (lacZ) marker gene was incorporated into a reco binant 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 agarose overlay during the plaque assay, and plaques 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 g50 (gD) 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 (PRV g50 (gD) ,- 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% H₂0₂ in PBS) . Plaques expressing the correct antigen stain black.

PROCEDURE FOR PURIFICATION OF VIRAL GLYCOPROTEINS FOR USE AS DIAGNOSTICS. Viral glycoproteins are purified using antibody affinity columns. To produce monoclonal antibodies, 8 to 10 week old BALB/c female mice are vaccinated intraperitoneally seven times at two to four week intervals with 10⁷ PFU of S-SPV-009, -014, -016, - 017, -018, or -019. Three weeks after the last vaccination, mice are injected intraperitoneally with 40 mg of the corresponding viral glycoprotein. Spleens are removed from the mice three days after the last antigen dose.

Splenocytes are fused with mouse NSl/Ag4 plasmacytoma cells by the procedure modified from Oi and Herzenberg,

(41) . Splenocytes and plasmacytoma cells are pelleted together by centrifugation at 300 x g for 10 minutes.

One ml of a 50% solution of polyethylene glycol (m.w. 1300-1600) is added to the cell pellet with stirring over one minute. Dulbecco's modified Eagles's medium (5ml) is added to the cells over three minutes. Cells are pelleted by centrifugation at 300 x g for 10 minutes and resuspended in medium with 10% fetal bovine serum and containing 100 mM hypoxanthine, 0. mM aminopterin and 16 mM thymidine (HAT) . Cells (100 ml) are added to the wells of eight to ten 96-well tissue culture plates containing 100 ml of normal spleen feeder layer cells and incubated at 37°C. Cells are fed with fresh HAT medium every three to four days.

Hybridoma culture supernatants are tested by the ELISA ASSAY in 96-well microtiter plates coated with 100 ng of viral glycoprotein. Supernatants from reactive hybridomas are further analyzed by black-plaque assay and by Western Blot. Selected hybridomas are cloned twice by limiting dilution. Ascetic fluid is produced by intraperitoneal injection of 5 x 10⁶ hybridoma cells into pristane-treated BALB/c mice.

Cell lysates from S-SPV-009, -014, -016, -017, -018, or - 019 are obtained as described in PREPARATION OF INFECTED

CELL LYSATES. The glycoprotein-containing cell lysates

(100 mis) are passed through a 2-ml agarose affinity resin to which 20 mg of glycoprotein monoclonal antibody has been immobilized according to manufacturer's instructions (AFC Medium, New Brunswick Scientific,

Edison, N.J.) . The column is washed with 100 ml of 0.1%

Nonidet P-40 in phosphate-buffered saline (PBS) to remove nonspecifically bound material. Bound glycoprotein is eluted with 100 mM carbonate buffer, pH 10.6 (40) . Pre- and posteluted fractions are monitored for purity by reactivity to the SPV monoclonal antibodies in an ELISA system.

ELISA ASSAY. A standard enzyme-linked immunosorbent assay (ELISA) protocol is used to determine the immune status of cattle following vaccination and challenge.

A glycoprotein antigen solution (100 ml at ng/ml in PBS) is allowed to absorb to the wells of microtiter dishes for 18 hours at 4°C. The coated wells are rinsed one time with PBS. Wells are blocked by adding 250 ml of PBS containing 1% BSA (Sigma) and incubating 1 hour at 37°C. The blocked wells are rinsed one time with PBS containing 0.02% Tween 20. 50 ml of test serum (previously diluted 1:2 in PBS containing 1% BSA) are added to the wells and incubated 1 hour at 37°C. The antiserum is removed and the wells are washed 3 times with PBS containing 0.02% Tween 20. 50 ml of a solution containing anti-bovine IgG coupled to horseradish peroxidase (diluted 1:500 in PBS containing 1% BSA, Kirkegaard and Perry Laboratories, Inc.) is added to visualize the wells containing antibody against the specific antigen. The solution is incubated 1 hour at 37°C, then removed and the wells are washed 3 times with PBS containing 0.02% Tween 20. 100 ml of substrate solution (ATBS, Kirkegaard and Perry Laboratories, Inc.) are added to each well and color is allowed to develop for 15 minutes. The reaction is terminated by addition of 0.1M oxalic acid. The color is read at absorbance 410nm on an automatic plate reader.

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. Three promoter cassettes LPl, EP1 and LP2 based on promoters that have been defined in the vaccinia virus (1, 7 and 8) were designed. 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) . VACCINATION STUDIES IN SWINE USING RECOMBINANT SWINEPOX VIRUS CONTAINING PSEUDORABIES VIRUS GLYCOPROTEIN GENES:

Young weaned pigs from pseudorabies-free herd are used to test the efficacy of the recombinant swinepox virus containing one or more of the pseudorabies virus glycoprotein genes (SPV/PRV) . The piglets are inoculated intramuscularly, intradermally or orally about 10³ to 10⁷ plaque forming units (PFU) of the recombinant SPV/PRV viruses.

Immunity is determined by measuring PRV serum antibody levels and by challenging the vaccinated pigs with virulent strain of pseudorabies virus. Three to four weeks post-vaccination, both vaccinated and non- vaccinated groups of pigs are challenged with virulent strain of pseudorabies virus (VDL4892) . Post challenge, the pigs are observed daily for 14 days for clinical signs of pseudorabies.

Serum samples are obtained at the time of vaccination, challenge, and at weekly intervals for two to three weeks post-vaccination and assayed for serum neutralizing antibody.

CLONING OF EQUINE INFLUENZA VIRUS HEMAGGLUTININ AND

NEURAMINIDASE GENES. The equine influenza virus hemagglutinin (HA) and Neuraminidase (NA) genes was cloned essentially as described by Katz et al . (42) for the HA gene of human influenza virus. Viral RNA was prepared from virus grown in MDBK cells (for Influenza

A/equine/Alaska/91 and Influenza A/equine/Miami/63) and MDCK cells (for Influenza A/equine/Prague/56 and Influenza A/equine/Kentucky/81) was first converted to cDNA utilizing an oligo nucleotide primer specific for the target gene. The cDNA was used as a template for PCR cloning (51) of the targeted gene region. The PCR primers were designed to incorporate restriction sites which permit the cloning of the amplified coding regions into vectors containing the appropriate signals for expression in EHV. One pair of oligo nucleotide primers was required for each coding region. The HA gene coding regions from the serotype 2 (H3) viruses (Influenza A/equine/Miami/63 , Influenza A/equine/Kentucky/81, and Influenza A/equine/Alaska/91) was cloned utilizing the f o l l o w i n g p r i m e r s 5 ' GGAGGCCTTCATGACAGACAACCATTATTTTGATACTACTGA-3' (SEQ ID NO: 120) for cDNA priming and combined with 5'- GAAGGCCTTCTCAAATGCAAATGTTGCATCTGATGTTGCC-3' (SEQ ID NO: 121) for PCR. The HA gene coding region from the serotype 1 (H7) virus (Influenza A/equine/Prague/56) was be cloned utilizing the following primers 5 ' - GGGATCCATGAACACTCAAATTCTAATATTAG-3' (SEQ ID NO: 122) for cDNA priming and combined with 5 ' - GGGATCCTTATATACAAATAGTGCACCGCA-3' (SEQ ID NO: 123) for PCR. The NA gene coding regions from the serotype 2 (N8) viruses (Influenza A/equine/Miami/63 , Influenza A/equine/Kentucky/81, and Influenza A/equine/Alaska/91) was cloned utilizing the following primers 5'- GGGTCGACATGAATCCAAATCAAAAGATAA-3' (SEQ ID NO: 124) for cDNA priming and combined with 5 ' - GGGTCGACTTACATCTTATCGATGTCAAA-3' (SEQ ID NO: 125) for PCR. The NA gene coding region from the serotype 1 (N7) virus (Influenza/A/equine/Prague/56) was cloned utilizing the following primers 5' -GGGATCCATGAATCCTAATCAAAAACTCTTT- 3' (SEQ ID NO: 118) for cDNA priming and combined with 5' -GGGATCCTTACGAAAAGTATTTAATTTGTGC-3' (SEQ ID NO: 119) for PCR. Note that this general strategy was used to clone the coding regions of HA and NA genes from other strains of equine influenza A virus. The EIV HA or NA genes were cloned as a blunt ended Sail or BamHI fragment into a blunt ended EcoRI site behind the LP2EP2 promoter of the SPV homology vector. CLONING OF PARAINFLUENZA-3 VIRUS FUSION AND HEMAGGLUTININ GENES. The parainfluenza-3 virus fusion (F) and hemagglutinin (HN) genes were cloned by a PCR CLONING procedure essentially as described by Katz et al. (42) for the HA gene of human influenza. Viral RNA prepared from bovine PI-3 virus grown in Madin-Darby bovine kidney (MDBK) cells was first converted to cDNA utilizing an oligonucleotide primer specific for the target gene. The cDNA was then used as a template for polymerase chain reaction (PCR) cloning (15) of the targeted region. The PCR primers were designed to incorporate restriction sites which permit the cloning of the amplified coding regions into vectors containing the appropriate signals for expression in SPV. One pair of oligonucleotides were required for each coding region. The F gene coding region from the PI-3 strain SF-4 (VR-281) was cloned using the followingprimers: 5' -TTATGGATCCTGCTGCTGTGTTGAACAACTTTGT- 3' (SEQ ID NO: 130) for cDNA priming and combined with 5' -CCGCGGATCCCATGACCATCACAACCATAATCATAGCC-3' (SEQ ID NO: 131) for PCR. The HN gene coding region from PI-3 strain SF-4 (VR-281) was cloned utilizing the following primers: 5' -CGTCGGATCCCTTAGCTGCAGTTTTTTGGAACTTCTGTTTTGA-3 ' (SEQ ID NO: 132) for cDNA priming and combined with 5'- CATAGGATCCCATGGAATATTGGAAACACACAAACAGCAC-3' (SEQ ID NO: 133) for PCR. Note that this general strategy is used to clone the coding region of F and HN genes from other strains of PI-3. The DNA fragment for PI-3 HN or F was digested with BamHI to yield an 1730 bp or 1620 bp fragment, respectively. The PI-3 HN fragment is cloned into the BamHI site next to the LP2EP2 promoter of the SPV homology vector. The PI-3 F fragment was cloned into the BamHI site next to the LP2EP2 promoter of the SPV homology vector to yield homology vector 713-55.10.

CLONING OF BOVINE VIRAL DIARRHEA VIRUS g48 and g53 GENES.

The bovine viral diarrhea g48 and g53 genes were cloned by a PCR CLONING procedure essentially as described by Katz et al . (42) for the HA gene of human influenza. Viral RNA prepared from BVD virus Singer strain grown in Madin-Darby bovine kidney (MDBK) cells was first converted to cDNA utilizing an oligonucleotide primer specific for the target gene. The cDNA was then used as a template for polymerase chain reaction (PCR) cloning (15) of the targeted region. The PCR primers were designed to incorporate restriction sites which permit the cloning of the amplified coding regions into vectors containing the appropriate signals for expression in SPV. One pair of oligonucleotides were required for each coding region. The g48 gene coding region from the BVDV Singer strain (49) was cloned using the following primers : 5' -ACGTCGGATCCCTTACCAAACCACGTCTTACTCTTGTTTTCC-3' (SEQ ID NO: 134) for cDNA priming and combined with 5'- ACATAGGATCCCATGGGAGAAAACATAACACAGTGGAACC-3' (SEQ ID NO: 135) for PCR. The g53 gene coding region from the BVDV Singer strain (49) was cloned using the following primers: 5' -CGTGGATCCTCAATTACAAGAGGTATCGTCTAC-3 ' (SEQ ID NO: 136) for cDNA priming and combined with 5'- CATAGATCTTGTGGTGCTGTCCGACTTCGCA-3' (SEQ ID NO: 137) for PCR. Note that this general strategy is used to clone the coding region of g48 and g53 genes from other strains of BVDV. The DNA fragment for BVDV g 48 was digested with BamHI to yield an 678 bp fragment. The DNA fragment for BVDV g53 was digested with Bglll and BamHI to yield an 1187 bp fragment. The BVDV g48 or g53 DNA fragments were cloned into the BamHI site next to the LP2EP2 promoter of the SPV homology vector to yield homology vectors, 727- 78.1 and 738-96, respectively.

CLONING OF BOVINE RESPIRATORY SYNCYTIAL VIRUS FUSION, NUCLEOCAPSID AND GLYCOPROTEIN GENES. The bovine respiratory syncytial virus fusion (F) , nucleocapsid (N) , and glycoprotein (G) genes were cloned by a PCR CLONING procedure essentially as described by Katz et al . (42) for the HA gene of human influenza. Viral RNA prepared from BRSV virus grown in bovine nasal turbinate (BT) cells was first converted to cDNA utilizing an oligonucleotide primer specific for the target gene. The cDNA was then used as a template for polymerase chain reaction (PCR) cloning (15) of the targeted region. The PCR primers were designed to incorporate restriction sites which permit the cloning of the amplified coding regions into vectors containing the appropriate signals for expression in SPV. One pair of oligonucleotides were required for each coding region. The F gene coding region from the BRSV strain 375 (VR-1339) was cloned using the f o l l o w i n g p r i m e r s : 5 ' - TGCAGGATCCTCATTTACTAAAGGAAAGATTGTTGAT-3' (SEQ ID NO: 138) for cDNA priming and combined with 5' - CTCTGGATCCTACAGCCATGAGGATGATCATCAGC-3' (SEQ ID NO: 139) for PCR. The N gene coding region from BRSV strain 375 (VR-1339) was cloned utilizing the following primers: 5'- CGTCGGATCCCTCACAGTTCCACATCATTGTCTTTGGGAT-3' (SEQ ID NO: 140) for cDNA priming and combined with 5'- CTTAGGATCCCATGGCTCTTAGCAAGGTCAAACTAAATGAC-3' (SEQ ID NO: 141) for PCR. The G gene coding region from BRSV strain 375 (VR-1339) was cloned utilizing the following primers: 5' -CGTTGGATCCCTAGATCTGTGTAGTTGATTGATTTGTGTGA-3' (SEQ ID NO: 142) for cDNA priming and combined with 5'- CTCTGGATCCTCATACCCATCATCTTAAATTCAAGACATTA-3' (SEQ ID NO: 143) for PCR. Note that this general strategy is used to clone the coding region of F, N and G genes from other strains of BRSV. The DNA fragments for BRSV F, N, or G were digested with BamHI to yield 1722 bp, 1173 bp, or 771 bp fragments, respectively. The BRSV F, N, and G DNA fragments were cloned into the BamHI site next to the LP2EP2 promoter of the SPV homology vector to yield homology vectors, 727-20.10, 713-55.37 and 727-20.5, respectively.

RNA ISOLATED FROM CONCANAVALIN A STIMULATED CHICKEN SPLEEN CELLS. Chicken spleens were dissected from 3 week old SPAFAS hatched chicks, washed, and disrupted through a syringe/needle to release cells. After allowing stroma and debri to settle out, the cells were pelleted and washed twice with PBS. The cell pellet was treated with a hypotonic lysis buffer to lyse red blood cells, and splenocytes were recovered and washed twice with PBS. Splenocytes were resuspended at 5 x 10⁶ cells/ml in RPMI containing 5% FBS and 5 μg/ml Concanavalin A and incubated at 39° for 48 hours. Total RNA was isolated from the cells using guanidine isothionate lysis reagents and protocols from the Promega RNA isolation kit (Promega Corporation, Madison Wl) . 4μg of total RNA was used in each 1st strand reaction containing the appropriate antisense primers and AMV reverse transcriptase (Promega Corporation, Madison Wl) . cDNA synthesis was performed in the same tube following the reverse transcriptase reaction, using the appropriate sense primers and Vent^® DNA polymerase (Life Technologies, Inc. Bethesda, MD) .

SCREEN FOR RECOMBINANT HERPESVIRUS EXPRESSING ENZYMATIC MARKER GENES. When the E.coli -glucuronidase (uidA) marker gene was incorporated into a recombinant virus the plaques containing recombinants were visualized by a simple assay. The enzymatic substrate was incorporated (300 μg/ml) into the agarose overlay during the plaque assay. For the uidA marker gene the substrate X-Glucuro Chx (5-bromo-4-chloro-3-indolyl-|S-D-glucuronic acid Cyclohexylammonium salt, Biosynth AG) was used. Plaques that expressed active marker enzyme turned blue. The blue plaques were then picked onto fresh ESK-4 cells and purified by further blue plaque isolation. In recombinant virus strategies in which the enzymatic marker gene is removed the assay involves plaque purifying white plaques from a background of parental blue plaques. In both cases viruses were typically purified with three rounds of plaque purification. 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 Figures 4A-4D. 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 3628 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 jβ-galactosidase (lacZ) marker gene flanked by SPV DNA. Upstream of the marker gene is an approximately 2149 base pair fragment of SPV DNA. Downstream of the marker gene is an approximately 1484 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 Figures 4A-4D. 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 Figures 4A-4D. The plasmid vector was derived from an approximately 2972 base pair Hindlll to BamHI restriction fragment of pSP64 (Promega) . Fragment 1 is an approximately 2149 base pair Hindlll to AccI restriction sub-fragment of the SPV Hindlll restriction fragment M (23) . Fragment 2 is an approximately 3006 base pair BamHI to PvuII restriction fragment of plasmid pJF751 (11) . Fragment 3 is an approximately 1484 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 /3-galactosidase (lacZ) marker gene and the PRV g50 (gD) gene flanked by SPV DNA. Upstream of the foreign genes is an approximately 2149 base pair fragment of SPV DNA. Downstream of the foreign genes is an approximately 1484 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 _/3-galactosidase (lacZ) marker gene is under the control of a synthetic late pox promoter (LPl) and the g50 (gD) gene is under the control of a synthetic early/late pox promoter

(EP1LP2) . A detailed description of the plasmid is given in Figures 5A-5D. 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 Figures 5A- 5D. The plasmid vector was derived from an approximately 2972 base pair Hindlll to BamHI restriction fragment of pSP64 (Promega) . Fragment 1 is an approximately 2149 base pair Hindlll to AccI restriction sub-fragment of the SPV Hindlll restriction fragment M (23) . Fragment 2 is an approximately 3006 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'- CGCGAATTCGCTCG CAGCGCTATTGGC-3' ) (SEQ ID NO:41) sits down on the PRV g50 (gD) 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 EcoRI 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 BcoRI to Sail fragment to generate the approximately 1558 base pair EcoRI to StuI fragment 3. Fragment 4 is an approximately 1484 base pair AccI to Bglll restriction sub-fragment of the SPV Hindlll fragment M (23) .

HOMOLOGY VECTOR 570-91.21. The plasmid 570-91.21 was constructed for the purpose of inserting foreign DNA into SPV. It incorporates an E. coli β-galactosidase (lacZ) marker gene and the PRV gill (gC) gene flanked by SPV DNA. Upstream of the foreign DNA genes is an approximately 1484 base pair fragment of SPV DNA.

Downstream of the foreign genes is an approximately 2149 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 (LPl) , and the gill (gC) gene is under the control of a synthetic early pox promoter (EP2) . A detailed description of the plasmid is given in Figures 10A-10D. 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 Figures 10A-10D. The plasmid vector was derived from an approximately 2972 base pair Hindlll to BamHI restriction fragment of pSP64 (Promega) . Fragment 1 is an approximately 1484 base pair Bglll to AccI restriction sub-fragment of the SPV Hindlll restriction fragment M (23) . Fragment 2 is an approximately 3002 base pair BamHI to PvuII restriction fragment of plasmid pJF751 (11) . Fragment 3 is an approximately 2378 base pair Ncol to Ncol fragment of plasmid 251-41.A, a subfragment of PRV BamHI #2 and #9. BcoRI linkers have replaced the Ncol and Ncol sites at the ends of this fragment. Fragment 4 is an approximately 2149 base pair AccI to Hindlll restriction sub-fragment of the SPV Hindlll fragment M (23) . The AccI sites in fragments 1 and 4 have been converted to PstI sites using synthetic DΝA linkers.

HOMOLOGY VECTOR 570-91.41. The plasmid 570-91.41 was constructed for the purpose of inserting foreign DΝA into SPV. It incorporates an E. coli /3-galactosidase (lacZ) marker gene and the PRV gill (gC) gene flanked by SPV DΝA. Upstream of the foreign DΝA genes is an approximately 2149 base pair fragment of SPV DΝA. Downstream of the foreign genes is an approximately 1484 base pair fragment of SPV DΝA. When this plasmid is used according to the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT SPV, a virus containing DΝA 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 (LPl) , and the gill (gC) gene is under the control of a synthetic early late pox promoter (EP1LP2) . A detailed description of the plasmid is given in Figures 11A-11D. 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 Figures 11A-11D. The plasmid vector was derived from an approximately 2972 base pair Hindlll to BamHI restriction fragment of pSP64 (Promega) . Fragment 1 is an approximately 1484 base pair Bglll to AccI restriction sub-fragment of the SPV Hindlll restriction fragment M (23) . Fragment 2 is an approximately 3002 base pair BamHI to PvuII restriction fragment of plasmid pJF751 (11) . Fragment 3 is an approximately 2378 base pair Ncol to Ncol fragment of plasmid 251-41.A, a subfragment of PRV BamHI #2 and #9. EcoRI linkers have replaced the Ncol and Ncol si tes a t the ends of this fragment . Fragment 4 is an approxima tely 2149 base pair AccI to Hindlll restriction sub-fragment of the SPV Hindlll fragment M

(23) . The AccI si tes in fragments 1 and 4 have been converted to PstI si tes using synthetic DNA linkers .

HOMOLOGY VECTOR 570-91 . 64 . The plasmid 570 -91 . 64 was constructed for the purpose of inserting foreign DNA into SPV. It incorporates an E. coli β-galactosidase (lacZ) marker gene and the PRV gill (gC) gene flanked by SPV DΝA. Upstream of the foreign DΝA genes is an approximately 1484 base pair fragment of SPV DΝA. Downstream of the foreign genes is an approximately 2149 base pair fragment of SPV DΝA. 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 (LPl) , and the gill (gC) gene is under the control of a synthetic late early pox promoter (LP2EP2) . A detailed description of the plasmid is given in Figures 12A-12D. 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 Figures 12A-12D. The plasmid vector was derived from an approximately 2972 base pair Hindlll to BamHI restriction fragment of pSP64 (Promega) . Fragment 1 is an approximately 1484 base pair BglII to AccI restriction sub-fragment of the SPV Hindlll restriction fragment M

(23) . Fragment 2 is an approximately 3002 base pair

BamHI to PvuII restriction fragment of plasmid pJF751 (11) . Fragment 3 is an approximately 2378 base pair Ncol to Ncol fragment of plasmid 251-41.A, a subfragment of

PRV BamHI #2 and #9. EcoRI linkers have replaced the

Νcol and Ncol sites at the ends of this fragment .

Fragment 4 is an approximately 2149 base pair AccI to Hindlll restriction sub-fragment of the SPV Hindlll fragment M (23) . The AccI sites in fragments 1 and 4 have been converted to PstI sites using synthetic DΝA linkers .

HOMOLOGY VECTOR 538-46.26. The plasmid 538-46.26 was constructed for the purpose of inserting foreign DΝA into SPV. It incorporates an E. col i _/S-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 2149 base pair fragment of SPV DNA. Downstream of the foreign genes is an approximately 1484 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 -S-galactosidase (lacZ) marker gene is under the control of a synthetic late pox promoter (LPl) and the HN gene is under the control of a synthetic early/late pox promoter (EP1LP2) . A detailed description of the plasmid is given in Figures 8A-8D. 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 Figures 8A-8D. The plasmid vector was derived from an approximately 2972 base pair Hindlll to BamHI restriction fragment of pSP64

(Promega) . Fragment 1 is an approximately 2149 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 ΝDV HΝ cDΝA clone. The sequence of the HΝ cDΝA clone is given in Figure 7. The cDΝA clone was generated from the Bl strain of ΝDV using standard cDΝA cloning techniques (14) . Fragment 3 is an approximately 3006 base pair BamHI to PvuII restriction fragment of plasmid pJF751 (11) . Fragment 4 is an approximately 1484 base pair AccI to Bglll restriction sub-fragment of the SPV Hindlll fragment M (23) .

HOMOLOGY VECTOR 599-65.25. The plasmid 599-65.25 was constructed for the purpose of inserting foreign DΝA into SPV. It incorporates an E. coli -3-galactosidase (lacZ) marker gene and the ILT gG gene flanked by SPV DΝA. Upstream of the foreign genes is an approximately 1484 base pair fragment of SPV DΝA. Downstream of the foreign genes is an approximately 2149 base pair fragment of SPV DΝA. When the 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 3-galactosidase (lacZ) marker gene is under the control of a synthetic late pox promoter (LPl) , and the ILT gG gene is under the control of a synthetic early/late pox promoter (EP1LP2) . A detailed description of the plasmid is given in Figures 13A-13D. It may be constructed utilizing standard recombinant DNA techniques (22, 30) , by joining restriction fragments from the following sources with the synthetic DNA sequences indicated in Figures 13A-13D. The plasmid vector was derived from an approximately 2972 base pair Hindlll to BamHI restriction fragment of pSP64 (Promega) . Fragment 1 is an approximately 1484 base pair Bglll to AccI restriction sub-fragment of the SPV Hindlll restriction fragment M (23) . Fragment 2 is an approximately 1073 base pair EcoRI to Mbol fragment. Note that the EcoRI site was introduced by PCR cloning. In this procedure, the primers described below were used with a template consisting of a 2.6 kb Sst I to Asp718I subfragment of a 5.1 kb Asp718I fragment of ILT virus genome. The first primer 91.13 (5'- CCGAATTCCGGCTTCAGTAACATAGGATCG -3') (SEQ ID NO: 81) sits down on the ILT gG sequence at amino acid 2. It adds an additional asparagine residue between amino acids 1 and 2 and also introduces an EcoRI restriction site. The second primer 91.14 (5' -GTACCCATACTGGTCGTGGC-3 ' ) (SEQ ID NO: 82) sits down on the opposite strand at approximately amino acid 196 priming toward the 5' end of the gene. The PCR product is digested with EcoRI and BamHI to produce an approximately 454 base pair fragment. The approximately 485 base pair Mbol sub-fragment of ILT Asp718I (5.1 kb) fragment is ligated to the approximately 454 base pair EcoRI to BamHI fragment to generate fragment 2 from EcoRI to Mbol which is approximately 939 base pairs (293 amino acids) in length. Fragment 3 is an approximately 3002 base pair BamHI to PvuII restriction fragment of plasmid pJF751 (11) . Fragment 4 is an approximately 2149 base pair AccI to Hindlll subfragment of the SPV Hindlll fragment M. The AccI sites of fragments 1 and 4 have been converted to PstI sites using synthetic DNA linkers.

HOMOLOGY VECTOR 624-20.IC. The plasmid 624-20.1C was constructed for the purpose of inserting foreign DNA into SPV. It incorporates an E. coli 3-galactosidase (lacZ) marker gene and the ILT gl gene flanked by SPV DNA. Upstream of the foreign genes is an approximately 1484 base pair fragment of SPV DNA. Downstream of the foreign genes is an approximately 2149 base pair fragment of SPV DNA. When the 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 (LPl) , and the ILT gl gene is under the control of a synthetic late/early pox promoter (LP2EP2) . A detailed description of the plasmid is given in Figures 14A-14D. It may be constructed utilizing standard recombinant DNA techniques (22, 30) , by joining restriction fragments from the following sources with the synthetic DNA sequences indicated in Figures 14A-14D. The plasmid vector was derived from an approximately 2972 base pair Hindlll to BamHI restriction fragment of pSP64

(Promega) . Fragment 1 is an approximately 1484 base pair

Bgl II to AccI restriction sub-fragment of the SPV

Hindlll restriction fragment M (23) . Fragment 2 is an approximately 1090 base pair fragment with EcoRI and BamHI restriction sites at the ends synthesized by PCR cloning and containing the entire amino acid coding sequence of the ILT gl gene. The ILT gl gene was synthesized in two separate PCR reactions. In this procedure, the primers described below were used with a template consisting the 8.0 kb ILT Asp 7181 fragment. The first primer 103.6 (5' -CCGGAATTCGCTACTT GGAACTCTGG-3 ' )

(SEQ ID NO: 83) sits down on the ILT gl sequence at amino acid number 2 and introduces an EcoRI site at the 5' end of the ILT gl gene. The second primer 103.3 (5'- CATTGTCCCGAGACGGACAG-3' ) (SEQ ID NO: 84) sits down on the ILT gl sequence at approximately amino acid 269 on the opposite strand to primer 103.6 and primes toward the 5' end of the gene. The PCR product was digested with EcoRI and BglI (BglI is located approximately at amino acid 209 which is 179 base pairs 5' to primer 2) to yield a fragment 625 base pairs in length corresponding to the 5' end of the ILT gl gene. The third primer 103.4 (5'- CGCGATCCAACTATCGGTG-3' ) (SEQ ID NO: 85) sits down on the ILT gl gene at approximately amino acid 153 priming toward the 3' end of the gene. The fourth primer 103.5 (5'GCGGATCCACATTCAG ACTTAATCAC-3' ) (SEQ ID NO: 86) sits down at the 3' end of the ILT gl gene 14 base pairs beyond the UGA stop codon, introducing a BamHI restriction site and priming toward the 5' end of the gene. The PCR product is digested with Bgl I (at amino acid 209) and BamHI to yield a fragment 476 base pairs in length corresponding to the 3' end of the ILT gl gene. Fragment 2 consists of the products of the two PCR reactions ligated together to yield an ILT gl gene which is a EcoRI to BamHI fragment approximately 1101 base pairs (361 amino acids) in length. Fragment 3 is an approximately 3002 base pair BamHI to PvuII restriction fragment of plasmid pJF751 (11) . Fragment 4 is an approximately 2149 base pair AccI to Hindlll subfragment of the SPV Hindlll fragment M. The AccI sites in fragments 1 and 4 were converted to unique NotI sites using NotI linkers.

HOMOLOGY VECTOR 614-83.18. The plasmid 614-83.18 was constructed for the purpose of inserting foreign DΝA into SPV. It incorporates an E. coli _/S-galactosidase (lacZ) marker gene and the IBR gG gene flanked by SPV DΝA.

Upstream of the foreign genes is an approximately 1484 base pair fragment of SPV DΝA. Downstream of the foreign genes is an approximately 2149 base pair fragment of SPV DΝA. When the 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 _/S-galactosidase (lacZ) marker gene is under the control of a synthetic late pox promoter (LPl) , and the IBR gG gene is under the control of a synthetic late/early pox promoter (LP2EP2) . A detailed description of the plasmid is given in Figures 15A-15D. It may be constructed utilizing standard recombinant DNA techniques (22, 30) , by joining restriction fragments from the following sources with the synthetic DNA sequences indicated in Figures 15A-15D. The plasmid vector was derived from an approximately 2972 base pair Hindlll to BamHI restriction fragment of pSP64 (Promega) . Fragment 1 is an approximately 1484 base pair Bgl l l to AccI restriction sub-fragment of the SPV Hindlll restriction fragment M (23) . Fragment 2 is an approximately 1085 base pair fragment synthesized by PCR cloning with EcoRI and BamHI restriction sites at the ends and containing the amino acid coding sequence from amino acids 2 to 362 of the IBR gG gene. In the PCR cloning procedure, the primers described below were used with a template consisting of the IBR-000 virus (Cooper strain) . The first primer 106.9 (5'- ATGAATTCCCCTGCCGCCCGGACCGGCACC-3' ) (SEQ ID NO: 87) sits down on the IBR gG sequence at amino acid number 1 and introduces an EcoRI site at the 5' end of the IBR gG gene and two additional amino acids between amino acids 1 and 2. The second primer 106.8 (5 ' - CATGGATCCCGCTCGAGGCGAGCGGGCTCC-3' ) (SEQ ID NO: 88) sits down on the IBR gG sequence at approximately amino acid 362 on the opposite strand to primer 1 and primes synthesis toward the 5' end of the IBR gG gene. Fragment 2 was generated by digesting the PCR product with EcoRI and BamHI to yield a fragment 1085 base pairs in length corresponding to the amino terminal 362 amino acids

(approximately 80%) of the IBR gG gene. Fragment 3 is an approximately 3002 base pair BamHI to PvuII restriction fragment of plasmid pJF751 (11) . Fragment 4 is an approximately 2149 base pair AccI to Hindlll subfragment of the SPV Hindlll fragment M. The AccI sites in fragments 1 and 4 were converted to unique NotI sites using NotI linkers.

HOMOLOGY VECTOR FOR CONSTRUCTING S-SPV-019 (LacZ/IBR gE HOMOLOGY VECTOR) : This lacZ/IBR gE homology vector is used to insert foreign DNA into SPV. It incorporates an E. coli jβ-galactosidase (lacZ) marker gene and the IBR gE gene flanked by 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 gE gene is under the control of a synthetic late/early pox promoter. The homology vector may be constructed utilizing standard recombinant DNA techniques (22 and 30) , by joining restriction fragments from the following sources with the appropriate synthetic DNA sequences. The plasmid vector is derived from an approximately 2972 base pair Hindlll to BamHI restriction fragment of pSP64 (Promega) . The upstream SPV homology is an approximately 1146 base pair Bgllll to AccI restriction sub-fragment of the SPV Hindlll fragment M (23) . The IBR gE gene is an approximately 1888 base pair fragment synthesized by PCR cloning with EcoRI and BamHI ends. In the PCR cloning procedure, the primers described below were used with a template consisting of the IBR-000 VIRUS (Cooper strain) . The f i rs t p r ime r 4 / 93 . 17 DR ( 5 ' - CTGGTTCGGCCCAGAATTCTATGGGTCTCGCGCGGCTCGTGG-3 ' (SEQ ID NO: 89) sits down on the IBR gE gene at amino acid number 1 and introduces an EcoRI site at the 5' end of the IBR gE gene and adds two additional amino acids at the amino terminus of the protein. The second primer 4/93.18DR (5' - CTCGCTCGCCCAGGATCCCTAGCGGAGGATGGACTTGAGTCG-3' ) (SEQ ID NO: 90) sits down on the IBR gE sequence at approximately amino acid 648 on the opposite strand to primer 1 and primes synthesis toward the 5' end of the IBR gE gene. The lacZ promoter and marker gene is identical to the one used in plasmid 520-17.5. The downstream SPV homology is an approximately 2156 base pair AccI to Hindlll restriction sub-fragment of the SPV Hindlll restriction fragment M (23) . The AccI site in the SPV homology vector is converted to a unique Xbal site.

HOMOLOGY VECTOR FOR CONSTRUCTING S-SPV-018 (LacZ/PRV gE HOMOLOGY VECTOR) : This homology vector is constructed for the purpose of inserting foreign DNA into SPV. It incorporates an E. coli S-galactosidase (lacZ) marker gene and the PRV gE gene flanked by SPV DNA. Upstream of the foreign genes is an approximately 1484 base pair fragment of SPV DNA. Downstream of the foreign genes is an approximately 2149 base pair fragment of SPV DNA. When the plasmid is used according to the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT SPV, a virus containing the DNA coding for the foreign genes results. Note that the /β-galactosidase (lacZ) marker gene is under the control of a synthetic late pox promoter (LPl) , and the PRV gE gene is under the control of a synthetic early/late pox promoter (EP1LP2) . The homology vector is constructed utilizing standard recombinant DNA techniques (22,30), by joining restriction fragments from the following sources with synthetic DNA sequences. The plasmid vector is derived from an approximately 2972 base pair Hindlll to BamHI restriction fragment pSP64 (Promega) . Fragment 1 is an approximately 1484 base pair Bglll to AccI restriction sub-fragment of the SPV Hindlll restriction fragment M

(23) . Fragment 2 is the lacZ promoter and marker gene which is identical to the one used in plasmid 520-17.5.

Fragment 3 is an approximately 2484 base pair Dral to

ΛUuI sub-fragment of PRV derived from the PRV BamHI #7 DNA fragment. The Dral site is converted to an EcoRI site through the use of a synthetic DNA linker. The Dral site sits 45 base pairs upstream of the natural gE start codon and extends the open reading frame at the amino terminus of the protein for 15 amino acids. The synthetic pox promoter/EcoRI DNA linker contributes another 4 amino acids. Therefore, the engineered gE gene contains 19 additional amino acids fused to the amino terminus of gE. The nineteen amino acids are Met-Asn- Ser-Gly-Asn-Leu-Gly-Thr-Pro-Ala-Ser-Leu-Ala-His-Thr-Gly- Val-Glu-Thr. Fragment 4 is an approximately 2149 base pair AccI to Hindlll sub-fragment of the SPV Hindlll fragment M (23) . The AccI sites of fragments 1 and 4 are converted to PstI sites using synthetic DNA linkers.

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 DΝA may be inserted. When a plasmid, containing a foreign DΝA 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) .

HOMOLOGY VECTOR 708-78.9. The plasmid 708-78.9 was constructed for the purpose of inserting foreign DNA into SPV. It incorporates an E. coli _/S-galactosidase (lacZ) marker gene and the infectious bovine rhinotracheitis virus (IBRV) gE gene flanked by SPV DNA. Upstream of the foreign genes is an approximately 1484 base pair fragment of SPV DNA. Downstream of the foreign genes is an approximately 2149 base pair fragment of SPV DNA. When the 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 _/S-galactosidase (lacZ) marker gene is under the control of a synthetic late pox promoter (LPl) , and the IBRV gE gene is under the control of a synthetic late/early pox promoter (LP2EP2) . It may be constructed utilizing standard recombinant DNA techniques (22, 30) , by joining restriction fragments from the following sources. The plasmid vector was derived from an approximately 2972 base pair Hindlll to BamHI restriction fragment of pSP64 (Promega) . Fragment 1 is an approximately 1484 base pair Bgl II to AccI restriction sub-fragment of the SPV Hindlll restriction fragment M (23) . Fragment 2 is an approximately 475 base pair fragment with EcoRI and BamHI restriction sites at the ends. The EcoRI and BamHI sites are synthesized by PCR cloning. The PCR product contains the entire amino acid coding sequence of the IBRV gE gene. In the PCR cloning procedure, the primers described below were used with a template consisting of the IBR-000 virus (Cooper strain)

(44) . The first primer 2/94.5DR (5'-

CTGGTTCGGCCCAGAATTCGATGCAACCCACCGCGCCGCCCCG-3' ) (SEQ ID NO: 116) sits down on the IBR gE gene at amino acid number 1 and introduces an EcoRI site at the 5' end of the IBRV gE gene and adds two additional amino acids at the amino terminus of the protein. The second primer 4/93.18DR (5' -CTCGCTCGCCCAGGATCCCTAGCGGAGGATGGACTTGAGTCG- 3') (SEQ ID NO: 117) sits down on the IBRV gE sequence (44) at approximately amino acid 648 on the opposite strand to the first primer and primes synthesis toward the 5' end of the IBRV gE gene. The PCR product was digested with EcoRI and BamHI to yield a fragment approximately 1950 base pairs in length corresponding to the IBRV gE gene. Fragment 3 is an approximately 3002 base pair BamHI to PvuII restriction fragment of plasmid pJF751 (11) . Fragment 4 is an approximately 2149 base pair AccI to Hindlll subfragment of the SPV Hindlll fragment M. The AccI sites in fragments 1 and 4 were converted to unique NotI sites using NotI linkers.

HOMOLOGY VECTOR 723-59A9.22. The plasmid 723-59A9.22 was used to insert foreign DΝA into SPV. It incorporates an E. coli /S-galactosidase (lacZ) marker gene and the equine influenza virus ΝA PR/56 gene flanked by SPV DΝA. When this plasmid was used according to the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT SPV a virus containing DNA coding for the foreign genes results. Note that the /S-galactosidase (lacZ) marker gene is under the control of a synthetic late pox promoter (LPl) and the EIV PR/56 NA gene is under the control of a synthetic late/early pox promoter (LP2EP2) . A detailed description of the plasmid is given in Figures 18A-18D. The homology vector was constructed utilizing standard recombinant DNA techniques (22 and 30) , by joining restriction fragments from the following sources with the appropriate synthetic DNA sequences. The plasmid vector was derived from an approximately 2972 base pair Hindlll to BamHI restriction fragment of pSP64 (Promega) . Fragment 1 is an approximately 1484 base pair Bglll to AccI restriction sub-fragment of the SPV Hindlll fragment M (23) . Fragment 2 is the NA gene coding region from the equine Influenza A/Prague/56 (serotype 1 (N7) virus) cloned as an approximately 1450 base pair BamHI fragment utilizing the following primers 5 ' - GGGATCCATGAATCCTAATCAAAAACTCTTT-3 ' (SEQ ID NO: 118) for cDNA priming and combined with 5 ' - GGGATCCTTACGAAAAGTATTTAATTTGTGC-3 ' (SEQ ID NO: 119) for PCR. (see CLONING OF EQUINE INFLUENZA VIRUS HEMAGGLUTININ AND NEURAMINIDASE GENES) . Fragment 3 is an approximately 3010 base pair BamHI to PvuII restriction fragment of plasmid pJF751 (11) . Fragment 4 is an approximately 2149 base pair AccI to Hindlll restriction sub-fragment of the SPV Hindlll restriction fragment M (23) . The AccI site in the SPV homology vector was converted to a unique NotI site.

HOMOLOGY VECTOR 727-54.60. The plasmid 727-54.60 was constructed for the purpose of inserting foreign DΝA into SPV. It incorporates an E. col i /S-galactosidase (lacZ) marker gene and the pseudorabies virus (PRV) gll (gB) gene flanked by SPV DΝA. Upstream of the foreign genes is an approximately 1484 base pair fragment of SPV DΝA. Downstream of the foreign genes is an approximately 2149 base pair fragment of SPV DNA. When the 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 (LPl) , and the PRV gB gene is under the control of a synthetic late/early pox promoter (LP2EP2) . A detailed description of the plasmid is given in Figures 19A-19D. It may be constructed utilizing standard recombinant DNA techniques (22, 30) , by joining restriction fragments from the following sources with the synthetic DNA sequences indicated in Figures 19A-19D. The plasmid vector was derived from an approximately 2972 base pair Hindlll to BamHI restriction fragment of pSP64 (Promega) . Fragment 1 is an approximately 1484 base pair Bglll to AccI restriction sub-fragment of the SPV Hindlll restriction fragment M (23) . Fragment 2 is an approximately 3500 base pair fragment which contains the coding sequence for the PRV gB gene within the Kpnl C fragment of genomic PRV DNA(21) . Fragment 2 contains an approximately 53 base pair synthetic fragment containing the amino terminus of the PRV gB gene, an approximately 78 base pair Smal to Nhe I fragment from the PRV Kpnl C genomic fragment, and an approximately 3370 base pair Nhel to EcoRI fragment from the PRV Kpnl C genomic fragment (21) . Fragment 3 is an approximately 3010 base pair BamHI to PvuII restriction fragment of plasmid pJF751 (11) . Fragment 4 is an approximately 2149 base pair AccI to Hindlll subfragment of the SPV Hindlll fragment M. The AccI sites in fragments 1 and 4 were converted to unique NotI sites using Not I linkers.

HOMOLOGY VECTOR 727-67.18. The plasmid 727-67.18 was constructed for the purpose of inserting foreign DΝA into SPV. It incorporates an E. coli _/S-galactosidase (lacZ) marker gene and the hepatitis B virus core antigen gene flanked by SPV DNA. Upstream of the foreign genes is an approximately 1484 base pair fragment of SPV DNA. Downstream of the foreign genes is an approximately 2149 base pair fragment of SPV DNA. When the 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 (LPl) , and the hepatitis B core antigen gene is under the control of a synthetic early/late pox promoter (EP1LP2) . A detailed description of the plasmid is given in Figures 20A-20D. It may be constructed utilizing standard recombinant DNA techniques (22, 30) , by joining restriction fragments from the following sources with the synthetic DNA sequences indicated in Figures 20A-20D. The plasmid vector was derived from an approximately 2972 base pair Hindlll to BamHI restriction fragment of pSP64 (Promega) . Fragment 1 is an approximately 1484 base pair Bglll to AccI restriction sub-fragment of the SPV Hindlll restriction fragment M (23) . Fragment 2 is an approximately 3002 base pair BamHI to PvuII restriction fragment of plasmid pJF751 (11) . Fragment 3 is an approximately 589 base pair fragment with BamHI and EcoRI restriction sites at the ends. This fragment contains the hepatitis B core antigen coding sequences (amino acids 25-212) (Ref. 45, 50) . Fragment 4 is an approximately 2149 base pair AccI to Hindlll subfragment of the SPV Hindlll fragment M. The AccI sites in fragments 1 and 4 were converted to unique Not I sites using Not I linkers.

HOMOLOGY VECTOR 732-18.4. The plasmid 732-18.4 was used to insert foreign DΝA into SPV. It incorporates an E. coli (S-galactosidase (lacZ) marker gene and the equine influenza virus AK/91 ΝA gene flanked by SPV DΝA. When this plasmid was used according to the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT SPV a virus containing DNA coding for the foreign genes results. Note that the /8-galactosidase (lacZ) marker gene is under the control of a synthetic late pox promoter (LPl) and the EIV AK/91 NA gene is under the control of a synthetic late/early pox promoter (LP2EP2) . A detail description of the plasmid is given in Figures 21A-21D. The homology vector was constructed utilizing standard recombinant DNA techniques (22 and 30) , by joining restriction fragments from the following sources with the appropriate synthetic DNA sequences. The plasmid vector was derived from an approximately 2972 base pair Hindlll to BamHI restriction fragment of pSP64 (Promega) . Fragment 1 is an approximately 1484 base pair Bglll to AccI restriction sub-fragment of the SPV Hindlll fragment M (23) . Fragment 2 is the NA gene coding region from the equine Influenza A/Alaska/91 (serotype 2 (N8) virus) cloned as an approximately 1450 base pair Sail fragment utilizing the following primers 5 ' - GGGTCGACATGAATCCAAATCAAAAGATAA-3 ' (SEQ ID NO: 124) for cDNA priming and combined with 5 ' -

GGGTCGACTTACATCTTATCGATGTCAAA-3' (SEQ ID NO: 125) for PCR

(see CLONING OF EQUINE INFLUENZA VIRUS HEMAGGLUTININ AND

NEURAMINIDASE GENES) . Fragment 3 is an approximately 3010 base pair BamHI to PvuII restriction fragment of plasmid pJF751 (11) . Fragment 4 is an approximately 2149 base pair AccI to Hindlll restriction sub-fragment of the SPV Hindlll restriction fragment M (23) . The AccI site in the SPV homology vector was converted to a unique Not I site

HOMOLOGY VECTOR 741-80.3 The plasmid 741-80.3 was constructed for the purpose of inserting foreign DΝA into SPV. It incorporates an E. coli _/S-galactosidase (lacZ) marker gene flanked by SPV DΝA. Upstream of the foreign genes is an approximately 1484 base pair fragment of SPV DΝA. Downstream of the foreign genes is an approximately 2149 base pair fragment of SPV DΝA. When the 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 _/S-galactosidase (lacZ) marker gene is under the control of a human cytomegalovirus immediate early (HCMV IE) promoter. A detailed description of the plasmid is given in Figures 22A-22C. It may be constructed utilizing standard recombinant DNA techniques (22, 30) , by joining restriction fragments from the following sources with the synthetic DNA sequences indicated in Figures 22A-22C.

The plasmid vector was derived from an approximately 2972 base pair Hindlll to BamHI restriction fragment of pSP64

(Promega) . Fragment 1 is an approximately 1484 base pair

Bglll to AccI restriction sub-fragment of the SPV Hindlll restriction fragment M (23) .Fragment 2 is a 1154 base pair PstI to Avail fragment derived from a HCMV 2.1 kb PstI fragment containing the HCMV IE promoter (46) . Fragment 3 is a 3010 base pair BamHI to PvuII fragment derived from plasmid pJF751 (49) containing the E. coli lacZ gene. Fragment 4 is an approximately 750 base pair Ndel to Sail fragment derived from PRV BamHI #7 which contains the carboxy-terminal 19 amino acids and the polyadenylation signal of the PRV gX gene. Fragment 5 is an approximately 2149 base pair AccI to Hindlll subfragment of the SPV Hindlll fragment M. The AccI sites in fragments 1 and 5 were converted to unique Not I sites using NotI linkers.

HOMOLOGY VECTOR 741-84.14. The plasmid 741-84.14 was constructed for the purpose of inserting foreign DΝA into SPV. It incorporates an E. coli β-galactosidase (lacZ) marker gene and the human interleukin-2 (IL-2) gene flanked by SPV DΝA. Upstream of the foreign genes is an approximately 1484 base pair fragment of SPV DΝA. Downstream of the foreign genes is an approximately 2149 base pair fragment of SPV DΝA. When the 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 (LPl) , and the human IL-2 gene is under the control of a synthetic late/early pox promoter (LP2EP2) . The coding sequence for the human IL-2 protein is fused at the amino terminus to the PRV gX signal sequence for membrane transport. A detailed description of the plasmid is given in Figures 23A-23D. It may be constructed utilizing standard recombinant DNA techniques (22, 30) , by joining restriction fragments from the following sources with the synthetic DNA sequences indicated in Figures 23A-23D. The plasmid vector was derived from an approximately 2972 base pair Hindlll to BamHI restriction fragment of pSP64 (Promega) . Fragment 1 is an approximately 1484 base pair Bglll to AccI restriction sub-fragment of the SPV Hindlll restriction fragment M (23) . Fragment 2 is an approximately 475 base pair fragment with EcoRI and Bglll restriction sites at the ends. The EcoRI site is synthesized by PCR cloning and the Bglll site is at the 3' end of the human IL-2 cDNA (43, 47) . The PCR product contains the entire amino acid coding sequence of the PRV gX signal sequence-human IL-2 gene. In this procedure, the primers described below were used with a template consisting of the cDNA for PRV gX signal sequence-human IL-2 (43) . The first primer 5/94.23 (5'- CTCGAATTCGAAGTGGGCAACGTGGATCCTCGC-3' ) (SEQ ID NO: 126) sits down on the PRV gX signal sequence at amino acid number 1 and introduces an EcoRI site at the 5' end of the gene. The second primer 5/94.24 (5'- CAGTTAGCCTCCCCCATCTCCCCA-3' ) (SEQ ID NO: 127) sits down on the human IL-2 gene sequence within the 3' untranslated region on the opposite strand to primer 5/94.23 and primes toward the 5' end of the gene. The PCR product was digested with EcoRI and Bglll (Bglll is located approximately 3 nucleotides beyond the stop codon for the human IL-2 gene) to yield a fragment 475 base pairs in length corresponding to the PRV gX signal sequence-human IL-2 gene. Fragment 3 is an approximately 3010 base pair BamHI to PvuII restriction fragment of plasmid pJF751 (11) . Fragment 4 is an approximately 2149 base pair AccI to Hindlll subfragment of the SPV Hindlll fragment M. The AccI sites in fragments 1 and 4 were converted to unique NotI sites using NotI linkers.

HOMOLOGY VECTOR 744-34. The plasmid 744-34 was constructed for the purpose of inserting foreign DΝA into SPV. It incorporates an E. coli _/S-galactosidase (lacZ) marker gene and the equine herpesvirus type 1 gB gene flanked by SPV DΝA. Upstream of the foreign genes is an approximately 1484 base pair fragment of SPV DΝA. Downstream of the foreign genes is an approximately 2149 base pair fragment of SPV DΝA. When the 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 (LPl) , and the EHV-1 gB gene is under the control of a synthetic late/early pox promoter (LP2EP2) . A detailed description of the plasmid is given in Figures 24A-24D. It may be constructed utilizing standard recombinant DNA techniques (22, 30) , by joining restriction fragments from the following sources with the synthetic DNA sequences indicated in Figures 24A-24D The plasmid vector was derived from an approximately 2972 base pair Hindlll to BamHI restriction fragment of pSP64 (Promega) . Fragment 1 is an approximately 1484 base pair Bgl II to AccI restriction sub-fragment of the SPV Hindlll restriction fragment M (23) . Fragment 2 is an approximately 2941 base pair fragment with EcoRI and Pmel restriction sites at the ends. Fragment 2 is an approximately 2941 base pair EcoRI to Pmel fragment. Fragment 2 was synthesized as an approximately 429 base pair PCR fragment at the 5' end of the gene having a synthetic EcoRI site and a natural BamHI site within the BamHI "a" fragment of EHV-1 genomic DNA and an approximately 2512 base pair restriction fragment at the 3 ' end of the gene from BamHI to Pmel within the BamHI "i" fragment of EHV-1 genomic DNA (48) . In the procedure to produce the 5' end PCR fragment, the primers described below were used with a template consisting of the EHV-1 BamHI "a" and "i" fragments. The first primer 5/94.3 (5' -CGGAATTCCTCTGGTTGCCGT-3 ' ) (SEQ ID NO: 128) sits down on the EHV-1 gB sequence at amino acid number 2 and introduces an EcoRI site at the 5' end of the EHV-1 gB gene and an ATG start codon. The second primer 5/94.4 (5' -GACGGTGGATCCGGTAGGCGGT-3 ' ) (SEQ ID NO: 129) sits down on the EHV-1 gB sequence at approximately amino acid 144 on the opposite strand to primer 5/94.3 and primes toward the 5' end of the gene. The PCR product was digested with EcoRI and BamHI to yield a fragment 429 base pairs in length corresponding to the 5' end of the EHV-1 gB gene. Fragment 2 consists of the products of the PCR reaction (EcoRI to BamHI) and the restriction fragment (BamHI to Pmel) ligated together to yield an EHV-1 gB gene which is an EcoRI to Pmel fragment approximately 2941 base pairs (979 amino acids) in length. Fragment 3 is an approximately 3010 base pair

BamHI to PvuII restriction fragment of plasmid pJF751

(11) . Fragment 4 is an approximately 2149 base pair AccI to Hindlll subfragment of the SPV Hindlll fragment M. The

AccI sites in fragments 1 and 4 were converted to unique NotI sites using NotI linkers.

HOMOLOGY VECTOR 744-38. The plasmid 744-38 was constructed for the purpose of inserting foreign DΝA into SPV. It incorporates an E. coli _/8-galactosidase (lacZ) marker gene and the equine herpesvirus type 1 gD gene flanked by SPV DΝA. Upstream of the foreign genes is an approximately 1484 base pair fragment of SPV DΝA. Downstream of the foreign genes is an approximately 2149 base pair fragment of SPV DNA. When the 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 (LPl) , and the EHV-1 gD gene is under the control of a synthetic late/early pox promoter (LP2EP2) . A detailed description of the plasmid is given in Figures 25A-25D. It may be constructed utilizing standard recombinant DNA techniques (22, 30) , by joining restriction fragments from the following sources with the synthetic DNA sequences indicated in Figures 25A-25D. The plasmid vector was derived from an approximately 2972 base pair Hindlll to BamHI restriction fragment of pSP64 (Promega) . Fragment 1 is an approximately 1484 base pair Bgl II to AccI restriction sub-fragment of the SPV Hindlll restriction fragment M (23) . Fragment 2 is an approximately 1240 base pair Hindlll fragment within the BamHI "d" fragment of EHV-1 (48) . Fragment 3 is an approximately 3010 base pair BamHI to PvuII restriction fragment of plasmid pJF751 (11) . Fragment 4 is an approximately 2149 base pair AccI to Hindlll subfragment of the SPV Hindlll fragment M. The AccI sites in fragments 1 and 4 were converted to unique

NotI sites using NotI linkers.

HOMOLOGY VECTOR 689-50.4. The plasmid 689-50.4 was constructed for the purpose of inserting foreign DΝA into SPV. It incorporates an E. coli _/S-galactosidase (lacZ) marker gene and the infectious bursal disease virus (IBDV) polyprotein gene flanked by SPV DΝA. Upstream of the foreign genes is an approximately 1484 base pair fragment of SPV DΝA. Downstream of the foreign genes is an approximately 2149 base pair fragment of SPV DΝA.

When the 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 S-galactosidase (lacZ) marker gene is under the control of a synthetic late pox promoter (LPl) , and the IBDV polyprotein gene is under the control of a synthetic late/early pox promoter (LP2EP2) . It may be constructed utilizing standard recombinant DNA techniques (22, 30), by joining restriction fragments from the following sources. The plasmid vector was derived from an approximately 2972 base pair Hind III to BamHI restriction fragment of pSP64 (Promega) . Fragment 1 is an approximately 1484 base pair Bglll to AccI restriction subfragment of the SPV Hindlll restriction fragment M (23) . Fragment 2 is an approximately 3400 base pair fragment with S al and Hpal restriction sites at the ends from plasmid 2-84/2-40 (51) . This fragment contains the IBDV polyprotein coding sequences. Fragment 3 is an approximately 3010 base pair BamHI to PvuII restriction fragment of plasmid pJF751 (11) . Fragment 4 is an approximately 2149 base pair AccI to Hindlll subfragment of the SPV Hindlll fragment M. The AccI sites in fragments 1 and 4 were converted to unique NotI sites using NotI linkers.

HOMOLOGY VECTOR 689-50.7. The plasmid 689-50.7 was constructed for the purpose of inserting foreign DΝA into

SPV. It incorporates an E. coli _/S-galactosidase (lacZ) marker gene and the infectious bursal disease virus

(IBDV) VP2 gene flanked by SPV DΝA. Upstream of the foreign genes is an approximately 1484 base pair fragment of SPV DΝA. Downstream of the foreign genes is an approximately 2149 base pair fragment of SPV DΝA. When the 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 _/S-galactosidase (lacZ) marker gene is under the control of a synthetic late pox promoter

(LPl) , and the IBDV VP2 gene is under the control of a synthetic late/early pox promoter (LP2EP2) . It may be constructed utilizing standard recombinant DNA techniques (22, 30) , by joining restriction fragments from the following sources. The plasmid vector was derived from an approximately 2972 base pair Hindlll to BamHI restriction fragment of pSP64 (Promega) . Fragment 1 is an approximately 1484 base pair Bglll to AccI restriction sub-fragment of the SPV Hindlll restriction fragment M (23) . Fragment 2 is an approximately 1081 base pair fragment with Bell and BamHI restriction sites at the ends. This fragment codes for the IBDV VP2 protein and is derived from a full length IBDV cDNA clone (51) . Fragment 3 is an approximately 3010 base pair BamHI to PvuII restriction fragment of plasmid pJF751 (11) . Fragment 4 is an approximately 2149 base pair AccI to Hindlll sub-fragment of the SPV Hindlll fragment M. The AccI sites in fragments 1 and 4 were converted to unique NotI sites using NotI linkers.

HOMOLOGY VECTOR 751-07.Al. The plasmid 751-07.Al was used to insert foreign DΝA into SPV. It incorporates an E. coli S-galactosidase (lacZ) marker gene and the chicken interferon (cIFΝ) gene flanked by SPV DΝA. When this plasmid was used according to the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT SPV a virus containing DNA coding for the foreign genes results. Note that the _/S-galactosidase (lacZ) marker gene is under the control of a synthetic late pox promoter

(LPl) and the cIFN gene is under the control of a synthetic late/early pox promoter (LP2EP2) . The homology vector was constructed utilizing standard recombinant DNA techniques (22 and 30) , by joining restriction fragments from the following sources with the appropriate synthetic DNA sequences. The plasmid vector was derived from an approximately 2972 base pair Hindlll to BamHI restriction fragment of pSP64 (Promega) . Fragment 1 is an approximately 1146 base pair Bglll to AccI restriction sub-fragment of the SPV Hindlll fragment M (23) . Fragment 2 is an approximately 577 base pair EcoRI to Bglll fragment coding for the cIFN gene (54) derived by reverse transcription and polymerase chain reaction (PCR) (Sambrook, et al. , 1989) of RNA ISOLATED FROM CONCANAVALIN A STIMULATED CHICKEN SPLEEN CELLS. The antisense primer (6/94.13) used for reverse transcription and PCR was 5' -CGACGGATCCGAGGTGCGTTTGGGGCTAAGTGC-3 ' (SEQ ID NO: 211) . The sense primer (6/94.12) used for PCR was 5' -CCACGGATCCAGCACAACGCGAGTCCCACCATGGCT-3' (SEQ ID NO: 212) . The BamHI fragment resulting from reverse transcription and PCR was gel purified and used as a template for a second PCR reaction to introduce a unique EcoRI site at the 5' end and a unique Bglll site at the 3' end. The second PCR reaction used primer 6/94.22 (5'- CCACGAATTCGATGGCTGTGCCTGCAAGCCCACAG-3' ; SEQ ID NO: 213) at the 5' end and primer 6/94.34 (5'- CGAAGATCTGAGGTGCGTTTGGGGCTAAGTGC-3' ; SEQ ID NO: 214) at the 3' end to yield an approximately 577 base pair fragment. The DNA fragment contains the coding sequence from amino acid 1 to amino acid 193 of the chicken interferon protein (54) which includes a 31 amino acid signal sequence at the amino terminus and 162 amino acids of the mature protein encoding chicken interferon. Fragment 3 is an approximately 3002 base pair BamHI to PvuII restriction fragment of plasmid pJF751 (11) . Fragment 4 is an approximately 2156 base pair AccI to Hindlll restriction sub-fragment of the SPV Hindlll restriction fragment M (23) . The AccI site in the SPV homology vector was converted to a unique NotI site.

HOMOLOGY VECTOR 751-56.Al. The plasmid 751-56.Al was used to insert foreign DNA into SPV. It incorporates an E. coli _/S-galactosidase (lacZ) marker gene and the chicken myelomonocytic growth factor (cMGF) gene flanked by SPV DNA. When this plasmid was used according to the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT SPV a virus containing DΝA coding for the foreign genes results. Note that the _/S-galactosidase (lacZ) marker gene is under the control of a synthetic late pox promoter (LPl) and the cMGF gene is under the control of a synthetic late/early pox promoter (LP2EP2) . The homology vector was constructed utilizing standard recombinant DNA techniques (22 and 30) , by joining restriction fragments from the following sources with the appropriate synthetic DNA sequences. The plasmid vector was derived from an approximately 2972 base pair Hindlll to BamHI restriction fragment of pSP64 (Promega) . Fragment 1 is an approximately 1146 base pair Bglll to AccI restriction sub-fragment of the SPV Hindlll fragment M (23) . Fragment 2 is an approximately 640 base pair EcoRI to BamHI fragment coding for the cMGF gene (55) derived by reverse transcription and polymerase chain reaction (PCR) (Sambrook, et al . , 1989) of RNA ISOLATED FROM CONCANAVALIN A STIMULATED CHICKEN SPLEEN CELLS. The antisense primer (6/94.20) used for reverse transcription and PCR was 5' -CGCAGGATCCGGGGCGTCAGAGGCGGGCGAGGTG-3 '

(SEQ ID NO: 215) . The sense primer (5/94.5) used for PCR was 5' -GAGCGGATCCTGCAGGAGGAGACACAGAGCTG-3' (SEQ ID NO: 216) . The BamHI fragment derived from PCR was subcloned into a plasmid and used as a template for a second PCR reaction using primer 6 /94.16 ( 5 ' - GCGCGAATTCCATGTGCTGCCTCACCCCTGTG-3 ' ; SEQ ID NO: 217) at the 5' end and primer 6/94.20 (5' - CGCAGGATCCGGGGCGTCAGAGGCGGGCGAGGTG-3 ' ; SEQ ID NO: 218) at the 3' end to yield an approximately 640 base pair fragment. The DNA fragment contains the coding sequence from amino acid 1 to amino acid 201 of the cMGF protein (55) which includes a 23 amino acid signal sequence at the amino terminus and 178 amino acids of the mature protein encoding cMGF. Fragment 3 is an approximately 3002 base pair BamHI to PvuII restriction fragment of plasmid pJF751 (11) . Fragment 4 is an approximately 2156 base pair AccI to Hindlll restriction sub-fragment of the SPV Hindlll restriction fragment M (23) . The AccI site in the SPV homology vector was converted to a unique NotI site.

HOMOLOGY VECTOR 752-22.1. The plasmid 752-22.1 was constructed for the purpose of inserting foreign DNA into SPV. It incorporates an E. coli S-galactosidase (lacZ) marker gene flanked by SPV DNA. Upstream of the foreign gene is an approximately 855 base pair fragment of SPV DNA. Downstream of the foreign genes is an approximately 1113 base pair fragment of SPV DNA. When the 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 _/S-galactosidase (lacZ) marker gene is under the control of a swinepox virus OIL gene promoter. The homology vector also contains the synthetic late/early promoter

(LP2EP2) into which a second foreign gene is inserted into a unique BamHI or EcoRI site. A detailed description of the plasmid is given in Figures 28A-28D. It was constructed utilizing standard recombinant DNA techniques (22, 30) , by joining restriction fragments from the following sources with the synthetic DNA sequences indicated in Figures 28A-28D. The plasmid vector was derived from an approximately 2519 base pair Hindlll to SphI restriction fragment of pSP65 (Promega) . Fragment 1 is an approximately 855 base pair sub-fragment of the SPV Hindlll restriction fragment M (23) synthesized by polymerase chain reaction using DNA primers 5'- GAAGCATGCCCGTTCTTATCAATAGTTTAGTCGAAAATA-3' (SEQ ID NO:

185) and 5' -CATAAGATCTGGCATTGTGTTATTATACTAACAAAAATAAG-3 '

(SEQ ID NO: 186) to produce an 855 base pair fragment with SphI and Bglll ends. Fragment 2 is a 3002 base pair

BamHI to PvuII fragment derived from plasmid pJF751 (49) containing the E. coli lacZ gene. Fragment 3 is an approximately 1113 base pair subfragment of the SPV Hindlll fragment M synthesized by polymerase chain r e a c t i o n u s i ng D NA p r i m e r s 5 ' - CCGTAGTCGACAAAGATCGACTTATTAATATGTATGGGATT-3 ' (SEQ ID NO: 187) and 5' -GCCTGAAGCTTCTAGTACAGTATTTACGACTTTTGAAAT-3' (SEQ ID NO: 188) to produce an 1113 base pair fragment with Sail and Hindlll ends.

HOMOLOGY VECTOR 752-29.33. The plasmid 759.33 was constructed for the purpose of inserting foreign DNA into SPV. It incorporates an E. coli _/S-galactosidase (lac Z) marker gene and an equine herpesvirus type 1 gB gene flanked by SPV DNA. Upstream of the foreign gene is an approximately 855 base pair fragment of SPV DNA. Downstream of the foreign genes is an approximately 113 base pair fragment of SPV DNA. When the 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 swinepox virus OIL gene promoter and the EHV-1 gB gene is under the control of the late/early promoter (LP2EP2) . The LP2EP2 promoter-EHV-1 gB gene cassette was inserted into a NotI site of homology vector 738-94.4. Homology vector 752-29.33 was constructed utilizing standard recombinant DNA techniques (22, 30) , by joining restriction fragments from the following sources with the synthetic DNA sequences. The plasmid vector was derived from an approximately 2519 base pair Hindlll to SphI restriction fragment of pSP65 (Promega) . Fragment 1 is an approximately 855 base pair sub-fragment of the SPV Hindlll restriction fragment M (23) synthesized by polymerase chain reaction using DNA primers 5'-

GAAGCATGCCCGTTCTTATCAATAGTTTAGTCGAAAATA-3' (SEQ ID NO:

185) and 5' -CATAAGATCTGGCATTGTGTTATTATACTAACAAAAATAAG-3 '

(SEQ ID NO: 186) to produce an 855 base pair fragment with SphI and Bglll ends. Fragment 2 is a 3002 base pair BamHI to PvuII fragment derived from plasmid pJF751 (49) containing the E. coli lacz gene. Fragment 3 is the product of a PCR reaction (EcoRI to BamHI) and a restriction fragment (BamHI to Pmel) ligated together to yield an EHV-1 gB gene which is an EcoRI to Pmel fragment approximately 2941 base pairs (979 amino acids) in length. The PCR fragment is an approximately 429 base pair fragment having a synthetic EcoRI site at the 5' end of the gene and a natural BamHI site at the 3 ' end within the BamHI "a" fragment of EHV-1 genomic DNA. The restriction fragment is an approximately 2512 base pair fragment from BamHI to Pmel within the BamHI "I" fragment of EHV-1 genomic DNA. In the procedure to produce the 5' end PCR fragment, the primers described below were used with a template consisting of the EHV-1 BamHI "a" and "i" fragments.

The first primer 5/94.3 (5' -CGGAATTCCTCTGGTTCGCCGT-3 ' ) (SEQ ID NO: 128) sits down on the EHV-1 gB sequence at amino acid number 2 and introduces an EcoRI site at the 5' end of the EHV-1 gB gene and an ATG start codon. The second primer 5/94.4 (5' -GACGGTGGATCCGGTAGGCGGT-3 ' ) (SEQ ID NO: 129) sits down on the EHV-1 gB sequence at approximately amino acid 144 on the opposite strand to primer 5/94.3 and primes toward the 5' end of the gene. The PCR product was digested with EcoRI and BamHI to yield a fragment 429 base pairs in length corresponding to the 5' end of the EHV-1 gB gene. Fragment 3 consists of the products of the PCR reaction (EcoRI to BamHI) and the restriction fragment (BamHI to Pmel) ligated together to yield an EHV-1 gB gene which is an EcoRI to Pmel fragment approximately 2941 base pairs (979 amino acids) in length. Fragment 4 is an approximately 1113 base pair subfragment of the SPV Hindlll fragment M synthesized by polymerase chain reaction using DNA primers 5'- CCGTAGTCGACAAAGATCGACTTATTAATATGTATGGGATT-3' (SEQ ID NO: 187) and 5' -GCCTGAAGCTTCTAGTACAGTATTTACGACTTTTGAAAT-3 ' (SEQ ID NO: 188) to produce an 1113 base pair fragment with Sail and Hindlll ends. HOMOLOGY VECTOR 746-94.1. The plasmid 746-94.1 was constructed for the purpose of inserting foreign DNA into SPV. It incorporates an E. coli S-galactosidase (lacZ) marker gene and an infectious bovine rhinotracheitis virus glycoprotein E (gE) gene flanked by SPV DNA. Upstream of the foreign gene is an approximately 855 base pair fragment of SPV DNA. Downstream of the foreign genes is an approximately 1113 base pair fragment of SPV DNA. When the 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 _/S-galactosidase (lacZ) marker gene is under the control of a swinepox virus OIL gene promoter and the IBRV gE gene is under the control of the late/early promoter (LP2EP2) . It was constructed utilizing standard recombinant DNA techniques (22, 30) , by joining restriction fragments from the following sources with the synthetic DNA sequences. A 1250 base pair EcoRI to BamHI fragment coding for amino acids 1 to 417 of the IBRV gE gene (missing 158 amino acids of the carboxy terminal transmembrane region) was inserted into unique EcoRI and BamHI sites of homology vector 752-22.1

(Figures 28A-28D) . The 1250 base pair EcoRI to BamHI fragment was synthesized by polymerase chain reaction (15) using IBRV (Cooper) genomic DNA as a template and primer 10/94.23 (5' -GGGGAATTCAATGCAACCCACCGCGCCGCCCC-3 ' ; SEQ ID NO: 219) at the 5' end of the IBRV gE gene (amino acid 1 ) and primer 10/94.22 ( 5 ' - GGGGGATCCTAGGGCGCGCCCGCCGGCTCGCT-3' ; SEQ ID NO: 220) at amino acid 417 of the IBRV gE gene.

HOMOLOGY VECTOR 767-67.3. The plasmid 767-67.3 was constructed for the purpose of inserting foreign DNA into SPV. It incorporates an E. coli _/S-galactosidase (lacZ) marker gene and an bovine viral diarrhea virus glycoprotein 53 (BVDV gp53) gene flanked by SPV DNA. Upstream of the foreign gene is an approximately 855 base pair fragment of SPV DNA. Downstream of the foreign genes is an approximately 1113 base pair fragment of SPV DNA. When the 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 /S-galactosidase (lacZ) marker gene is under the control of a swinepox virus OIL gene promoter and the BVDV gp53 gene is under the control of the late/early promoter (LP2EP2) . It was constructed utilizing standard recombinant DNA techniques (22, 30) , by joining restriction fragments from the following sources with the synthetic DNA sequences. A 1187 base pair BamHI fragment coding for the BVDV gp53 was inserted into the unique BamHI sites of homology vector 752-22.1 (Figures 28A-28D) . The 1187 base pair BamHI fragment was synthesized by polymerase chain reaction (15) as described in CLONING OF BOVINE VIRAL DIARRHEA VIRUS gp48 AND gp53 GENES.

HOMOLOGY VECTOR 771-55.11. The plasmid 771-55.11 was constructed for the purpose of inserting foreign DNA into SPV. It incorporates an E. coli _/β-galactosidase (lacZ) marker gene and an bovine viral diarrhea virus glycoprotein 48 (BVDV gp48) gene flanked by SPV DNA. Upstream of the foreign gene is an approximately 855 base pair fragment of SPV DNA. Downstream of the foreign genes is an approximately 1113 base pair fragment of SPV DNA. When the 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 /S-galactosidase (lacZ) marker gene is under the control of a swinepox virus OIL gene promoter and the BVDV gp48 gene is under the control of the late/early promoter (LP2EP2) . It was constructed utilizing standard recombinant DNA techniques (22, 30) , by joining restriction fragments from the following sources with the synthetic DNA sequences. A 678 base pair BamHI fragment coding for the BVDV gp48 was inserted into the unique BamHI sites of homology vector 752-22.1 (Figures 28A-28D) . The 678 base pair BamHI fragment was synthesized by polymerase chain reaction (15) as described in CLONING OF BOVINE VIRAL DIARRHEA VIRUS gp48 AND gp53 GENES.

PLASMID 551-47.23. The plasmid 551-47.23 was constructed for the purpose of inserting foreign DNA into SPV. It incorporates the E. coli _/S-glucuronidase (/S-glu) marker gene under the control of a late/early pox promoter

(LP2EP2) . It is useful to insert the marker gene into sites in the SPV genome to produce a recombinant swinepox virus. It was constructed utilizing standard recombinant DNA techniques (22, 30) , by joining restriction fragments from the following sources. The plasmid vector was derived from an approximately 3005 base pair Hindlll restriction fragment of pSP65 (Promega) . Fragment 1 is an approximately 1823 base pair EcoRI to Smal fragment of the plasmid pRAJ260 (Clonetech) . Note that the EcoRI and Smal sites were introduced by PCR cloning. Plasmid 551- 47.23 was used to make recombinant swinepox viruses S- SPV-059, S-SPV-060, S-SPV-061, and S-SPV-062.

HOMOLOGY VECTOR 779-94.31. The plasmid 779-94.31 was constructed for the purpose of inserting foreign DNA into SPV. It incorporates an E. coli _/3-galactosidase (lacZ) marker gene and the pseudorabies virus (PRV) gB (gll) gene flanked by SPV DNA. Upstream of the foreign genes is an approximately 538 base pair fragment of SPV DNA. Downstream of the foreign genes is an approximately 1180 base pair fragment of SPV DNA. When the 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 (LPl) and the PRV gB gene is under the control of a synthetic late/early pox promoter (LP2EP2) . A detailed description of the plasmid is given in Figures 30A-30E. It was constructed utilizing standard recombinant DNA techniques (22, and 30) , by joining restriction fragments from the following sources with the synthetic DNA sequences. The plasmid vector was derived from an approximately 2986 base pair Hindlll to PstI restriction fragment of pSP64 (Promega) . Fragment 1 is an approximately 542 base pair Hindlll to Bglll restriction sub-fragment of the SPV Hindlll restriction fragment M (23) . Fragment 2 is an approximately 3500 base pair fragment which contains the coding sequence for the

PRV gB gene within the Kpnl C fragment of genomic PRV DNA

(21) . Fragment 2 contains an approximately 53 base pair synthetic fragment containing the amino terminus of the PRV gB gene, an approximately 78 base pair Smal to Nhe I fragment from the PRV Kpnl C genomic fragment, and an approximately 3370 base pair Nhel to EcoRI fragment from the PRV Kpnl C genomic fragment (21) . Fragment 3 is an approximately 3010 base pair BamHI to PvuII restriction fragment of plasmid pJF751 (11) . Fragment 4 is an approximately 1180 base pair Bglll to PstI subfragment of the SPV Hindlll fragment M. The Bglll sites in fragments 1 and 4 were converted to unique Hindlll sites using Hindlll linkers.

HOMOLOGY VECTOR 789-41.7. The plasmid 789-41.7 was constructed for the purpose of inserting foreign DNA into SPV. It incorporates an E. coli /8-galactosidase (lacZ) marker gene, the pseudorabies virus (PRV) gB (gll) gene and the PRV gD (g50) gene flanked by SPV DNA. Upstream of the foreign genes is an approximately 1484 base pair fragment of SPV DNA. Downstream of the foreign genes is an approximately 1560 base pair fragment of SPV DNA. When the 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 (LPl) , the PRV gB gene is under the control of a synthetic late/early pox promoter (LP2EP2) , and the PRV gD gene is under the control of a synthetic early/late pox promoter (EP1LP2) . A detailed description of the plasmid is given in Figures 31A-31D. It was constructed utilizing standard recombinant DNA techniques (22, 30) , by joining restriction fragments from the following sources with the synthetic DNA sequences. The plasmid vector was derived from an approximately 2972 base pair Hindlll to BamHI restriction fragment of pSP64 (Promega) . Fragment 1 is an approximately 1484 base pair Bglll to AccI restriction sub-fragment of the SPV Hindlll restriction fragment M (23) . Fragment 2 is an approximately 1552 base pair subfragment of the PRV BamHI #7 fragment which contains the coding sequence of the PRV gD gene from amino acid 3 to amino acid 279. The EcoRI site and the ATG translation start codon are derived from a polymerase chain reaction using a 5' primer with an EcoRI site. The StuI site at the 3' end is normally within the PRV gl gene 3' to the PRV gD gene. The entire open reading frame beginning at the EcoRI site codes for 405 amino acids. Fragment 3 is an approximately 48 base pair AccI to Ndel subfragment of the SPV Hindlll M fragment. Fragment 4 is an approximately 3500 base pair fragment which contains the coding sequence for the PRV gB gene within the Kpnl C fragment of genomic PRV DNA(21) . Fragment 4 contains an approximately 53 base pair synthetic fragment containing the amino terminus of the PRV gB gene, an approximately 78 base pair Smal to Nhe I fragment from the PRV Kpnl C genomic fragment, and an approximately 3370 base pair Nhel to EcoRI fragment from the PRV Kpnl C genomic fragment (21) . Fragment 5 is an approximately 3010 base pair BamHI to PvuII restriction fragment of plasmid pJF751 (11) . Fragment 6 is an approximately 1560 base pair Ndel to Hindlll subfragment of the SPV Hindlll fragment M. The AccI sites in fragments 1 and 3 were converted to unique PstI sites using PstI linkers. The Ndel sites in fragments 3 and 6 were converted to unique Hindlll sites using Hindlll linkers. An approximately 545 base pair Ndel to Ndel subfragment (Nucleotides 1560 to 2104; SEQ ID NO. ) of the SPV Hindlll M fragment has been deleted which would span SPV fragments 3 and 6.

HOMOLOGY VECTOR 789-41.27. The plasmid 789-41.27 was constructed for the purpose of inserting foreign DNA into SPV. It incorporates an E. coli _/S-galactosidase (lacZ) marker gene, the pseudorabies virus (PRV) gB (gll) gene and the PRV gC (gill) gene flanked by SPV DNA. Upstream of the foreign genes is an approximately 1560 base pair fragment of SPV DNA. Downstream of the foreign genes is an approximately 1484 base pair fragment of SPV DNA. When the 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 (LPl) , the PRV gB gene is under the control of a synthetic late/early pox promoter (LP2EP2) , and the PRV gC gene is under the control of a synthetic early/late pox promoter (EP1LP2) . A detailed description of the plasmid is given in Figures 32A-32D. It was constructed utilizing standard recombinant DNA techniques (22, 30) , by joining restriction fragments from the following sources with the synthetic DNA sequences indicated. The plasmid vector was derived from an approximately 2972 base pair Hindlll to BamHI restriction fragment of pSP64

(Promega) . Fragment 1 is an approximately 1560 base pair

Hindlll to Ndel subfragment of the SPV Hindlll fragment M. Fragment 2 is an approximately 3500 base pair fragment which contains the coding sequence for the PRV gB gene within the Kpnl C fragment of genomic PRV DNA(21) . Fragment 2 contains an approximately 53 base pair synthetic fragment containing the amino terminus of the PRV gB gene, an approximately 78 base pair Smal to Nhe I fragment from the PRV Kpnl C genomic fragment, and an approximately 3370 base pair Nhel to EcoRI fragment from the PRV Kpnl C genomic fragment (21) . Fragment 3 is an approximately 3010 base pair BamHI to PvuII restriction fragment of plasmid pJF751 (11) . Fragment 4 is an approximately 48 base pair AccI to Ndel subfragment of the SPV Hindlll M fragment. Fragment 5 is an approximately 2378 base pair Ncol to Ncol fragment of plasmid 251-41.A, a subfragment of PRV BamHI #2 and #9. EcoRI linkers have replaced the Ncol sites at the ends of the fragment. Fragment 6 is an approximately 1484 base pair AccI to Bglll restriction sub-fragment of the SPV Hindlll restriction fragment M (23) . The Ndel sites in fragments 1 and 4 were converted to unique Hindlll sites using Hindlll linkers. The AccI site in fragments 4 and 6 were converted to unique PstI sites using PstI linkers. An approximately 545 base pair Ndel to Ndel (Nucleotides 1560 to 2104; SEQ ID NO. ) subfragment of the SPV Hindlll M fragment has been deleted which would span SPV fragments 4 and 6.

HOMOLOGY VECTOR 789-41.47. The plasmid 789-41.47 was constructed for the purpose of inserting foreign DNA into SPV. It incorporates an E. coli 0-galactosidase (lacZ) marker gene, the pseudorabies virus (PRV) gC (gill) gene and the PRV gD (g50) gene flanked by SPV DNA. Upstream of the foreign genes is an approximately 1484 base pair fragment of SPV DNA. Downstream of the foreign genes is an approximately 1560 base pair fragment of SPV DNA. When the 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 (LP1) , the PRV gC gene is under the control of a synthetic early/late pox promoter (EP1LP2) , and the PRV gD gene is under the control of a synthetic early/late pox promoter (EP1LP2) . A detailed description of the plasmid is given in Figures 33A-33D. It was constructed utilizing standard recombinant DNA techniques (22, 30) , by joining restriction fragments from the following sources with the synthetic DNA sequences. The plasmid vector was derived from an approximately 2972 base pair Hindlll to BamHI restriction fragment of pSP64 (Promega) . Fragment 1 is an approximately 1484 base pair Bglll to AccI restriction sub-fragment of the SPV Hindlll restriction fragment M (23) . Fragment 2 is an approximately 1552 base pair subfragment of the PRV BamHI #7 fragment which contains the coding sequence of the PRV gD gene from amino acid 3 to amino acid 279. The EcoRI site and the ATG translation start codon are derived from a polymerase chain reaction using a 5' primer with an EcoRI site. The StuI site at the 3' end is normally within the PRV gl gene 3' to the PRV gD gene. The entire open reading frame beginning at the EcoRI site codes for 405 amino acids. Fragment 3 is an approximately 48 base pair AccI to Ndel subfragment of the SPV Hindlll M fragment. Fragment 4 is an approximately 3010 base pair BamHI to PvuII restriction fragment of plasmid pJF751 (11) . Fragment 5 is an approximately 2378 base pair Ncol to Ncol fragment of plasmid 251-41.A, a subfragment of PRV BamHI #2 and #9. EcoRI linkers have replaced the Ncol sites at the ends of the fragment. Fragment 6 is an approximately 1560 base pair Ndel to Hindlll subfragment of the SPV Hindlll fragment M. The AccI sites in fragments 1 and 3 were converted to unique PstI sites using PstI linkers. The Ndel sites in fragments 3 and 6 were converted to unique Hindlll sites using Hindlll linkers. An approximately 545 base pair Ndel to Ndel subfragment (Nucleotides 1560 to 2104; SEQ ID NO. ) of the SPV Hindlll M fragment has been deleted which would span SPV fragments 3 and 6.

HOMOLOGY VECTOR 789-41.73. The plasmid 789-41.73 was constructed for the purpose of inserting foreign DNA into SPV. It incorporates an E. coli _/β-galactosidase (lacZ) marker gene, the pseudorabies virus (PRV) gB (gll) gene, the PRV gC (gill) gene and the PRV gD (g50) gene flanked by SPV DNA. Upstream of the foreign genes is an approximately 1484 base pair fragment of SPV DNA. Downstream of the foreign genes is an approximately 1560 base pair fragment of SPV DNA. When the 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 (LPl) , the PRV gB gene is under the control of a synthetic late/early pox promoter

(LP2EP2) , the PRV gC gene is under the control of a synthetic early/late promoter (EP1LP2) , and the PRV gD gene is under the control of a synthetic late/early pox promoter (LP2EP2) . A detailed description of the plasmid is given in Figures 34A-34E. It was constructed utilizing standard recombinant DNA techniques (22, 30) , by joining restriction fragments from the following sources with the synthetic DNA sequences. The plasmid vector was derived from an approximately 2972 base pair Hindlll to BamHI restriction fragment of pSP64 (Promega) . Fragment 1 is an approximately 1484 base pair Bglll to AccI restriction sub-fragment of the SPV Hindlll restriction fragment M (23) . Fragment 2 is an approximately 1552 base pair subfragment of the PRV BamHI #7 fragment which contains the coding sequence of the PRV gD gene from amino acid 3 to amino acid 279. The EcoRI site and the ATG translation start codon are derived from a polymerase chain reaction using a 5' primer with an EcoRI site. The StuI site at the 3' end is normally within the PRV gl gene 3' to the PRV gD gene. The entire open reading frame beginning at the EcoRI site codes for 405 amino acids. Fragment 3 is an approximately 2378 base pair Ncol to Ncol fragment of plasmid 251-41.A, a subfragment of PRV BamHI #2 and #9. EcoRI linkers have replaced the Ncol sites at the ends of the fragment. Fragment 4 is an approximately 48 base pair AccI to Ndel subfragment of the SPV Hindlll M fragment. Fragment 5 is an approximately 3500 base pair fragment which contains the coding sequence for the PRV gB gene within the Kpnl C fragment of genomic PRV DNA(21) . Fragment 5 contains an approximately 53 base pair synthetic fragment containing the amino terminus of the PRV gB gene, an approximately 78 base pair Smal to Nhe I fragment from the PRV Kpnl C genomic fragment, and an approximately 3370 base pair Nhel to EcoRI fragment from the PRV Kpnl C genomic fragment (21) . Fragment 6 is an approximately 3010 base pair BamHI to PvuII restriction fragment of plasmid pJF751 (11) . Fragment 7 is an approximately 1560 base pair Ndel to Hindlll subfragment of the SPV Hindlll fragment M. The AccI sites in fragments 1 and 3 were converted to unique PstI sites using PstI linkers. The Ndel sites in fragments 3 and 6 were converted to unique Hindlll sites using Hindlll linkers. An approximately 545 base pair Ndel to Ndel subfragment (Nucleotides 1560 to 2104; SEQ ID NO. ) of the SPV Hindlll M fragment has been deleted which would span SPV fragments 3 and 6.

HOMOLOGY VECTOR 791-63.19. The plasmid 791-63.19 was constructed for the purpose of inserting foreign DNA into SPV. It incorporates an E. coli /S-galactosidase (lacZ) marker gene flanked by SPV DNA. Upstream of the foreign genes is an approximately 1484 base pair fragment of SPV DNA. Downstream of the foreign genes is an approximately 2149 base pair fragment of SPV DNA. When the 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 (LPl) . It was constructed utilizing standard recombinant DNA techniques (22, 30) , by joining restriction fragments from the following sources with the synthetic DNA sequence. The plasmid vector was derived from an approximately 2972 base pair Hindlll to BamHI restriction fragment of pSP64 (Promega) . Fragment 1 is an approximately 1484 base pair Bglll 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 2149 base pair AccI to Hindlll subfragment of the SPV Hindlll fragment M. The AccI sites in fragments 1 and 3 were converted to unique NotI sites using NotI linkers.

HOMOLOGY VECTOR 791-63.41. The plasmid 791-63.41 was constructed for the purpose of inserting foreign DNA into SPV. It incorporates an E. coli _/S-galactosidase (lacZ) marker gene flanked by SPV DNA. Upstream of the foreign genes is an approximately 1484 base pair fragment of SPV DNA. Downstream of the foreign genes is an approximately 2149 base pair fragment of SPV DNA. When the 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 (LP2) . It was constructed utilizing standard recombinant DNA techniques

(22, 30) , by joining restriction fragments from the following sources with the synthetic DNA sequences. The plasmid vector was derived from an approximately 2972 base pair Hindlll to BamHI restriction fragment of pSP64 (Promega) . Fragment 1 is an approximately 1484 base pair Bglll 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 2149 base pair AccI to Hindlll subfragment of the SPV Hindlll fragment M. The AccI sites in fragments 1 and 3 were converted to unique NotI sites using NotI linkers.

HOMOLOGY VECTOR 796-18.9. The plasmid 796-18.9 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 foreign genes is an approximately 1484 base pair fragment of SPV DNA. Downstream of the foreign genes is an approximately 2149 base pair fragment of SPV DNA. When the 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 early pox promoter (EP1) . It was constructed utilizing standard recombinant DNA techniques

(22, 30) , by joining restriction fragments from the following sources with the synthetic DNA sequences. The plasmid vector was derived from an approximately 2972 base pair Hindlll to BamHI restriction fragment of pSP64 (Promega) . Fragment 1 is an approximately 1484 base pair Bglll 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 2149 base pair AccI to Hindlll subfragment of the SPV Hindlll fragment M. The AccI sites in fragments 1 and 3 were converted to unique NotI sites using NotI linkers.

HOMOLOGY VECTOR 783-39.2. The plasmid 783-39.2 was constructed for the purpose of inserting foreign DNA into SPV. It incorporates an E. coli S-galactosidase (lacZ) marker gene and an bovine viral diarrhea virus glycoprotein 53 (BVDV gp53) gene flanked by SPV DNA. Upstream of the foreign gene is an approximately 1484 base pair fragment of SPV DNA. Downstream of the foreign genes is an approximately 2149 base pair fragment of SPV DNA. When the 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 _/S-galactosidase (lacZ) marker gene is under the control of a late promoter (LPl) and the BVDV gp53 gene is under the control of the late/early promoter (LP2EP2) . It was constructed utilizing standard recombinant DNA techniques (22, 30) , by joining restriction fragments from the following sources with the synthetic DNA sequences. The plasmid vector was derived from an approximately 2972 base pair Hindlll to BamHI restriction fragment of pSP64 (Promega) . Fragment 1 is an approximately 1484 base pair Bglll to AccI restriction sub-fragment of the SPV Hindlll restriction fragment M (23) . Fragment 2 is an approximately 1187 base pair BamHI fragment coding for the BVDV gp53. The 1187 base pair BamHI fragment was synthesized by polymerase chain reaction (15) as described in CLONING OF BOVINE VIRAL DIARRHEA VIRUS gp48 AND gp53 GENES. Fragment 3 is an approximately 3010 base pair BamHI to PvuII restriction fragment of plasmid pJF751 (11) . Fragment 4 is an approximately 2149 base pair AccI to Hindlll subfragment of the SPV Hindlll fragment M. The AccI sites in fragments 1 and 4 were converted to unique NotI sites using NotI linkers.

HOMOLOGY VECTOR 749-75.78. The plasmid 749-75.78 was constructed for the purpose of inserting foreign DNA into SPV. It incorporates an E. coli _/S-galactosidase (lacZ) marker gene and the infectious bursal disease virus

(IBDV) polymerase gene flanked by SPV DNA. Upstream of the foreign genes is an approximately 1484 base pair fragment of SPV DNA. Downstream of the foreign genes is an approximately 2149 base pair fragment of SPV DNA. When the 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 (LPl) and the IBDV polymerase gene is under the control of a synthetic late/early promoter (LP2EP2) . It was constructed utilizing standard recombinant DNA techniques

(22, 30) , by joining restriction fragments from the following sources with the synthetic DNA sequences. The plasmid vector was derived from an approximately 2972 base pair Hindlll to BamHI restriction fragment of pSP64

(Promega) . Fragment 1 is an approximately 1484 base pair

Bglll to AccI restriction sub-fragment of the SPV Hindlll restriction fragment M (23) . Fragment 2 is an approximately 2700 EcoRI to Ascl restriction fragment synthesized by cDNA cloning and polymerase chain reaction (PCR) from an IBDV RNA template. cDNA and PCR primers (5' -CACGAATTCTGACATTTTCAACAGTCCACAGGCGC-3'; 12/93.4) (SEQ ID NO: ) and 5' -GCTGTTGGACATCACGGGCCAGG-3' ; 9/93.28) (SEQ ID NO: ) were used to synthesize an approximately 1400 base pair EcoRI to Bell fragment at the 5' end of the IBDV polymerase gene. cDNA and PCR primers (5'- ACCCGGAACATATGGTCAGCTCCAT-3' ; 12/93.2) (SEQ ID NO: ) and 5' -GGCGCGCCAGGCGAAGGCCGGGGATACGG-3' ; 12/93.3) (SEQ ID NO: ) were used to synthesize an approximately 1800 base pair Bell to Ascl fragment at the 3' end of the IBDV polymerase gene. The two fragments were ligated at the Bell site to form the approximately 2700 base pair EcoRI to Bell fragment. Fragment 3 is an approximately 3010 base pair BamHI to PvuII restriction fragment of plasmid pJF751 (11) . Fragment 4 is an approximately 2149 base pair AccI to Hindlll subfragment of the SPV Hindlll fragment M. The AccI sites in fragments 1 and 4 were converted to unique NotI sites using NotI linkers.

HOMOLOGY VECTOR 761-75.B18. The plasmid 761-75.B18 was constructed for the purpose of inserting foreign DNA into SPV. It incorporates an E. coli 0-galactosidase (lac Z) marker gene and a feline immunodeficiency virus (FIV) protease (gag) gene flanked by SPV DNA. Upstream of the foreign gene is an approximately 855 base pair fragment of SPV DNA. Downstream of the foreign genes is an approximately 1113 base pair fragment of SPV DNA. When the 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 swinepox virus OIL gene promoter and the FIV gag gene is under the control of the late/early promoter (LP2EP2) . The homology vector was constructed utilizing standard recombinant DNA techniques (22, 30) , by joining restriction fragments from the following sources with the synthetic DNA sequences. The plasmid vector was derived from an approximately 2519 base pair Hindlll to SphI restriction fragment of pSP65

(Promega) . Fragment 1 is an approximately 855 base pair sub-fragment of the SPV Hindlll restriction fragment M

(23) synthesized by polymerase chain reaction using DNA primers 5' GAAGCATGCCCGTTCTTATCAATAGTTTAGTCGAAAATA-3 '

( S E Q I D N O : 1 8 5 ) a n d 5 ' -

CATAAGATCTGGCATTGTGTTATTATACTAACAAAAATAAG-3' (SEQ ID NO: 186) to produce an 855 base pair fragment with SphI and Bglll ends. Fragment 2 is a 3002 base pair BamHI to PvuII fragment derived from plasmid pJF751 (49) containing the E. coli lacZ gene. Fragment 3 is an approximately 1878 base pair EcoRI to Bglll restriction fragment synthesized by polymerase chain reaction (PCR) using cDNA from the FIV (PPR strain) (61) . The primer (5' GCGTGAATTCGGGGAATGGACAGGGGCGAGAT-3' ; 11/94.9) (SEQ ID NO: ) synthesizes from the 5' end of the FIV gag gene, introduces an EcoRI site at the 5' end of the gene and an ATG start codon. The primer (5' - GAGCCAGATCTGCTCTTTTTACTTTCCC-3' ; 11/94.10) (SEQ ID NO: ) synthesizes from the 3' end of the FIV gag gene. The PCR product was digested with EcoRI and Bglll to yield a fragment 1878 base pairs in length corresponding to the FIV gag gene. Fragment 4 is an approximately 1113 base pair subfragment of the SPV Hindlll fragment M synthesized by polymerase chain reaction using DNA primers 5' -CCGTAGTCGACAAAGATCGACTTATTAATATGTATGGGATT-3 '

( S E Q I D N O : 1 8 7 ) a n d 5 '

GCCTGAAGCTTCTAGTACAGTATTTACGACTTTTGAAAT-3' (SEQ ID NO:

188) to produce an 1113 base pair fragment with Sail and Hindlll ends.

HOMOLOGY VECTOR 781-84.Cll. The plasmid 781-84.Cll was used to insert foreign DNA into SPV. It incorporates an E. coli _/S-galactosidase (lacZ) marker gene and the feline immunodeficiency virus (FIV) envelope (env) gene flanked by SPV DNA. When this plasmid was used according to the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT SPV a virus containing DNA coding for the foreign genes results. Note that the β galactosidase (lacZ) marker gene is under the control of a synthetic late pox promoter (LPl) and the FIV env gene is under the control of a synthetic late/early pox promoter (LP2EP2) . The homology vector was constructed utilizing standard recombinant DNA techniques (22 and 30) , by joining restriction fragments from the following sources with the appropriate synthetic DNA sequences. The plasmid vector was derived from an approximately 2972 base pair Hindlll to BamHI restriction fragment of pSP64 (Promega) . Fragment 1 is an approximately 1484 base pair Bglll to AccI restriction sub-fragment of the SPV Hindlll fragment M (23) . Fragment 3 is an approximately 2564 base pair BamHI to BamHI fragment of the FIV env gene (61) synthesized by CLONING WITH THE POLYMERASE CHAIN REACTION. The template for the PCR reaction was FIV strain PPR genomic cDNA (61) . The upstream primer 10/93.21 (5'-GCCCGGATCCTATGGCAGAAGGGTTTGCAGC-3' ; SEQ ID NO.) was synthesized corresponding to the 5' end of the FIV env gene starting at nucleotide 6263 of FIV strain PPR genomic cDNA, and the procedure introduced a BamHI site at the 5' end. The BamHI site was destroyed during the cloning of the PCR fragment . The downstream primer 10/93.20 (5' -CCGTGGATCCGGCACTCCATCATTCCTCCTC-3' ; SEQ ID NO. ) was synthesized corresponding to the 3' end of the FIV env gene starting at nucleotide 8827 of FIV PPR genomic cDNA, and the procedure introduced a BamHI site at the 3' end. Fragment 3 is an approximately 3010 base pair BamHI to PvuII restriction fragment of plasmid pJF751 (11) . Fragment 4 is an approximately 2149 base pair AccI to Hindlll restriction sub-fragment of the SPV Hindlll restriction fragment M (23) . The AccI site in the SPV homology vector was converted to a unique NotI site.

EXAMPLES

Example 1

Homology 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. AccI site in homology vector 515-85.1 is 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 Figures 1A-1B) 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 jβ-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 /S-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 sequenced. The first region covers a 599 base pair sequence which flanks the unique AccI site (see Figures 2A and 2B) . The second region covers the 899 base pairs upstream of the unique Hindlll si te (see Figures 2A and 2B) . The sequence of the first region codes for an open reading frame (ORF) which shows homology to amino acids 1 to 115 of the vaccinia virus (W) OIL open reading frame identified by Goebel et al , 1990 (see Figures 3A-3C) . 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 Figures 3A-3C) . These da ta sugges t 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 _/S-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 _/S-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 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 _/S-galactosidase (lacZ gene) and the gene for pseudorabies virus (PRV) g50 (gD) (26) were inserted into the SPV 515-85.1 ORF. The lacZ gene is under the control of a synthetic late promoter (LPl) and the g50 (gD) 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 _/S-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 g50 (gD) 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 g50 (gD) (35) .

PRV g50 (gD) is the g50 (gD) homologue of HSV-1 (26) . Several investigators have shown that W expressing HSV-1 g50 (gD) will protect mice against challenge with HSV-1 (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 gll (28) , gill (27) , and gH (19) . The PRV gill 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 gX or gl, 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- 011

S-SPV-011 is a swinepox virus that expresses at least two foreign genes. The gene for E. coli 3-galactosidase

(lacZ) and the gene for pseudorabies virus gill (gC) were inserted into the unique PstI restriction site (PstI linkers inserted into a unique AccI site) of the homology vector 570-33.32. The lac Z gene is under the control of the synthetic late promoter (LPl) and the PRV gill (gC) gene is under the control of the synthetic early promoter

(EP2) .

S-SPV-011 was derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing the homology vector 570- 91.21 (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 _/S-galactosidase

(BLUOGAL AND CPRG ASSAYS) . The final result of red plaque purification was the recombinant virus designated

S-SPV-011. This virus was assayed for _/S-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 foreign gene.

S-SPV-011 was assayed for expression of PRV specific antigens using the BLACK PLAQUE SCREEN FOR FOREIGN GENE EXPRESSION IN RECOMBINANT SPV. Polyclonal goat anti-PRV gill (gC) antibody was shown to react specifically with S-SPV-011 plaques and not with S-SPV-001 negative control plaques. All S-SPV-011 observed plaques reacted with the swine anti-PRV serum indicating that the virus was stably expressing the PRV foreign gene. The assays described here were carried out in EMSK cells, indicating that EMSK cells would be a suitable substrate for the production of SPV recombinant vaccines.

To confirm the expression of the PRV gill (gC) 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. Polyclonal goat anti-PRV gill (gC) antibody was used to detect expression of PRV specific proteins. As shown in Figure 16, the lysate from S-SPV-011 infected cells exhibits a specific band of approximately 92 kd, the reported size of PRV gill (gC) (37) .

Recombinant-expressed PRV gill (gC) has been shown to elicit a significant immune response in mice and swine (37, 38) . Furthermore, when gill (gC) is coexpressed with gll (gB) or g50 (gD) , significant protection from challenge with virulent PRV is obtained (39) . Therefore S-SPV-011 should be valuable as a vaccine to protect swine against PRV disease. Since the PRV vaccines described here do not express PRV gX or gl, 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.

Example 5

S-SPV- 012

S-SPV-012 is a swinepox virus that expresses at least two foreign genes. The gene for E. coli _/β-galactosidase (lacZ) and the gene for pseudorabies virus gill (gC) were inserted into the unique PstI restriction site (PstJ linkers inserted into a unique AccI site) of the homology vector 570-33.32. The lacZ gene is under the control of the synthetic late promoter (LPl) and the PRV gill (gC) gene is under the control of the synthetic early late promoter (EP1LP2) .

S-SPV-012 was derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing the homology vector 570- 91.41 (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 _/S-galactosidase

(BLUOGAL AND CPRG ASSAYS) . The final result of red plaque purification was the recombinant virus designated

S-SPV-012. This virus was assayed for 3-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 foreign gene.

S-SPV-012 was assayed for expression of PRV specific antigens using the BLACK PLAQUE SCREEN FOR FOREIGN GENE EXPRESSION IN RECOMBINANT SPV. Polyclonal goat anti-PRV gill (gC) antibody was shown to react specifically with S-SPV-012 plaques and not with S-SPV-001 negative control plaques. All S-SPV-012 observed plaques reacted with the swine anti-PRV serum, indicating that the virus was stably expressing the PRV foreign gene. The assays described here were carried out in EMSK and VERO cells, indicating that EMSK cells would be a suitable substrate for the production of SPV recombinant vaccines.

To confirm the expression of the PRV gill (gC) gene product, cells were infected with S-SPV-012 and samples of infected cell lysates were subjected to SDS- polyacrylamide gel electrophoresis. The gel was blotted and analyzed using the WESTERN BLOTTING PROCEDURE. Polyclonal goat anti-PRV gill (gC) antibody was used to detect expression of PRV specific proteins. As shown in Figure 16, the lysate from S-SPV-012 infected cells exhibits two specific bands which are the reported size of PRV gill (gC) (37) - a 92 kd mature form and a 74 kd pre-golgi form.

Recombinant-expressed PRV gill (gC) has been shown to elicit a significant immune response in mice and swine (37, 38) . Furthermore, when gill (gC) is coexpressed with gll (gB) or g50 (gD) , significant protection from challenge with virulent PRV is obtained (39) . Therefore S-SPV-012 should be valuable as a vaccine to protect swine against PRV disease. Since the PRV vaccines described here do not express PRV gX or gl, 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.

Example 6

S-SPV- 013

S-SPV-013 is a swinepox virus that expresses at least two foreign genes. The gene for E. coli _/S-galactosidase (lacZ) and the gene for pseudorabies virus gill (gC) were inserted into the unique PstI restriction site (PstI linkers inserted into a unique AccI site) of the homology vector 570-33.32. The lacZ gene is under the control of the synthetic late promoter (LPl) and the PRV gill (gC) gene is under the control of the synthetic late early promoter (LP2EP2) .

S-SPV-013 was derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing the homology vector 570- 91.64 (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 _/8-galactosidase

(BLUOGAL AND CPRG ASSAYS) . The final result of red plaque purification was the recombinant virus designated

S-SPV-013. 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 foreign gene.

S-SPV-013 was assayed for expression of PRV specific antigens using the BLACK PLAQUE SCREEN FOR FOREIGN GENE EXPRESSION IN RECOMBINANT SPV. Polyclonal goat anti-PRV gill (gC) antibody was shown to react specifically with S-SPV-013 plaques and not with S-SPV-001 negative control plaques. All S-SPV-013 observed plaques reacted with the swine anti-PRV serum indicating that the virus was stably expressing the PRV foreign gene. The assays described here were carried out in EMSK and VERO cells, indicating that EMSK cells would be a suitable substrate for the production of SPV recombinant vaccines.

To confirm the expression of the PRV gill (gC) 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. Polyclonal goat anti-PRV gill (gC) antibody was used to detect expression of PRV specific proteins. As shown in Figure 16, the lysate from S-SPV-013 infected cells exhibits two specific bands which are the reported size of PRV gill (gC) (37)-a 92 kd mature form and a 74 kd pre-Golgi form.

Recombinant-expressed PRV gill (gC) has been shown to elicit a significant immune response in mice and swine (37, 38) . Furthermore, when gill (gC) is coexpressed with gll (gB) or g50 (gD) , significant protection from challenge with virulent PRV is obtained. (39) Therefore S-SPV-013 is valuable as a vaccine to protect swine against PRV disease. Since the PRV vaccines described here do not express PRV gX or gl, 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-013 has been deposited with the ATCC under Accession No. 2418.

Protection against Aujeszky's disease using recombinant swinepox virus vaccines S-SPV-008 and S-SPV-013.

A vaccine containing S-SPV-008 and S-SPV-013 (1 x 10⁶PFU/ml) (2ml of a 1:1 mixture of the two viruses) was given to two groups of pigs (5 pigs per group) by intradermal inoculation or by oral/pharyngeal spray. A control group of 5 pigs received S-SPV-001 by both intradermal and oral/pharyngeal inoculation. Pigs were challenged three weeks post-vaccination with virulent PRV, strain 4892, by intranasal inoculation. The table presents a summary of clinical responses. The data support an increase in protection against Aujeszky's disease in the S-SPV-008/S-SPV-013 vaccinates compared to the S-SPV-001 vaccinate controls.

Vaccine Route of Post- Post- Post-challen inoculation challenge challenge Group averag Respiratory- CNS signs: (Days of Signs: (# with clinical (# with signs/ signs) signs/ total number) total number)

S-SPV-008 + Intradermal 3/5 0/5 2.6 S-SPV-013

S-SPV-008 + Oral/ 3/5 0/5 2.2 S-SPV-013 pharyngeal

S-SPV-001 Intradermal + 5/5 2/5 7.8 (Control) Oral/

Pharyngeal

Example 7

S-SPV-015

S-SPV-015 is a swinepox virus that expresses at least two foreign genes. The gene for E. coli S-galactosidase (lacZ) and the gene for pseudorabies virus (PRV) gll (gB) were inserted into the SPV 617-48.1 ORF (a unique NotI restriction site has replaced a unique AccI restriction site) . The lacZ gene is under the control of the synthetic late promoter (LPl) , and the PRV gB gene is under the control of the synthetic late/early promoter (LP2EP2) . S-SPV-015 was derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing the homology vector 727-54.60

(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 _/8-galactosidase

(BLUOGAL AND CPRG ASSAYS) . The final result of red plaque purification was the recombinant virus designated S-SPV-

015. 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 foreign gene.

S-SPV-015 was assayed for expression of PRV-specific antigens using the BLACK PLAQUE SCREEN FOR FOREIGN GENE EXPRESSION IN RECOMBINANT SPV. Polyclonal swine anti-PRV serum was shown to react specifically with S-SPV-015 plaques and not with S-SPV-001 negative control plaques. All S-SPV-015 observed plaques reacted with the antiserum indicating that the virus was stably expressing the PRV foreign gene. The assays described here were carried out in ESK-4 cells, indicating that ESK-4 cells would be a suitable substrate for the production of SPV recombinant vaccines.

To confirm the expression of the PRV gll gene product, cells were infected with SPV-015 and samples of infected cell lysates were subjected to SDS-polyacrylamide gel electrophoresis. The gel was blotted and analyzed using the WESTERN BLOTTING PROCEDURE. Polyclonal swine anti-PRV serum was used to detect expression of PRV specific proteins. The lysate from S-SPV-015 infected cells exhibited bands corresponding to 120 kd, 67 kd and 58 kd, which are the expected size of the PRV gll glycoprotein. S-SPV-015 is useful as a vaccine in swine against pseudorabies virus. A superior vaccine is formulated by combining S-SPV-008 (PRV g50) , S-SPV-013 (PRV glll), and S-SPV-015 for protection against pseudorabies in swine.

Therefore S-SPV-015 should be valuable as a vaccine to protect swine against PRV disease. Since the PRV vaccines described here do not express PRV gX or gl, 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-015 has been deposited with the ATCC under Accession No. 2466.

Example 8

Recombinant swinepox virus expressing more than one pseudorabies virus (PRV) glycoproteins, which can elicit production of neutralizing antibodies against pseudorabies virus, is constructed in order to obtain a recombinant swinepox virus with enhanced ability to protect against PRV infection than that which can be obtained by using a recombinant swinepox virus expressing only one of those PRV glycoproteins .

There are several examples of such recombinant swinepox virus expressing more than one PRV glycoproteins: a recombinant swinepox virus expressing PRV g50 (gD) and gill (gC) , a recombinant swinepox virus expressing PRV g50 (gD) and gll (gB) ,- a recombinant swinepox virus expressing PRV gll (gB) and gill (gC) ; and a recombinant swinepox virus expressing PRV g50 (gD) , gill (gC) and gll (gB) . Each of the viruses cited above is also engineered to contain and express E. coli -galactosidase (lac Z) gene, which will facilitate the cloning of the recombinant swinepox virus. Listed below are three examples of a recombinant swinepox virus expressing PRV g50 (gD) , PRV gill (gC) , PRV gll (gB) and E. coli /8-galactosidase (lacZ) :

a) Recombinant swinepox virus containing and expressing PRV g50 (gD) gene, PRV gill (gC) gene, PRV gll (gB) gene and lacZ gene. All four genes are inserted into the unique AccI restriction endonuclease site within the Hindlll M fragment of the swinepox virus genome. PRV g50 (gD) gene is under the control of a synthetic early/late promoter

(EP1LP2) , PRV gill (gC) gene is under the control of a synthetic early promoter (EP2) , PRV gll (gB) gene is under the control of a synthetic late/early promoter (LP2EP2) and lacZ gene is under the control of a synthetic late promoter (LPl) .

b) Recombinant swinepox virus containing and expressing PRV g50 (gD) gene, PRV gill (gC) gene, PRV gll (gB) gene and lacZ gene. All four genes are inserted into the unique AccI restriction endonuclease site within the Hindlll M fragment of the swinepox virus genome. PRV g50 (gD) gene is under the control of a synthetic early/late promoter (EP1LP2) , PRV gill (gC) gene is under the control of a synthetic early/late promoter (EP1LP2) , PRV gll (gB) gene is under the control of a synthetic late/early promoter (LP2EP2) and lacZ gene is under the control of a synthetic late promoter (LPl) .

c) Recombinant swinepox virus containing and expressing PRV g50 (gD) gene, PRV gill (gC) gene, PRV gll (gB) gene and lacZ gene. All four genes are inserted into the unique AccI restriction endonuclease site within the Hindlll M fragment of the swinepox virus genome. PRV g50 (gD) gene is under the control of a synthetic early/late promoter (EP1LP2), PRV gill (gC) gene is under the control of a synthetic late/early promoter (LP2EP2) , PRV gll (gB) gene is under the control of a synthetic late/early promoter (LP2EP2) and lacZ gene is under the control of a synthetic late promoter (LPl) .

Protection against Aujeszky's disease using recombinant swinepox virus vaccines S-SPV-008, S-SPV-013 and S-SPV- 015.

A vaccine containing S-SPV-008, S-SPV-013, or S-SPV-015 (2 ml of 1 X 10⁷ PFU/ml of the virus) or a mixture of S- SPV-008, S-SPV-013, and S-SPV-015 (2ml of a 1:1:1 mixture of the three viruses; 1 X 10⁷ PFU/ml) was given to four groups of pigs (5 pigs per group) by intramuscular inoculation. A control group of 5 pigs received S-SPV-001 by intramuscular inoculation. Pigs were challenged four weeks post-vaccination with virulent PRV, strain 4892, by intranasal inoculation. The pigs were observed daily for 14 days for clinical signs of pseudorabies, and the table presents a summary of clinical responses.The data show that pigs vaccinated with S-SPV-008, S-SPV-013, or S-SPV- 015 had partial protection and pigs vacinated with the combination vaccine S-SPV-008/S-SPV-013/S-SPV-015 had complete protection against Aujeszky's disease caused by pseudorabies virus compared to the S-SPV-001 vaccinate controls.

Vaccine Route of Post- Post- Post- inoculation challenge challenge challenge Respiratory CNS signs : Group Signs: (# with average : (# with signs/ (Days of signs/ total number) clinical total signs) number)

S-SPV-008 Intramuscular 2/5 2/5 2.0

Example 9

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 (LPl) 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 _/S-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 -S-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 Accession 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 10

S-SPV-014

S-SPV-014 is a swinepox virus that expresses at least two foreign genes. The gene for E. coli β- galactosidase (lacZ) and the gene for infectious laryngotracheitis virus glycoprotein G (ILT gG) were inserted into the SPV 570-33.32 ORF (a unique PstI site has replaced the unique AccI site) . The lacZ gene is under the control of the synthetic late promoter (LPl) , and the ILT gG gene is under the control of the synthetic early/late promoter (EP1LP2) .

S-SPV-014 was derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing the homology vector 599-65.25 (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-014. 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 foreign gene. The assays described here were carried out in ESK-4 cells, indicating that ESK-4 cells would be a suitable substrate for the production of SPV recombinant vaccines.

To confirm the expression of the ILT gG gene product, cells were infected with SPV-014 and samples of infected cell lysates were subjected to SDS- polyacrylamide gel electrophoresis. The gel was blotted and analyzed using the WESTERN BLOTTING

PROCEDURE. Peptide antisera to ILT gG was used to detect expression of ILT specific proteins. The lysate from S-SPV-014 infected cells exhibited a band at 43 kd which is the expected size of the ILT gG protein and additional bands of higher molecular weight which represent glycosylated forms of the protein which are absent in deletion mutants for ILT gG.

This virus is used as an expression vector for expressing ILT glycoprotein G (gG) . Such ILT gG is used as an antigen to identify antibodies directed against the wild-type ILT virus as opposed to antibodies directed against gG deleted ILT viruses. This virus is also used as an antigen for the production of ILT gG specific monoclonal antibodies. Such antibodies are useful in the development of diagnostic tests specific for the ILT gG protein.

Monoclonal antibodies are generated in mice utilizing this virus according to the PROCEDURE FOR PURIFICATION

OF VIRAL GLYCOPROTEINS FOR USE AS DIAGNOSTICS

(Materials & Methods) .

Example 11

S-SPV-016

S-SPV-016 is a swinepox virus that expresses at least two foreign genes. The gene for E. coli β- galactosidase (lacZ) and the gene for infectious laryngotracheitis virus glycoprotein I (ILT gl) were inserted into the SPV 617-48.1 ORF (a unique NotI restriction site has replaced a unique AccI restriction site) . The lacZ gene is under the control of the synthetic late promoter (LPl) , and the ILT gl gene is under the control of the synthetic late/early promoter (LP2EP2) .

S-SPV-016 was derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing the homology vector

624-20.1C (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 _/S-galactosidase (BLUOGAL AND CPRG ASSAYS) . The final result of red plaque purification was the recombinant virus designated S-SPV-016. This virus was assayed for _/S-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 foreign gene.

S-SPV-016 was assayed for expression of ILT gl- and β- galactosidase-specific antigens using the BLACK PLAQUE SCREEN FOR FOREIGN GENE EXPRESSION IN RECOMBINANT SPV. Polyclonal chicken anti-ILT antibody was shown to react specifically with S-SPV-016 plaques and not with S-SPV- 017 negative control plaques. All S-SPV-016 observed plaques reacted with the chicken antiserum indicating that the virus was stably expressing the ILT foreign gene. The assays described here were carried out in ESK-4 cells, indicating that ESK-4 cells would be a suitable substrate for the production of SPV recombinant vaccines.

To confirm the expression of the ILT gl gene product, cells were infected with SPV-016 and samples of infected cell lysates were subjected to SDS- polyacrylamide gel electrophoresis. The gel was blotted and analyzed using the WESTERN BLOTTING PROCEDURE. Polyclonal chicken anti-ILT antibody was used to detect expression of ILT specific proteins. The lysate from S-SPV-016 infected cells exhibits a range of bands reactive to the anti-ILT antibody from 40 to 200 kd indicating that the ILT gl may be heavily modified.

This virus is used as an expression vector for expressing ILT glycoprotein I (gl) . Such ILT gl is used as an antigen to identify antibodies directed against the wild-type ILT virus as opposed to antibodies directed against gl deleted ILT viruses. This virus is also used as an antigen for the production of ILT gl specific monoclonal antibodies. Such antibodies are useful in the development of diagnostic tests specific for the ILT gl protein. Monoclonal antibodies are generated in mice utilizing this virus according to the PROCEDURE FOR PURIFICATION

OF VIRAL GLYCOPROTEINS FOR USE AS DIAGNOSTICS

(Materials & Methods) .

Example 12

S-SPV-017

S-SPV-017 is a swinepox virus that expresses at least two foreign genes. The gene for E. coli β- galactosidase (lacZ) and the gene for infectious bovine rhinotracheitis virus glycoprotein G (IBR gG) were inserted into the SPV 617-48.1 ORF (a unique NotI restriction site has replaced a unique AccI restriction site) . The lacZ gene is under the control of the synthetic late promoter (LPl) , and the IBR gG gene is under the control of the synthetic late/early promoter

(LP2EP2) .

S-SPV-017 was derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing the homology vector 614-83.18 (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 _/8-galactosidase (BLUOGAL AND CPRG ASSAYS) . The final result of red plaque purification was the recombinant virus designated S-SPV-017. This virus was assayed for S-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 foreign gene. S-SPV-017 was assayed for expression of IBR-specific antigens using the BLACK PLAQUE SCREEN FOR FOREIGN GENE EXPRESSION IN RECOMBINANT SPV. Monoclonal antibodies and peptide antisera to IBR gG were shown to react specifically with S-SPV-017 plaques and not with S-SPV- 016 negative control plaques. All S-SPV-017 observed plaques reacted with the antiserum indicating that the virus was stably expressing the IBR foreign gene. The assays described here were carried out in ESK-4 cells, indicating that ESK-4 cells would be a suitable substrate for the production of SPV recombinant vaccines.

To confirm the expression of the IBR gG gene product, cells were infected with SPV-017 and samples of infected cell lysates were subjected to SDS- polyacrylamide gel electrophoresis. The gel was blotted and analyzed using the WESTERN BLOTTING PROCEDURE. Antisera to IBR gG was used to detect expression of IBR specific proteins. The lysate from S-SPV-017 infected cells exhibited a band at 43 kd which is the expected size of the IBR gG protein and additional bands of higher molecular weight which represent glycosylated forms of the protein which are absent in deletion mutants for IBR gG.

This virus is used as an expression vector for expressing IBR glycoprotein G (gG) . Such IBR gG is used as an antigen to identify antibodies directed against the wild-type IBR virus as opposed to antibodies directed against gG deleted IBR viruses. This virus is also used as an antigen for the production of IBR gG specific monoclonal antibodies. Such antibodies are useful in the development of diagnostic tests specific for the IBR gG protein. Monoclonal antibodies are generated in mice utilizing this virus according to the PROCEDURE FOR PURIFICATION OF VIRAL GLYCOPROTEINS FOR USE AS DIAGNOSTICS (Materials & Methods) .

Example 13

S-SPV-019

S-SPV-019 is a swinepox virus that expresses at least two foreign genes. The gene for E. coli _/β-galactosidase (lacZ) and the gene for infectious bovine rhinotracheitis virus (IBRV) gE were inserted into the SPV 617-48.1 ORF (a unique NotI restriction site has replaced a unique AccI restriction site) . The lacZ gene is under the control of the synthetic late promoter (LPl) , and the IBRV gE gene is under the control of the synthetic late/early promoter (LP2EP2) .

S-SPV-019 was derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing the homology vector 708-78.9 (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 /3-galactosidase (BLUOGAL AND CPRG ASSAYS) . The final result of red plaque purification was the recombinant virus designated S-SPV-019. This virus was assayed for _/S-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 foreign gene.

This virus is used as an expression vector for expressing IBR glycoprotein E (gE) . Such IBR gE is used as an antigen to identify antibodies directed against the wild-type IBR virus as opposed to antibodies directed against gE deleted IBR viruses. This virus is also used as an antigen for the production of IBR gE specific monoclonal antibodies. Such antibodies are useful in the development of diagnostic tests specific for the IBR gE protein. Monoclonal antibodies are generated in mice utilizing this virus according to the PROCEDURE FOR PURIFICATION OF VIRAL GLYCOPROTEINS FOR USE AS DIAGNOSTICS (Materials & Methods) .

Example 14

S-SPV-018

S-SPV-018 is a swinepox virus that expresses at least two foreign genes. The gene for E. coli β- galactosidase (lacZ) and the gene for pseudorabies virus glycoprotein E (PRV gE) are inserted into the SPV 570-33.32 ORF (a unique PstI site has replaced the unique AccI site) . The lacZ gene is under the control of the synthetic late promoter (LPl) , and the PRV gE gene is under the control of the synthetic early/late promoter (EP1LP2) .

S-SPV-018 is derived from the S-SPV-001 (Kasza Strain) . This is accomplished utilizing the final homology vector and virus S-SPV-001 in the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT SPV. The transfection stock is screened by the SCREEN FOR RECOMBINANT SPV EXPRESSING _/S-galactosidase (BLUOGAL AND CPRG ASSAYS) . Red plaque purification of the recombinant virus is designated S-SPV-018. This virus is assayed for /S-galactosidase expression, purity, and insert stability by multiple passages monitored by the blue plaque assay described in Materials and Methods. After the initial three rounds of purification, all plaques observed are blue indicating that the virus is pure, stable, and expressing the foreign gene.

This virus is used as an expression vector for expressing PRV glycoprotein E (gE) . Such PRV gE is used as an antigen to identify antibodies directed against the wild-type PRV virus as opposed to antibodies directed against gE deleted PRV viruses. This virus is also used as an antigen for the production of PRV gE specific monoclonal antibodies. Such antibodies are useful in the development of diagnostic tests specific for the PRV gE protein. Monoclonal antibodies are generated in mice utilizing this virus according to the PROCEDURE FOR PURIFICATION OF VIRAL GLYCOPROTEINS FOR USE AS DIAGNOSTICS (Materials & Methods) .

Example 15

Homolocr^*/ 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 DΝA may be cloned. A plasmid containing such a foreign DΝA 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, thymidine kinase gene of swinepox virus was shown to be non-essential for DNA replication and is an appropriate insertion site.

Example 16

S-PRV-010

S-SPV-010 is a swinepox virus that expresses a foreign gene. The E. coli _/8-galactosidase (lacZ) gene is inserted into a unique Ndel restriction site within the thymidine kinase gene. The foreign gene (lacZ) is under the control of the synthetic late promoter, LPl. Thus, swinepox virus thymidine kinase gene was shown to be non-essential for replication of the virus and is an appropriate insertion site.

A 1739 base pair Hindlll-Ba HI fragment subcloned from the Hindlll G fragment contains the swinepox virus thymidine kinase gene and is designated homology vector 520-90.15. The homology vector 520-90.15 was digested with Nde I, and Ascl linkers were inserted at this unique site within the thymidine kinase gene. The LPl promoter-lac Z cassette with Ascl linkers was ligated into the Asc I site within the thymidine kinase gene. The recombinant homology vector 561-36.26 was cotransfeeted with virus S-SPV-001 by the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT SPV and virus plaques expressing _/β-galactosidase were selected by SCREEN FOR RECOMBINANT SPV EXPRESSING β- galactosidase (BLUOGAL AND CPRG ASSAY) . The final result of blue and red plaque purification was the recombinant virus designated S-SPV-010. This virus was assayed for _/8-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 foreign gene. The assays described here were carried out in ESK-4 cells, indicating that ESK-4 cells would be a suitable substrate for the production of SPV recombinant vaccines.

Example 17

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 g50 (gD) in SPV and are disclosed herein are applicable to TGE.

PORCINE PARVOVIRUS

The major capsid protein of the porcine (swine) parvovirus (PPV) was cloned 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 g50 (gD) in SPV and are disclosed herein are applicable to PPV.

SWINE ROTAVIRUS

The major neutralizing antigen of the swine rotavirus, glycoprotein 38, was cloned 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 g50 (gD) 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 hog cholera viruses are cross protective (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 g50 (gD) in SPV and are disclosed herein are applicable to BVD virus.

SERPULINA HYODYSENTERIAE

A protective antigen of Serpulina hyodys enter iae (3), for use in the swinepox virus vector has been cloned. It is contemplated that the procedures that have been used to express PRV g50 in SPV and are disclosed herein are also applicable to Serpulina hyodys enter iae .

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, Mycoplasma hyopneumoniae, porcine reproductive and respiratory syndrome/swine infertility and respiratory syndrome (PRRS/SIRS) .

Antigens from the following microorganisms may also be utilized for animal vaccines: 1) canine - herpesvirus, canine distemper, canine adenovirus type 1 (hepatitis) , adenovirus type 2 (respiratory disease) , parainfluenza, Leptospira canicola, icterohemorragia, parvovirus, coronavirus, Borrelia burgdorferi , canine herpesvirus, Bordetella bronchiseptica, Dirofilaria immi tis (heartworm) and rabies virus. 2) Feline - Fiv gag and env, feline leukemia virus, feline immunodeficiency virus, feline herpesvirus, feline infectious peritonitis virus, canine herpesvirus, canine coronavirus, canine parvovirus, parasitic diseases in animals (including Dirofilaria immi tis in dogs and cats) , equine infectious anemia, Streptococcus equi , coccidia, emeria, chicken anemia virus, Borrelia bergdorferi , bovine coronavirus, Pasteurel la haemolytica .

Example 17A

Vaccines containing recombinant swinepox virus expressing antigens from hog cholera virus, swine influenza virus and (porcine reproducting and respiratory syndrome) PRRS virus.

Recombinant swinepox virus expressing genes for neutralizing antigens to hog cholera virus, swine influenza virus and PRRS virus is useful to prevent disease in swine. The genes expressed in the recombinant SPV include, but are not limited to hog cholera virus gEl and gE2 genes, swine influenza virus hemagglutinin, neuraminidase, matrix and nucleoprotein, and PRRS virus 0RF7.

Example 18

Recombinant swinepox viruses express equine influenza virus type A/Alaska 91, equine influenza virus type

A/Prague 56, equine herpesvirus type 1 gB, or equine herpesvirus type 1 gD genes. S-SPV-033 and S-SPV-034 are useful as vaccines against equine influenza infection, and S-SPV-038 and S-SPV-039 are useful as a vaccine against equine herpesvirus infection which causes equine rhinotracheitis and equine abortion. These equine influenza and equine herpesvirus antigens are key to raising a protective immune response in the animal . The recombinant viruses are useful alone or in combination as an effective vaccine. The swinepox virus is useful for cloning other subtypes of equine influenza virus (including equine influenza virus type A/Miami/63 and equine influenza virus type A/Kentucky/81) to protect against rapidly evolving variants in this disease. S-SPV-033, S-SPV-034, S-SPV- 038, and S-SPV-039 are also useful as an expression vector for expressing equine influenza or equine herpesvirus antigens. Such equine influenza or equine herpesvirus antigens are useful to identify antibodies directed against the wild-type equine influenza virus or equine herpesvirus. The viruses are also useful to in producing antigens for the production of monospecific polyclonal or monoclonal antibodies. Such antibodies are useful in the development of diagnostic tests specific for the viral proteins. Monoclonal or polyclonal antibodies are generated in mice utilizing these viruses according to the PROCEDURE FOR PURIFICATION OF VIRAL GLYCOPROTEINS FOR USE AS DIAGNOSTICS (Materials and Methods) .

Example 18A

S-SPV-033:

S-SPV-033 is a recombinant swinepox virus that expresses at least two foreign genes. The gene for E. coli 3-galactosidase (lacZ) and the gene for equine influenza virus type A/Alaska 91 neuraminidase were inserted into the SPV 617-48.1 ORF (a unique NotI restriction site has replaced a unique AccI restriction site) . The lacZ gene is under the control of the synthetic late promoter (LPl), and the EIV AK/91 ΝA gene is under the control of the synthetic late/early promoter (LP2EP2) . S-SPV-033 was derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing the homology vector 732-18.4 (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 /S-galactosidase (BLUOGAL AND CPRG ASSAYS) . The final result of red plaque purification was the recombinant virus designated S-SPV-033. This virus was assayed for /S-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 foreign gene.

Example 18B

S-SPV-034

S-SPV-034 is a swinepox virus that expresses at least two foreign genes. The gene for E. coli _/S-galactosidase

(lacZ) and the gene for equine influenza virus type

A/Prague 56 neuraminidase were inserted into the SPV 617-48.1 ORF (a unique NotI restriction site has replaced a unique AccI restriction site) . The lacZ gene is under the control of the synthetic late promoter (LPl) , and the EIV PR/56 ΝA gene is under the control of the synthetic late/early promoter (LP2EP2) .

S-SPV-034 was derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing the homology vector 723-59A9.22 (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-034. This virus was assayed for _/8-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 foreign gene.

S-SPV-034 was assayed for expression of ElV-specific antigens using the BLACK PLAQUE SCREEN FOR FOREIGN GENE EXPRESSION IN RECOMBINANT SPV. Monospecific polyclonal antibodies to EIV PR/56 NA were shown to react specifically with S-SPV-034 plaques and not with S-SPV- 001 negative control plaques. All S-SPV-034 observed plaques reacted with the antiserum indicating that the virus was stably expressing the EIV PR/56 NA gene. The assays described here were carried out in ESK-4 cells, indicating that ESK-4 cells would be a suitable substrate for the production of SPV recombinant vaccines.

Example 18C

S-SPV-038:

S-SPV-038 is a swinepox virus that expresses at least two foreign genes. The gene for E. coli _/3-galactosidase (lacZ) and the gene for equine herpesvirus type 1 glycoprotein B are inserted into the SPV 617-48.1 ORF

(a unique NotI restriction site has replaced a unique

AccI restriction site) . The lacZ gene is under the control of the synthetic late promoter (LPl) , and the

EHV-1 gB gene is under the control of the synthetic late/early promoter (LP2EP2) . S-SPV-038 is derived from S-SPV-001 (Kasza Strain) . This is accomplished utilizing the homology vector 744- 34 (see Materials and Methods) and virus S-SPV-001 in the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT SPV. The transfection stock is screened by the SCREEN FOR RECOMBINANT SPV EXPRESSING β- galactosidase (BLUOGAL AND CPRG ASSAYS) . The final result of red plaque purification is the recombinant virus designated S-SPV-038. This virus is assayed for /S-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 are blue indicating that the virus is pure, stable, and expressing the foreign gene.

Example 18D

S-SPV-039

S-SPV-039 is a swinepox virus that expresses at least two foreign genes. The gene for E. coli _/S-galactosidase

(lacZ) and the gene for equine herpesvirus type 1 glycoprotein D are inserted into the SPV 617-48.1 ORF (a unique NotI restriction site has replaced a unique AccI restriction site) . The lacZ gene is under the control of the synthetic late promoter (LPl) , and the EHV-1 gD gene is under the control of the synthetic late/early promoter (LP2EP2) .

S-SPV-039 is derived from S-SPV-001 (Kasza Strain) . This is accomplished utilizing the homology vector 744- 38 (see Materials and Methods) and virus S-SPV-001 in the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT SPV. The transfection stock is screened by the SCREEN FOR RECOMBINANT SPV EXPRESSING β- galactosidase (BLUOGAL AND CPRG ASSAYS) . The final result of red plaque purification is the recombinant virus designated S-SPV-039. This virus is assayed for _/S-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 are blue indicating that the virus is pure, stable, and expressing the foreign gene.

Example 19

Recombinant swinepox viruses express bovine respiratory syncytial virus attachment protein (BRSV G) , BRSV Fusion protein (BRSV F) , BRSV nucleocapsid protein (BRSV N) , bovine viral diarrhea virus (BVDV) g48, BVDV g53, bovine parainfluenza virus type 3 (BPI-3) F, or BPI-3 HN. S-SPV-020, S-SPV-029, S-SPV-030, and S-SPV- 032, S-SPV-028 are useful as vaccines against bovine disease. These BRSV, BVDV, and BPI-3 antigens are key to raising a protective immune response in the animal. The recombinant viruses are useful alone or in combination as an effective vaccine. The swinepox virus is useful for cloning other subtypes of BRSV, BVDV, and BPI-3 to protect against rapidly evolving variants in this disease. S-SPV-020, S-SPV-029, S-SPV-030, and S- SPV-032, S-SPV-028 are also useful as an expression vector for expressing BRSV, BVDV, and BPI-3 antigens. Such BRSV, BVDV, and BPI-3 antigens are useful to identify antibodies directed against the wild-type BRSV, BVDV, and BPI-3. The viruses are also useful as antigens for the production of monospecific polyclonal or monoclonal antibodies. Such antibodies are useful in the development of diagnostic tests specific for the viral proteins. Monoclonal or polyclonal antibodies are generated in mice utilizing these viruses according to the PROCEDURE FOR PURIFICATION OF VIRAL GLYCOPROTEINS FOR USE AS DIAGNOSTICS (Materials and Methods) .

Example 19A

S-SPV-020:

S-SPV-020 is a swinepox virus that expresses at least two foreign genes. The gene for E. coli /S-galactosidase

(lacZ) and the gene for bovine respiratory syncytial virus (BRSV) G were inserted into the SPV 617-48.1 ORF (a unique NotI restriction site has replaced a unique

AccI restriction site) . The lacZ gene is under the control of the synthetic late promoter (LPl) , and the

BRSV G gene is under the control of the synthetic late/early promoter (LP2EP2) .

S-SPV-020 was derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing the homology vector 727-20.5 (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 /S-galactosidase (BLUOGAL AND CPRG ASSAYS) . The final result of red plaque purification was the recombinant virus designated S-SPV-020. 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 foreign gene.

S-SPV-020 was assayed for expression of BRSV-specific antigens using the BLACK PLAQUE SCREEN FOR FOREIGN GENE

EXPRESSION IN RECOMBINANT SPV. Bovine anti-BRSV FITC

(Accurate Chemicals) was shown to react specifically with S-SPV-020 plaques and not with S-SPV-003 negative control plaques. All S-SPV-020 observed plaques reacted with the antiserum indicating that the virus was stably expressing the BRSV foreign gene. The assays described here were carried out in ESK-4 cells, indicating that

ESK-4 cells would be a suitable substrate for the production of SPV recombinant vaccines.

To confirm the expression of the BRSV G gene product, cells were infected with S-SPV-020 and samples of infected cell lysates were subjected to SDS- polyacrylamide gel electrophoresis. The gel was blotted and analyzed using the WESTERN BLOTTING PROCEDURE. Bovine anti-BRSV FITC (Accurate Chemicals) was used to detect expression of BRSV specific proteins. The lysate from S-SPV-020 infected cells exhibited a band at 36 kd which is the expected size of the non-glycosylated form of BRSV G protein and bands at 43 to 45 kd and 80 to 90 kd which are the expected size of glycosylated forms of the BRSV G protein.

Example 19B

S-SPV-029

S-SPV-029 is a swinepox virus that expresses at least two foreign genes. The gene for E. coli _/S-galactosidase

(lacZ) and the gene for bovine respiratory syncytial virus (BRSV) F were inserted into the SPV 617-48.1 ORF (a unique NotI restriction site has replaced a unique AccI restriction site) . The lacZ gene is under the control of the synthetic late promoter (LPl) , and the BRSV F gene is under the control of the synthetic late/early promoter (LP2EP2) .

S-SPV-029 was derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing the homology vector 727-20.10 (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-029. This virus was assayed for /S-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 foreign gene.

S-SPV-029 was assayed for expression of BRSV-specific antigens using the BLACK PLAQUE SCREEN FOR FOREIGN GENE

EXPRESSION IN RECOMBINANT SPV. Bovine anti-BRSV FITC

(Accurate Chemicals) was shown to react specifically with S-SPV-029 plaques and not with S-SPV-003 negative control plaques. All S-SPV-029 observed plaques reacted with the antiserum indicating that the virus was stably expressing the BRSV foreign gene. The assays described here were carried out in ESK-4 cells, indicating that ESK-4 cells would be a suitable substrate for the production of SPV recombinant vaccines.

Example 19C

S-SPV-030

S-SPV-030 is a swinepox virus that expresses at least two foreign genes. The gene for E. coli /S-galactosidase

(lacZ) and the gene for bovine respiratory syncytial virus (BRSV) N were inserted into the SPV 617-48.1 ORF (a unique NotI restriction site has replaced a unique AccI restriction site) . The lacZ gene is under the control of the synthetic late promoter (LPl) , and the BRSV Ν gene is under the control of the synthetic late/early promoter (LP2EP2) .

S-SPV-030 was derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing the homology vector 713-55.37 (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 _/S-galactosidase (BLUOGAL AND CPRG ASSAYS) . The final result of red plaque purification was the recombinant virus designated S-SPV-030. This virus was assayed for 3-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 foreign gene.

S-SPV-030 was assayed for expression of BRSV-specific antigens using the BLACK PLAQUE SCREEN FOR FOREIGN GENE

EXPRESSION IN RECOMBINANT SPV. Bovine anti-BRSV FITC

(Accurate Chemicals) was shown to react specifically with S-SPV-030 plaques and not with S-SPV-003 negative control plaques. All S-SPV-030 observed plaques reacted with the antiserum indicating that the virus was stably expressing the BRSV foreign gene. The assays described here were carried out in ESK-4 cells, indicating that ESK-4 cells would be a suitable substrate for the production of SPV recombinant vaccines.

To confirm the expression of the BRSV N gene product, cells were infected with SPV-030 and samples of infected cell lysates were subjected to SDS- polyacrylamide gel electrophoresis. The gel was blotted and analyzed using the WESTERN BLOTTING PROCEDURE. Bovine anti-BRSV FITC (Accurate Chemicals) was used to detect expression of BRSV specific proteins. The lysate from S-SPV-030 infected cells exhibited a band at 43 kd which is the expected size of the BRSV N protein.

Example 19D

S-SPV-028:

S-SPV-028 is a swinepox virus that expresses at least two foreign genes. The gene for E. coli _/S-galactosidase (lacZ) and the gene for bovine parainfluenza virus type 3 (BPI-3) F were inserted into the SPV 617-48.1 ORF (a unique NotI restriction site has replaced a unique AccI restriction site) . The lacZ gene is under the control of the synthetic late promoter (LPl) , and the BPI-3 F gene is under the control of the synthetic late/early promoter (LP2EP2) .

S-SPV-028 was derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing the homology vector 713-55.10 (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 _/S-galactosidase (BLUOGAL AND CPRG ASSAYS) . The final result of red plaque purification was the recombinant virus designated S-SPV-028. This virus was assayed for _/8-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 foreign gene.

S-SPV-028 was assayed for expression of BPI-3-specific antigens using the BLACK PLAQUE SCREEN FOR FOREIGN GENE

EXPRESSION IN RECOMBINANT SPV. Bovine anti-BPI-3 FITC

(Accurate Chemicals) was shown to react specifically with S-SPV-028 plaques and not with S-SPV-003 negative control plaques. All S-SPV-028 observed plaques reacted with the antiserum indicating that the virus was stably expressing the BPI-3 foreign gene. The assays described here were carried out in ESK-4 cells, indicating that ESK-4 cells would be a suitable substrate for the production of SPV recombinant vaccines. To confirm the expression of the BPI-3 F gene product, cells were infected with SPV-028 and samples of infected cell lysates were subjected to SDS- polyacrylamide gel electrophoresis. The gel was blotted and analyzed using the WESTERN BLOTTING PROCEDURE. Bovine anti-BPI-3 FITC (Accurate Chemicals) was used to detect expression of BPI-3 specific proteins. The lysate from S-SPV-028 infected cells exhibited bands at 43, and 70 kd which is the expected size of the BPI-3 F protein.

Example 19E

S-SPV-032

S-SPV-032 is a swinepox virus that expresses at least two foreign genes. The gene for E. coli _/S-galactosidase (lacZ) and the gene for bovine viral diarrhea virus (BVDV) g48 were inserted into the SPV 617-48.1 ORF (a unique NotI restriction site has replaced a unique AccI restriction site) . The lacZ gene is under the control of the synthetic late promoter (LPl) , and the BVDV g48 gene is under the control of the synthetic late/early promoter (LP2EP2) .

S-SPV-032 was derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing the homology vector 727-78.1 (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 _/S-galactosidase (BLUOGAL AND CPRG ASSAYS) . The final result of red plaque purification was the recombinant virus designated S-SPV-032. This virus was assayed for _/S-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 foreign gene.

Example 19F

S-SPV-040:

S-SPV-040 is a swinepox virus that expresses at least two foreign genes. The gene for E. coli _/β-galactosidase

(lacZ) and the gene for bovine viral diarrhea virus

(BVDV) g53 were inserted into the SPV 617-48.1 ORF (a unique NotI restriction site has replaced a unique AccI restriction site) . The lacZ gene is under the control of the synthetic late promoter (LPl) , and the BVDV g53 gene is under the control of the synthetic late/early promoter (LP2EP2) .

S-SPV-040 is derived from S-SPV-001 (Kasza Strain) . This is accomplished utilizing the homology vector 738- 96 (see Materials and Methods) and virus S-SPV-001 in the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT SPV. The transfection stock is screened by the SCREEN FOR RECOMBINANT SPV EXPRESSING β- galactosidase (BLUOGAL AND CPRG ASSAYS) . The final result of red plaque purification is the recombinant virus designated S-SPV-040. This virus is assayed for _/S-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 are blue indicating that the virus is pure, stable, and expressing the foreign gene. Example 19G

Shipping Fever Vaccine

Shipping fever or bovine respiratory disease (BRD) complex is manifested as the result of a combination of infectious diseases of cattle and additional stress related factors (52) . Respiratory virus infections augmented by pathophysiological effects of stress, alter the susceptibility of cattle to Pasteurella organisms by a number of mechanisms. Control of the viral infections that initiate BRD is essential to preventing the disease syndrome (53) .

The major infectious disease pathogens that contribute to BRD include but are not limited to infectious bovine rhinotracheitis virus (IBRV) , parainfluenza virus type 3 (PI-3) , bovine respiratory syncytial virus (BRSV) , and Pasteurella haemolytica (53) . Recombinant swinepox virus expressing protective antigens to organisms causing BRD is useful as a vaccine. S-SPV-020, S-SPV- 029, S-SPV-030, S-SPV-032, and S-SPV-028 are useful components of such a vaccine.

Example 20

Recombinant swinepox viruses S-SPV-031 and S-SPV-035 are useful as a vaccine against human disease. S-SPV- 031 expresses the core antigen of hepatitis B virus. S- SPV-031 is useful against hepatitis B infection in humans. S-SPV-035 expresses the cytokine, interleukin- 2 , and is useful as an immune modulator to enhance an immune response in humans. When S-SPV-031 and S-SPV-035 are combined, a superior vaccine against hepatitis B is produced. Example 20A

S - SPV- 031 :

S-SPV-031 is a swinepox virus that expresses at least two foreign genes. The gene for E. coli _/S-galactosidase (lacZ) and the gene for Hepatitis B Core antigen were inserted into the SPV 617-48.1 ORF (a unique NotI restriction site has replaced a unique AccI restriction site) . The lacZ gene is under the control of the synthetic late promoter (LPl) , and the Hepatitis B Core antigen gene is under the control of the synthetic early/late promoter (EP1LP2) .

S-SPV-031 was derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing the homology vector 727-67.18 (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 _/S-galactosidase (BLUOGAL AND CPRG ASSAYS) . The final result of red plaque purification was the recombinant virus designated S-SPV-031. This virus was assayed for _/S-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 foreign gene.

S-SPV-031 was assayed for expression of Hepatitis B Core antigen-specific antigens using the BLACK PLAQUE SCREEN FOR FOREIGN GENE EXPRESSION IN RECOMBINANT SPV. Rabbit antisera to Hepatitis B Core antigen was shown to react specifically with S-SPV-031 plaques and not with S-SPV-001 negative control plaques. All S-SPV-031 observed plaques reacted with the antiserum indicating that the virus was stably expressing the Hepatitis B Core antigen gene. The assays described here were carried out in ESK-4 cells, indicating that ESK-4 cells would be a suitable substrate for the production of SPV recombinant vaccines.

To confirm the expression of the Hepatitis B Core antigen gene product, cells were infected with SPV-031 and samples of infected cell lysates were subjected to SDS-polyacryla ide gel electrophoresis. The gel was blotted and analyzed using the WESTERN BLOTTING PROCEDURE. Rabbit antisera to Hepatitis B Core antigen was used to detect expression of Hepatitis B specific proteins. The lysate from S-SPV-031 infected cells exhibited a band at 21 kd which is the expected size of the Hepatitis B Core antigen.

Example 2OB

S-SPV-035:

S-SPV-035 is a swinepox virus that expresses at least two foreign genes. The gene for E. coli _/β-galactosidase (lacZ) and the gene for human IL-2 were inserted into the SPV 617-48.1 ORF (a unique NotI restriction site has replaced a unique AccI restriction site) . The lacZ gene is under the control of the synthetic late promoter (LPl) , and the human IL-2 gene is under the control of the synthetic late/early promoter (LP2EP2) .

S-SPV-035 was derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing the homology vector 741-84.14 (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 _/8-galactosidase (BLUOGAL AND CPRG ASSAYS) . The final result of red plaque purification was the recombinant virus designated S-SPV-035. This virus was assayed for iβ-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 foreign gene.

Example 21

Human vaccines using recombinant swinepox virus as a vector

Recombinant swinepox virus is useful as a vaccine against human diseases. For example, human influenza virus is a rapidly evolving virus whose neutralizing viral epitopes rapidly change. A useful recombinant swinepox vaccine is one in which the influenza virus neutralizing epitopes are quickly adapted by recombinant DNA techniques to protect against new strains of influenza virus. Human influenza virus hemagglutinin (HN) and neuraminidase (NA) genes are cloned into the swinepox virus as described in CLONING OF EQUINE INFLUENZA VIRUS HEMAGGLUTININ AND NEURAMINIDASE GENES (See Materials and Methods and Example 17) .

Recombinant swinepox virus is useful as a vaccine against other human diseases when foreign antigens from the following diseases or disease organisms are expressed in the swinepox virus vector: hepatitis B virus surface and core antigens, hepatitis C virus, human immunodeficiency virus, human herpesviruses, herpes simplex virus-1, herpes simplex virus-2, human cytomegalovirus, Epstein-Barr virus, Varicella-Zoster virus, human herpesvirus-6, human herpesvirus-7, human influenza, measles virus, hantaan virus, pneumonia virus, rhinovirs, poliovirus, human respiratory syncytial virus, retrovirus, human T-cell leukemia virus, rabies virus, mumps virus, malaria ( Plasmodium falciparum) , Bordetelia pertussis, Diptheria, Rickettsia prowazekii , Borrelia bergdorferi , Tetanus toxoid, malignant tumor antigens. Furthermore, S-SPV-035 (Example 20) , when combined with swinepox virus interleukin-2 is useful in enhancing immune response in humans. Additional cytokines, including but not limited to, interleukin-2, interleukin-6 , interleukin-12 , interferons, granulocyte-macrophage colony stimulating factors, interleukin receptors from human and other animals when vectored into a non-essential site in the swinepox viral genome, and subsequently expressed, have immune stimulating effects.

Recombinant swinepox virus express foreign genes in a human cell line. S-SPV-003 (EP1LP2 promoter expressing the lacZ gene) expressed the lacZ gene in THP human monocyte cell lines by measuring _/8-galactosidase activity. Cytopathic effect of swinepox virus was observed on the THP human monocyte cells, indicating that recombinant swinepox virus can express foreign genes in a human cell line, but will not productively infect or replicated in the human cell line. Swinepox virus was demonstrated to replicate well in ESK-4 cells (embryonic swine kidney) indicating that ESK-4 cells would be a suitable substrate for the production of SPV recombinant vaccines.

Example 22

Avian vaccines using recombinant swinepox virus as a vector.

Example 22A

S-SPV-026

S-SPV-026 is a swinepox virus that expresses at least two foreign genes. The gene for E. coli /β-galactosidase (lacZ) and the gene for infectious bursal disease virus (IBDV) polyprotein were inserted into the SPV 617-48.1 ORF (a unique NotI restriction site has replaced a unique AccI restriction site) . The lacZ gene is under the control of the synthetic late promoter (LPl) , and the IBDV polyprotein gene is under the control of the synthetic early/late promoter (EP1LP2) .

S-SPV-026 was derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing the homology vector 689-50.4 (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-026. This virus was assayed for /S-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 indication that the virus was pure, stable, and expressing the foreign gene.

S-SPV-026 was assayed for expression of IBDV polyprotein-specific antigens using the BLACK PLAQUE

SCREEN FOR FOREIGN GENE EXPRESSION IN RECOMBINANT SPV.

Rat antisera to IBDV polyprotein were shown to react specifically with S-SPV-026 plaques and not with S-SPV-

001 negative control plaques. All S-SPV-026 observed plaques reacted with the antiserum indicating that the virus was stably expressing the IBDV polyprotein gene.

The assays described here were carried out in ESK-4 cells, indicating that ESK-4 cells would be a suitable substrate for the production of SPV recombinant vaccines. To confirm the expression of the IBDV polyprotein gene product, cells were infected with SPV-026 and samples of infected cell lysates were subjected to SDS- polyacrylamide gel electrophoresis. The gel was blotted and analyzed using the WESTERN BLOTTING PROCEDURE. Rat antisera to IBDV proteins VP2, VP3, and VP4 and monoclonal antibody R63 to IBDV VP2 were used to detect expression of IBDV proteins. The lysate from S-SPV-026 infected cells exhibited bands at 32 to 40 kd which is the expected size of the IBDV proteins.

Example 22B

S-SPV-027

S-SPV-027 is a swinepox virus that expresses at least two foreign genes. The gene for E. coli _/β-galactosidase (lacZ) and the gene for infectious bursal disease virus (IBDV) VP2 (40kd) were inserted into the SPV 617-48.1 ORF (a unique NotI restriction site has replaced a unique AccI restriction site) . The lacZ gene is under the control of the synthetic late promoter (LPl) , and the IBDV VP2 gene is under the control of the synthetic early/late promoter (EP1LP2) .

S-SPV-027 was derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing the homology vector 689-50.7 (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 /3-galactosidase (BLUOGAL AND CPRG ASSAYS) . The final result of red plaque purification was the recombinant virus designated S-SPV-027. This virus was assayed for _/S-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 foreign gene.

S-SPV-027 was assayed for expression of IBDV VP2- specific antigens using the BLACK PLAQUE SCREEN FOR FOREIGN GENE EXPRESSION IN RECOMBINANT SPV. Rat antisera to IBDV protein was shown to react specifically with S-SPV-027 plaques and not with S-SPV- 001 negative control plaques. All S-SPV-027 observed plaques reacted with the antiserum indicating that the virus was stably expressing the IBDV VP2 gene. The assays described here were carried out in ESK-4 cells, indicating that ESK-4 cells would be a suitable substrate for the production of SPV recombinant vaccines .

To confirm the expression of the IBDV VP2 gene product, cells were infected with S-SPV-027 and samples of infected cell lysates were subjected to SDS- polyacrylamide gel electrophoresis. The gel was blotted and analyzed using the WESTERN BLOTTING PROCEDURE. Rat antisera to IBDV protein and monoclonal antibody R63 to IBDV VP2 were used to detect expression of IBDV VP2 protein. The lysate from S-SPV-027 infected cells exhibited a band at 40 kd which is the expected size of the IBDV VP2 protein.

S-SPV-026 and S-SPV-027 are useful as vaccines against infectious bursal disease in chickens and also as expression vectors for IBDV proteins. Recombinant swinepox virus is useful as a vaccine against other avian disease when foreign antigens from the following diseases or disease organisms are expressed in the swinepox virus vector: Marek's disease virus, infectious laryngotracheitis virus, Newcastle disease virus, infectious bronchitis virus, and chicken anemia virus, Chick anemia virus, Avian encephalomyelitis virus, Avian reovirus, Avian paramyxoviruses, Avian influenza virus, Avian adenovirus, Fowl pox virus, Avian coronavirus, Avian rotavirus, Salmonella spp E coli , Pasteurella spp, Haemophilus spp, Chlamydia spp, Mycoplasma spp, Campylobacter spp, Bordetella spp, Poultry nematodes, cestodes, trematodes, Poultry mites/lice, Poultry protozoa {Eimeria spp, Histomonas spp, Trichomonas spp) .

Example 23

SPV-036:

S-SPV-036 is a swinepox virus that expresses at one foreign gene. The gene for E. coli _/8-galactosidase

(lacZ) was inserted into the SPV 617-48.1 ORF (a unique

NotI restriction site has replaced a unique AccI restriction site) . The lacZ gene is under the control of the human cytomegalovirus immediate early (HCMV IE) promoter.

S-SPV-036 was derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing the homology vector 741-80.3 (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 _/8-galactosidase (BLUOGAL AND CPRG ASSAYS) . The final result of red plaque purification was the recombinant virus designated S-SPV-036. This virus is assayed for _/S-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 are blue indicating that the virus is pure, stable, and expressing the foreign gene. The expression of lacZ from the HCMV IE promoter provides a strong promoter for expression of foreign genes in swinepox. S-SPV-036 is a novel and unexpected demonstration of a herpesvirus promoter driving expression of a foreign gene in a poxvirus . S-SPV-036 is useful in formulating human vaccines, and recombinant swinepox virus is useful for the expression of neutralizing antigens from human pathogens. Recombinant swinepox virus expressed foreign genes in a human cell line as demonstrated by S-SPV-003 (EP1LP2) promoter expressing the lacZ gene) expressed β- galactosidase in THP human monocyte cell lines. Cytopathic effects of swinepox virus on the THP human monocyte cells were not observed, indicating that recombinant swinepox virus can express foreign genes in a human cell line, but will not productively infect or replicated in the human cell line

Example 24

Homology Vector 738-94.4

Homology Vector 738-94.4 is a swinepox virus vector that expresses one foreign gene. The gene for E. coli -S-galactosidase (lacZ) was inserted into the the OIL open reading frame (SEQ ID NO: 115) . The lacZ gene is under the control of the OIL promoter. The homology vector 738-94.4 contains a deletion of SPV DNA from nucleotides 1679 to 2452 (SEQ ID NO: 189; Figure 17) which deletes part of the OIL ORF.

The upstream SPV sequences were synthesized by polymerase chain reaction using DNA primers 5'- GAAGCATGCCCGTTCTTATCAATAGTTTAGTCGAAAATA-3' (SEQ ID NO: 185) and 5' -CATAAGATCTGGCATTGTGTTATTATACTAACAAAAATAAG- 3' (SEQ ID NO: 186) to produce an 855 base pair fragment with Bglll and SphI ends. The OIL promoter is present on this fragment. The downstream SPV sequences were synthesized by polymerase chain reaction using DNA primers 5' -CCGTAGTCGACΑAAGATCGACTTATTAATATGTATGGGATT-3' ( S E Q I D N O : 1 8 7 ) a n d 5 ' - GCCTGAAGCTTCTAGTACAGTATTTACGACTTTTGAAAT-3' (SEQ ID NO: 188) to produce an 1113 base pair fragment with Sail and Hindlll ends. A recombinant swinepox virus was derived utilizing homology vector 738-94.4 and S-SPV- 001 (Kasza strain) in the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT SPV. The transfection stock was screened by the SCREEN FOR RECOMBINANT SPV EXPRESSING _/8-galactosidase (BLUOGAL AND CPRG ASSAYS) . The final result of red plaque purification is the recombinant virus. This virus is assayed for 3-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 are blue indicating that the virus is pure, stable, and expressing the foreign gene. Recombinant swinepox viruses derived from homology vector 738-94.4 are utilized as an expression vector to express foreign antigens and as a vaccine to raise a protective immune response in animals to foreign genes expressed by the recombinant swinepox virus. Other promoters in addition to the OIL promoter are inserted into the deleted region including LPl, EP1LP2, LP2EP2, HCMV immediate early, and one or more foreign genes are expressed from these promoters .

Example 24B

Homology Vector 752-22.1 is a swinepox virus vector that is utilized to express two foreign genes. The gene for E. coli _/S-galactosidase (lacZ) was inserted into the the OIL open reading frame (SEQ ID NO: 115) . The lacZ gene is under the control of the OIL promoter. A second foreign gene is expressed from the LP2EP2 promoter inserted into an EcoRI or BamHI site following the LP2EP2 promoter sequence. The homology vector 752- 22.1 contains a deletion of SPV DNA from nucleotides 1679 to 2452 (SEQ ID NO: 189; Figure 17) which deletes part of the OIL ORF. The homology vector 752-22.1 was derived from homology vector 738-94.4 by insertion of the LP2EP2 promoter fragment (see Materials and Methods) . The homology vector 752-22.1 is further improved by placing the lacZ gene under the control of the synthetic LPl promoter. The LPl promoter results in higher levels of lacZ expression compared to the SPV OIL promoter

Example 25

S-SPV-041:

S-SPV-041 is a swinepox virus that expresses at least two foreign genes. The gene for E. coli _/3-galactosidase (lacZ) and the gene for equine herpesvirus type 1 glycoprotein B (gB) were inserted into the 738-94.4 ORF (a 773 base pair deletion of the SPV OIL ORF; Deletion of nucleotides 1679 to 2452, SEQ ID NO: 189) . The lacZ gene is under the control of the swinepox OIL promoter, and the EHV-1 gB gene is under the control of the synthetic late/early promoter (LP2EP2) .

S-SPV-041 was derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing the homology vector 752-29.33 (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 _/S-galactosidase (BLUOGAL AND CPRG ASSAYS) . The final result of red plaque purification was the recombinant virus designated S-SPV-041. This virus was assayed for 3-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 foreign gene.

S-SPV-041 is useful as a vaccine in horses against EHV- 1 infection and is useful for expression of EHV-1 glycoprotein B.

S-SPV-045:

S-SPV-045 is a swinepox virus that expresses at least two foreign genes. The gene for E. coli _/S-galactosidase (lacZ) and the gene for infectious bovine rhinotracheitis virus glycoprotein E (gE) were inserted into the 738-94.4 ORF (a 773 base pair deletion of the SPV OIL ORF; Deletion of nucleotides 1679 to 2452, SEQ ID NO: 189) . The lacZ gene is under the control of the swinepox OIL promoter, and the IBRV gE gene is under the control of the synthetic late/early promoter (LP2EP2) .

S-SPV-045 was derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing the homology vector 746-94.1 (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 _/S-galactosidase (BLUOGAL AND CPRG ASSAYS) . The final result of red plaque purification was the recombinant virus designated S-SPV-045. 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 foreign gene.

S-SPV-045 is useful for expression of IBRV glycoprotein E.

S-SPV-049:

S-SPV-049 is a swinepox virus that expresses at least two foreign genes. The gene for E. coli _/S-galactosidase

(lacZ) and the gene for bovine viral diarrhea virus glycoprotein 48 (gp48) were inserted into the 738-94.4

ORF (a 773 base pair deletion of the SPV OIL ORF;

Deletion of nucleotides 1679 to 2452, SEQ ID NO: 189) . The lacZ gene is under the control of the swinepox OIL promoter, and the BVDV gp48 gene is under the control of the synthetic late/early promoter (LP2EP2) .

S-SPV-049 was derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing the homology vector 771-55.11 (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 /3-galactosidase (BLUOGAL AND CPRG ASSAYS) . The final result of red plaque purification was the recombinant virus designated S-SPV-049. 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 foreign gene.

S-SPV-049 is useful as a vaccine in cattle against BVDV infection and is useful for expression of BVDV glycoprotein 48. S - SPV- 050 :

S-SPV-050 is a swinepox virus that expresses at least two foreign genes. The gene for E. coli S-galactosidase

(lacZ) and the gene for the bovine viral diarrhea virus glycoprotein 53 (gp53) were inserted into the 738-94.4

ORF (a 773 base pair deletion of the SPV OIL ORF;

Deletion of nucleotides 1679 to 2452, SEQ ID NO: 189) . The lacZ gene is under the control of the swinepox OIL promoter, and the IBRV gE gene is under the control of the synthetic late/early promoter (LP2EP2) .

S-SPV-050 was derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing the homology vector 767-67.3 (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-050. This virus was assayed for _/S-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 foreign gene.

S-SPV-050 is useful as a vaccine in cattle against BVDV infection and is useful for expression of BVDV glycoprotein 53.

Example 26

Recombinant swinepox virus, S-SPV-042 or S-SPV-043, expressing chicken interferon (cIFN) or chicken myelomonocytic growth factor (cMGF) , respectively, are useful to enhance the immune response when added to vaccines against diseases of poultry. Chicken myelomonocytic growth factor (cMGF) is homologous to mammalian interleukin-6 protein, and chicken interferon

(cIFN) is homologous to mammalian interferon. When used in combination with vaccines against specific avian diseases, S-SPV-042 and S-SPV-043 provide enhanced mucosal, humoral, or cell mediated immunity against avian disease-causing viruses including, but not limited to, Marek's disease virus, Newcastle disease virus, infectious laryngotracheitis virus, infectious bronchitis virus, infectious bursal disease virus.

Example 26A

S-SPV-042:

S-SPV-042 is a swinepox virus that expresses at least two foreign genes. The gene for E. coli _/S-galactosidase

(lacZ) and the gene for chicken interferon (cIFN) were inserted into the SPV 617-48.1 ORF (a unique NotI restriction site has replaced a unique AccI restriction site) . The lacZ gene is under the control of the synthetic late promoter (LPl) , and the cIFN gene is under the control of the synthetic late/early promoter

(LP2EP2) .

S-SPV-042 was derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing the homology vector 751-07.Al (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 _/S-galactosidase (BLUOGAL AND CPRG ASSAYS) . The final result of red plaque purification was the recombinant virus designated S-SPV-042. This virus was assayed for _/8-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 foreign gene.

S-SPV-042 has interferon activity in cell culture.

Addition of S-SPV-042 conditioned media to chicken embryo fibroblast (CEF) cell culture inhibits infection of the CEF cells by vesicular stomatitis virus or by herpesvirus of turkeys. S-SPV-042 is useful to enhance the immune response when added to vaccines against diseases of poultry.

Example 26B

S-SPV-043:

S-SPV-043 is a swinepox virus that expresses at least two foreign genes. The gene for E. coli _/8-galactosidase (lacZ) and the gene for chicken myelomonocytic growth factor (cMGF) were inserted into the SPV 617-48.1 ORF (a unique NotI restriction site has replaced a unique AccI restriction site) . The lacZ gene is under the control of the synthetic late promoter (LPl) , and the cMGF gene is under the control of the synthetic late/early promoter (LP2EP2) .

S-SPV-043 was derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing the homology vector 751-56.Al (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 _/8-galactosidase (BLUOGAL AND CPRG ASSAYS) . The final result of red plaque purification was the recombinant virus designated S-SPV-043. This virus was assayed for /8-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 foreign gene.

S-SPV-043 is useful to enhance the immune response when added to vaccines against diseases of poultry.

Example 27

Insertion into a non-essential site in the 2.0 kb Hindlll to Bglll region of the swinepox virus Hindlll M fragment .

A 2.0 kb Hindlll to Bglll region of the swinepox virus Hindlll M fragment is useful for the insertion of foreign DNA into SPV. The foreign DNA is inserted into a unique Bglll restriction site in the region (Figure 17; Nucleotide 540 of SEQ ID NOs: 195) . A plasmid containing a foreign DNA insert is used according to the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT SPV to generate an 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 Bglll restriction site in the 2.0 kb Hindlll to Bglll region of the swinepox virus Hindlll M fragment is located within the coding region of the SPV I4L open reading frame. The I4L ORF has sequence similarity to the vaccinia virus and smallpox virus ribonucleotide reductase (large subunit) gene

(56-58) . The ribonucleotide reductase (large subunit) gene is non-essential for DNA replication of vaccinia virus and is an appropriate insertion site in swinepox virus.

Example 28

S-SPV-047

S-SPV-047 is a swinepox virus that expresses two foreign genes. The gene for E. coli /3-galactosidase

(lacZ) and the gene for pseudorabies virus gB (gll) were inserted into a unique Hindlll site (Hindlll linker inserted into the Bglll restriction endonuclease site within the 2.0 kb Bglll to Hindlll subfragment of the Hindlll M fragment.) The Bglll insertion site is within the SPV I4L open reading frame which has significant homology to the vaccinia virus ribonucleoside-diphosphate reductase gene. The lacZ gene is under the control of the synthetic late promoter (LPl) , and the PRV gB (gll) gene is under the control of the synthetic late/early promoter (LP2EP2) .

S-SPV-047 was derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing the homology vector 779-94.31 (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 /S-galactosidase (BLUOGAL AND CPRG ASSAYS) . The final result of red plaque purification was the recombinant virus designated S-SPV-047. This virus was assayed for /S-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 foreign gene. S-SPV-047 was assayed for expression of PRV specific antigens using the BLACK PLAQUE SCREEN FOR FOREIGN GENE EXPRESSION IN RECOMBINANT SPV. Polyclonal swine anti- PRV serum was shown to react specifically with S-SPV- 047 plaques and not with S-SPV-001 negative control plaques. All S-SPV-047 observed plaques reacted with the swine anti-PRV serum indicating that the virus was stably expressing the PRV foreign gene. The assays described here were carried out in ESK-4 cells, indicating that ESK-4 cells would be a suitable substrate for the production of SPV recombinant vaccines .

To confirm the expression of the PRV gB gene product, cells were infected with S-SPV-047 and samples of infected cell lysates and culture supernatants were subjected to SDS polyacrylamide gel electrophoresis. The gel was blotted and analyzed using the WESTERN BLOTTING PROCEDURE. A polyclonal swine anti-PRV serum was used to detect expression of PRV specific proteins. The cell lysate and supernatants from S-SPV-047 infected cells exhibited bands corresponding to 120 kD, 67 kD and 58 kD, which are the expected size of the PRV glycoprotein B.

SPV recombinant-expressed PRV gB has been shown to elicit a significant immune response in swine (37, 38; See example 8) . Furthermore, PRV gB is expressed in recombinant SPV, significant protection from challenge with virulent PRV is obtained. (See Examples 6 and 8) Therefore S-SPV-047 is valuable as a vaccine to protect swine against PRV disease. Since the PRV vaccines described here do not express PRV gX or gl, 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- 052

S-SPV-052 is a swinepox virus that expresses three foreign genes. The gene for E. coli _/β-galactosidase

(lacZ) and the gene for pseudorabies virus gB (gll) were inserted into the unique Hindlll restriction site

(Hindlll linkers inserted into a unique Ndel site in the SPV OIL open reading frame; An approximately 545 base pair Ndel to Ndel subfragment (Nucleotides 1560 to 2104; SEQ ID NO. ) of the SPV Hindlll M fragment has been deleted) . The gene for PRV gD (g50) was inserted into the unique PstI restriction site (PstI linkers inserted into a unique AccI site in the SPV OIL open reading frame) . The lacZ gene is under the control of the synthetic late promoter (LPl) , the PRV gB (gll) gene is under the control of the synthetic late/early promoter (LP2EP2) , and the PRV gD (g50) gene is under the control of the synthetic early/late promoter (EP1LP2) .

S-SPV-052 was derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing the homology vector 789-41.7 (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 _/S-galactosidase (BLUOGAL AND CPRG ASSAYS) . The final result of red plaque purification was the recombinant virus designated S-SPV 052. This virus was assayed for _/S-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 foreign gene. S-SPV-052 was assayed for expression of PRV specific antigens using the BLACK PLAQUE SCREEN FOR FOREIGN GENE EXPRESSION IN RECOMBINANT SPV. Polyclonal swine anti- PRV serum was shown to react specifically with S-SPV- 052 plaques and not with S-SPV-001 negative control plaques. All S-SPV-052 observed plaques reacted with the swine anti-PRV serum indicating that the virus was stably expressing the PRV foreign gene. The assays described here were carried out in ESK-4 cells, indicating that ESK-4 cells would be a suitable substrate for the production of SPV recombinant vaccines .

To confirm the expression of the PRV gB and gD gene products, cells were infected with S-SPV-052 and samples of infected cell lysates and culture supernatants were subjected to SDS polyacrylamide gel electrophoresis. The gel was blotted and analyzed using the WESTERN BLOTTING PROCEDURE. A polyclonal swine anti-PRV serum was used to detect expression of

PRV specific proteins. The cell lysate and supernatants from S-SPV-052 infected cells exhibited bands corresponding to 120 kD, 67 kD and 58 kD, which are the expected size of the PRV glycoprotein B; and a 48 kD which is the expected size of the PRV glycoprotein D.

SPV recombinant-expressed PRV gB and gD has been shown to elicit a significant immune response in swine (37, 38; See example 8) . Furthermore, PRV gB and gD are expressed in recombinant SPV, significant protection from challenge with virulent PRV is obtained. (See Examples 6 and 8) Therefore S-SPV-052 is valuable as a vaccine to protect swine against PRV disease. Since the PRV vaccines described here do not express PRV gX or gl, 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-053

S-SPV-053 is a swinepox virus that expresses three foreign genes. The gene for E. coli /S-galactosidase

(lacZ) and the gene for pseudorabies virus gB (gll) were inserted into the unique Hindlll restriction site (Hindlll linkers inserted into a unique Ndel site in the SPV OIL open reading frame; An approximately 545 base pair Ndel to Ndel subfragment (Nucleotides 1560 to 2104; SEQ ID NO. ) of the SPV Hindlll M fragment has been deleted) . The gene for PRV gC (gill) was inserted into the unique PstI restriction site (PstI linkers inserted into a unique AccI site in the SPV OIL open reading frame) . The lacZ gene is under the control of the synthetic late promoter (LPl) , the PRV gB (gll) gene is under the control of the synthetic late/early promoter (LP2EP2) , and the PRV gC (gill) gene is under the control of the synthetic early/late promoter (EP1LP2) .

S-SPV-053 was derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing the homology vector 789-41.27 (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 _/S-galactosidase (BLUOGAL AND CPRG ASSAYS) . The final result of red plaque purification was the recombinant virus designated S-SPV 053. This virus was assayed for _/S-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 foreign gene.

S-SPV-053 was assayed for expression of PRV specific antigens using the BLACK PLAQUE SCREEN FOR FOREIGN GENE EXPRESSION IN RECOMBINANT SPV. Polyclonal swine anti- PRV serum was shown to react specifically with S-SPV- 053 plaques and not with S-SPV-001 negative control plaques. All S-SPV-053 observed plaques reacted with the swine anti-PRV serum indicating that the virus was stably expressing the PRV foreign gene. The assays described here were carried out in ESK-4 cells, indicating that ESK-4 cells would be a suitable substrate for the production of SPV recombinant vaccines.

To confirm the expression of the PRV gB and gC gene products, cells were infected with S-SPV-053 and samples of infected cell lysates and culture supernatants were subjected to SDS polyacrylamide gel electrophoresis. The gel was blotted and analyzed using the WESTERN BLOTTING PROCEDURE. A polyclonal swine anti-PRV serum was used to detect expression of PRV specific proteins. The cell lysate and supernatants from S-SPV-053 infected cells exhibited bands corresponding to 120 kD, 67 kD and 58 kD, which are the expected size of the PRV glycoprotein B; and a 92 kD which is the expected size of the PRV glycoprotein C.

SPV recombinant-expressed PRV gB and gC has been shown to elicit a significant immune response in swine (37, 38; See example 8) . Furthermore, PRV gB and gC are expressed in recombinant SPV, significant protection from challenge with virulent PRV is obtained. (See Examples 6 and 8) Therefore S-SPV-053 is valuable as a vaccine to protect swine against PRV disease. Since the PRV vaccines described here do not express PRV gX or gl, 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-054

S-SPV-054 is a swinepox virus that expresses three foreign genes. The gene for E. coli _/S-galactosidase

(lacZ) and the gene for pseudorabies virus gC (gill) were inserted into the unique Hindlll restriction site

(Hindlll linkers inserted into a unique Ndel site in the SPV OIL open reading frame; An approximately 545 base pair Ndel to Ndel subfragment (Nucleotides 1560 to 2104; SEQ ID NO. ) of the SPV Hindlll M fragment has been deleted) . The gene for PRV gD (g50) was inserted into the unique PstI restriction site (PstI linkers inserted into a unique AccI site in the SPV OIL open reading frame) . The lacZ gene is under the control of the synthetic late promoter (LPl) , the PRV gC (gill) gene is under the control of the synthetic early/late promoter (EP1LP2) , and the PRV gD (g50) gene is under the control of the synthetic early/late promoter (EP1LP2) .

S-SPV-054 was derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing the homology vector 789-41.47 (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 _/S-galactosidase (BLUOGAL AND CPRG ASSAYS) . The final result of red plaque purification was the recombinant virus designated S-SPV 054. This virus was assayed for /3-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 foreign gene.

S-SPV-054 was assayed for expression of PRV specific antigens using the BLACK PLAQUE SCREEN FOR FOREIGN GENE EXPRESSION IN RECOMBINANT SPV. Polyclonal swine anti- PRV serum was shown to react specifically with S-SPV- 054 plaques and not with S-SPV-001 negative control plaques. All S-SPV-054 observed plaques reacted with the swine anti-PRV serum indicating that the virus was stably expressing the PRV foreign gene. The assays described here were carried out in ESK-4 cells, indicating that ESK-4 cells would be a suitable substrate for the production of SPV recombinant vaccines.

To confirm the expression of the PRV gC and gD gene products, cells were infected with S-SPV-054 and samples of infected cell lysates and culture supernatants were subjected to SDS polyacrylamide gel electrophoresis. The gel was blotted and analyzed using the WESTERN BLOTTING PROCEDURE. A polyclonal swine anti-PRV serum was used to detect expression of

PRV specific proteins. The cell lysate and supernatants from S-SPV-054 infected cells exhibited a band corresponding to 92 kD which is the expected size of the PRV glycoprotein C and a 48 kD which is the expected size of the PRV glycoprotein D.

SPV recombinant-expressed PRV gC and gD has been shown to elicit a significant immune response in swine (37, 38; See example 8) . Furthermore, PRV gC and gD are expressed in recombinant SPV, significant protection from challenge with virulent PRV is obtained. (See Examples 6 and 8) Therefore S-SPV-054 is valuable as a vaccine to protect swine against PRV disease. Since the PRV vaccines described here do not express PRV gX or gl, 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-055

S-SPV-055 is a swinepox virus that expresses four foreign genes. The gene for E. coli /S-galactosidase

(lacZ) and the gene for pseudorabies virus gB (gll) were inserted into the unique Hindlll restriction site

(Hindlll linkers inserted into a unique Ndel site in the SPV OIL open reading frame; An approximately 545 base pair Ndel to Ndel subfragment (Nucleotides 1560 to 2104; SEQ ID NO. ) of the SPV Hindlll M fragment has been deleted) . The gene for PRV gD (g50) and PRV gC (gill) were inserted into the unique PstI restriction site (PstI linkers inserted into a unique AccI site in the SPV OIL open reading frame) . The lacZ gene is under the control of the synthetic late promoter (LPl) , the PRV gB (gll) gene is under the control of the synthetic late/early promoter (LP2EP2) , the PRV gD (g50) gene is under the control of the synthetic late/early promoter (LP2EP2) , and the PRV gC (gill) gene is under the control of the synthetic early/late promoter (EP1LP2) .

S-SPV-055 was derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing the homology vector 789-41.73 (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 _/S-galactosidase (BLUOGAL AND CPRG ASSAYS) . The final result of red plaque purification was the recombinant virus designated S-SPV 055. This virus was assayed for /S-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 foreign gene.

S-SPV-055 was assayed for expression of PRV specific antigens using the BLACK PLAQUE SCREEN FOR FOREIGN GENE EXPRESSION IN RECOMBINANT SPV. Polyclonal swine anti- PRV serum was shown to react specifically with S-SPV- 055 plaques and not with S-SPV-001 negative control plaques. All S-SPV-055 observed plaques reacted with the swine anti-PRV serum indicating that the virus was stably expressing the PRV foreign gene. The assays described here were carried out in ESK-4 cells, indicating that ESK-4 cells would be a suitable substrate for the production of SPV recombinant vaccines.

To confirm the expression of the PRV gB, gC and gD gene products, cells were infected with S-SPV-055 and samples of infected cell lysates and culture supernatants were subjected to SDS polyacrylamide gel electrophoresis. The gel was blotted and analyzed using the WESTERN BLOTTING PROCEDURE. A polyclonal swine anti-PRV serum was used to detect expression of PRV specific proteins. The cell lysate and supernatants from S-SPV-055 infected cells exhibited a bands corresponding to 120 kD, 67 kD, and 58 kD which is the expected size of the PRV glycoprotein B; a 92 kD which is the expected size of the PRV glycoprotein C; and a 48 kD which is the expected size of the PRV glycoprotein D

SPV recombinant-expressed PRV gB, gC and gD has been shown to elicit a significant immune response in swine (37, 38; See example 8) . Furthermore, PRV gB, gC and gD are expressed in recombinant SPV, significant protection from challenge with virulent PRV is obtained. (See Examples 6 and 8) Therefore S-SPV-055 is valuable as a vaccine to protect swine against PRV disease. Since the PRV vaccines described here do not express PRV gX or gl, 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.

Example 29

SPV-059

S-SPV-059 is a swinepox virus that expresses one foreign gene. The gene for E. coli _/S-glucuronidase (uidA) was inserted into the unique EcoRI restriction site in the SPV B18R open reading frame within the SPV Hindlll K genomic fragment. The uidA gene is under the control of the synthetic late/early promoter (LP2EP2) . Partial sequence from a 3.2 kb region of the SPV 6.5 kb Hindlll K fragment (SEQ ID NO. ) indicates three potential open reading frames. The SPV B18R ORF shows sequence homology to the vaccinia virus B18R gene, 77.2K protein from rabbit fibroma virus, vaccinia virus C19L/B25R ORF and an ankyrin repeat region from a human brain variant. The B18R gene codes for a soluble interferon receptor with high affinity and broad specificty. The SPV B4R open reading frame shows sequence homology to the T5 protein of rabbit fibroma virus .

S-SPV-059 was derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing the homology vector

796-50.31 and virus S-SPV-001 in the HOMOLOGOUS

RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT SPV. Homology vector 796-50.31 was generated by insertion of a blunt ended NotI fragment containing the LP2EP2 promoter uidA cassette from plasmid 551-47.23 (see Materials and Methods) into a unique EcoRI site (blunt ended) in the SPV 6.5 kb Hindlll K fragment, (Figure 29B) . The transfection stock was screened by the SCREEN FOR RECOMBINANT HERPESVIRUS EXPRESSING ENZYMATIC MARKER GENES. The final result of blue plaque purification was the recombinant virus designated S- SPV-059. This virus was assayed for _/8-glucuronidase 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, plaques observed were blue indicating that the virus was pure, stable, and expressing the foreign gene.

S-SPV-059 has been purified and expresses the foreign gene, E. coli uidA, indicating that the EcoRI site within the 6.5 kb Hindlll K fragment is a stable insertion site for foreign genes . Recombinant swinepox virus utilizing this insertion site is useful for expression of foreign antigen genes, as a vaccine against disease or as an expression vector to raise antibodies to the expressed foreign gene.

SPV-060

S-SPV-060 is a swinepox virus that expresses one foreign gene. The gene for E. coli β-glucuronidase

(uidA) was inserted into the unique EcoRV restriction site within the SPV Hindlll N genomic fragment. The uidA gene is under the control of the synthetic late/early promoter (LP2EP2) . Partial sequence of the SPV 3.2 kb Hindlll N fragment (SEQ ID NO. ) indicates two potential open reading frames. The SPV

I7L ORF shows sequence homology to protein 17 of vaccinia virus. The SPV I4L open reading frame shows sequence homology to the ribonucleoside diphosphate reductase gene of vaccinia virus. Two potential open reading frames I5L and I6L, between I4L ORF and I7L ORF are of unknown function.

S-SPV-060 was derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing the homology vector 796-71.31 and virus S-SPV-001 in the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT

SPV. Homology vector 796-71.31 was generated by insertion of a blunt ended ΝotI fragment containing the

LP2EP2 promoter uidA cassette from plasmid 551-47.23

(see Materials and Methods) into a unique EcoRV site in the SPV 3.2 kb Hindlll Ν fragment (Figure 29A) . The transfection stock was screened by the SCREEN FOR RECOMBINANT HERPESVIRUS EXPRESSING ENZYMATIC MARKER GENES. The final result of blue plaque purification is the recombinant virus designated S-SPV-060. This virus is assayed for _/β-glucuronidase 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, plaques observed are blue indicating that the virus is pure, stable, and expressing the foreign gene.

S-SPV-060 is purified and expresses the foreign gene, E. coli uidA, indicating that the EcoRI site within the 3.2 kb Hindlll N fragment is a stable insertion site for foreign genes. Recombinant swinepox virus utilizing this insertion site is useful for expression of foreign antigen genes, as a vaccine against disease or as an expression vector to raise antibodies to the expressed foreign gene.

S-SPV-061 S-SPV-061 is a swinepox virus that expresses one foreign gene. The gene for E. coli _/β-glucuronidase (uidA) was inserted into the unique SnaBI restriction site within the SPV Hindlll N genomic fragment. The uidA gene is under the control of the synthetic late/early promoter (LP2EP2) . Partial sequence of the SPV 3.2 kb Hindlll N fragment (SEQ ID NO. ) indicates two potential open reading frames. The SPV I7L ORF shows sequence homology to protein 17 of vaccinia virus. The SPV I4L open reading frame shows sequence homology to the ribonucleoside diphosphate reductase gene of vaccinia virus. Two potential open reading frames I5L and I6L, between I4L ORF and I7L ORF are of unknown function.

S-SPV-061 was derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing the homology vector 796-71.41 and virus S-SPV-001 in the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT SPV. Homology vector 796-71.41 was generated by insertion of a blunt ended NotI fragment containing the LP2EP2 promoter uidA cassette from plasmid 551-47.23 (see Materials and Methods) into a unique SnaBI site in the SPV 3.2 kb Hindlll N fragment. The transfection stock was screened by the SCREEN FOR RECOMBINANT HERPESVIRUS

EXPRESSING ENZYMATIC MARKER GENES. The final result of blue plaque purification is the recombinant virus designated S-SPV-061. This virus is assayed for β- glucuronidase 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, plaques observed are blue indicating that the virus is pure, stable, and expressing the foreign gene.

S-SPV-061 is purified and expresses the foreign gene, E. coli uidA, indicating that the SnaBI site within the 3.2 kb' Hindlll N fragment is a stable insertion site for foreign genes. Recombinant swinepox virus utilizing this insertion site is useful for expression of foreign antigen genes, as a vaccine against disease or as an expression vector to raise antibodies to the expressed foreign gene.

S-SPV-062

S-SPV-062 is a swinepox virus that expresses one foreign gene. The gene for E. coli _/S-glucuronidase (uidA) was inserted into the unique Bglll restriction site within the SPV Hindlll N genomic fragment (Figure 29A) . The uidA gene is under the control of the synthetic late/early promoter (LP2EP2) . Partial sequence of the SPV 3.2 kb Hindlll N fragment (SEQ ID NO. ) indicates two potential open reading frames. The SPV I7L ORF shows sequence homology to protein 17 of vaccinia virus. The SPV I4L open reading frame shows sequence homology to the ribonucleoside diphosphate reductase gene of vaccinia virus. Two potential open reading frames I5L and I6L, between I4L ORF and I7L ORF are of unknown function.

S-SPV-062 was derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing the homology vector 796-71.51 and virus S-SPV-001 in the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT SPV. Homology vector 796-71.51 was generated by insertion of a blunt ended NotI fragment containing the LP2EP2 promoter uidA cassette from plasmid 551-47.23 (see Materials and Methods) into a unique Bglll site in the SPV 3.2 kb Hindlll N fragment. The transfection stock was screened by the SCREEN FOR RECOMBINANT HERPESVIRUS EXPRESSING ENZYMATIC MARKER GENES. The final result of blue plaque purification is the recombinant virus designated S-SPV-062. This virus is assayed for β- glucuronidase 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, plaques observed are blue indicating that the virus is pure, stable, and expressing the foreign gene.

S-SPV-062 is purified and expresses the foreign gene, E. coli uidA, indicating that the Bglll site within the 3.2 kb Hindlll N fragment is a stable insertion site for foreign genes. Recombinant swinepox virus utilizing this insertion site is useful for expression of foreign antigen genes, as a vaccine against disease or as an expression vector to raise antibodies to the expressed foreign gene.

Example 30:

Recombinant swinepox virus expressing E coli β- galactosidase (lacZ) under the control of a synthetic early or synthetic late pox promoter.

Three recombinant swinepox viruses, S-SPV-056, S-SPV-

057, and S-SPV-058 expressing E coli /S-galactosidase (lacZ) under the control of a synthetic pox promoter,

LPl, LP2 , and EP1, respectively, have been constructed.

S-SPV-056 was derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing the homology vector 791-63.19 (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 _/S-galactosidase (BLUOGAL AND CPRG ASSAYS) . S-SPV-057 was derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing the homology vector 791-63.41 (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) . S-SPV-058 was derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing the homology vector 796-18.9 (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 were the recombinant viruses designated S-SPV-056, S-SPV-057 and S-SPV-058. The viruses were assayed for /S-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 foreign gene.

Recombinant swinepox virus expresses a foreign gene such as E. coli _/S-galactosidase in a human cell line but does not replicate in the human cell line. To optimize expression of the foreign gene, S-SPV-056, S- SPV-057 and S-SPV-058 are used to compare optimal expression levels of E. coli _/8-galactosidase under the control of early or late synthetic pox viral promoters . The human cell lines in which expression of recombinant swinepox virus has been detected include, but are not limited to 143B (osteosarcoma) , A431 (epidermoid carcinoma) , A549 (lung carcinoma), Capan-l (liver carcinoma) , CF500 (foreskin fibroblasts) , Chang Liver (liver) , Detroit (down's foreskin fibroblasts) , HEL-199 (embryonic lung) , HeLa (cervical carcinoma) , HEp-2 (epidermal larynx carcinoma) , HISM (intestinal smooth muscle) , HNK (neonatal kidney) , MRC-5 (embryonic lung) , NCI-H292 (pulmonary mucoepidermoid carcinoma) , OVCAR-3 (ovarian carcinoma) , RD (rhabdosarcoma) , THP (monocyte leukemia) , WIL2-NS (B lymphocyte line, non-secreting) , WISH (amnion) .

Example 31:

S-SPV-051

S-SPV-051 is a swinepox virus that expresses at least two foreign genes. The gene for E. coli _/S-galactosidase

(lacZ) and the gene for the bovine viral diarrhea virus glycoprotein 53 (g53) were inserted into the SPV 617-

48.1 ORF (a unique NotI restriction site has replaced a unique AccI restriction site) . The lacZ gene is under the control of the synthetic late promoter (LPl) , and the BVDV g53 gene is under the control of the synthetic late/early promoter (LP2EP2) .

S-SPV-051 was derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing the homology vector 783-39.2 (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 0-galactosidase (BLUOGAL AND CPRG ASSAYS) . The final result of red plaque purification was the recombinant virus designated S-SPV 051. This virus was assayed for _/S-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 foreign gene.

S-SPV-051 was assayed for expression of BVDV specific antigens using the BLACK PLAQUE SCREEN FOR FOREIGN GENE EXPRESSION IN RECOMBINANT SPV. A mouse monoclonal antibody to BVDV g53 was shown to react specifically with S-SPV-051 plaques and not with S-SPV-001 negative control plaques. All S-SPV-051 observed plaques reacted with the monoclonal antibody to BVDV g53 indicating that the virus was stably expressing the BVDV foreign gene. The assays described here were carried out in ESK-4 cells, indicating that ESK-4 cells would be a suitable substrate for the production of SPV recombinant vaccines.

To confirm the expression of the BVDV g53 gene product, cells were infected with S-SPV-051 and samples of infected cell lysates and culture supern atants were subjected to SDS polyacrylamide gel electrophoresis. The gel was blotted and analyzed using the WESTERN BLOTTING PROCEDURE. A mouse monoclonal antibody to BVDV g53 was used to detect expression of BVDV specific proteins. The cell lysate and supernatant from S-SPV-051 infected cells exhibited bands at 53 kd and higher indicating glycosylated and unglycosylated forms of the BVDV g53 protein.

S-SPV-051 is useful as a vaccine in cattle against BVDV infection and is useful for expression of BVDV glycoprotein 53.

Example 32:

S-SPV-044:

S-SPV-044 is a swinepox virus that expresses at least two foreign genes. The gene for E. coli /S-galactosidase (lacZ) and the gene for the infectious bursal disease virus (IBDV) polymerase protein were inserted into the 617-48.1 ORF (a unique NotI site has replaced a unique AccI restriction site) . The lacZ gene is under the control of the synthetic late promoter (LPl) , and the IBDV polymerase gene is under the control of the synthetic late/early promoter (LP2EP2) .

S-SPV-044 was derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing the homology vector 749-75.78 (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 /3-galactosidase (BLUOGAL AND CPRG ASSAYS) . The final result of red plaque purification was the recombinant virus designated S-SPV-044. This virus was assayed for _/S-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 foreign gene.

S-SPV-044 is useful for expression of IBDV polymerase protein. S-SPV-044 is useful in an in vi tro approach to a recombinant IBDV attenuated vaccine. RNA strands from the attenuated IBDV strain are synthesized in a bacterial expression system using T3 or T7 promoters

(pBlueScript plasmid; Stratagene, Inc.) to synthesize double stranded short and long segments of the IBDV genome. The IBDV double stranded RNA segments and S-

SPV-044 are transfected into CEF cells. The swinepox virus expresses the IBDV polymerase but does not replicate in CEF cells. The IBDV polymerase produced from S-SPV-044 synthesizes infectious attenuated IBDV virus from the double stranded RNA genomic templates . The resulting attenuated IBDV virus is useful as a vaccine against infectious bursal disease in chickens.

Example 33: S - SPV- 046 :

S-SPV-046 is a swinepox virus that expresses at least two foreign genes. The gene for E. coli /S-galactosidase (lacZ) and the gene for the feline immunodeficiency virus (FIV) gag protease (gag) were inserted into the

738-94.4 ORF (a 773 base pair deletion of the SPV OIL

ORF; Deletion of nucleotides 1669 to 2452, SEQ ID NO:

189) . The lacZ gene is under the control of the swinepox OIL promoter, and the FIV gag gene is under the control of the synthetic late/early promoter

(LP2EP2) .

S-SPV-046 was derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing the homology vector 761-75.B18 (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 _/S-galactosidase (BLUOGAL AND CPRG ASSAYS) . The final result of red plaque purification was the recombinant virus designated S-SPV 046. This virus was assayed for ■S-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 foreign gene.

To confirm the expression of the FIV gag gene product, cells were infected with S-SPV-046 and samples of infected cell lysates and culture supernatants were subjected to SDS polyacrylamide gel electrophoresis. The gel was blotted and analyzed using the WESTERN BLOTTING PROCEDURE. Feline anti-FIV (PPR strain) sera was used to detect expression of FIV specific proteins. The cell lysate and supernatant from S-SPV-046 infected cells exhibited bands at 26 kd and 17 kd which are the expected sizes of the processed form of the FIV gag protein. The recombinant swinepox virus expressed FIV gag protein is processed properly and secreted into the culture media.

S-SPV-048

S-SPV-048 is a swinepox virus that expresses at least two foreign genes. The gene for E. coli _/β-galactosidase (lacZ) and the gene for feline immunodeficiency virus (FIV) envelope (env) were inserted into the SPV 617 48.1 ORF (a unique NotI restriction site has replaced a unique AccI restriction site) . The lacZ gene is under the control of the synthetic late promoter (LPl) , and the FIV env gene is under the control of the synthetic late/early promoter (LP2EP2) .

S-SPV-048 was derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing the homology vector 781-84.Cll (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 _/S-galactosidase (BLUOGAL AND CPRG ASSAYS) . The final result of red plaque purification was the recombinant virus designated S-SPV 048. This virus was assayed for /S-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 foreign gene.

S-SPV-046 and S-SPV-048 are useful alone or in combination as a vaccine in cats against FIV infection and are useful for expression of the FIV env and gag proteins . A recombinant swinepox virus expressing both the FIV env and gag proteins is useful as a vaccine in cats against FIV infection.

Recombinant swinepox virus expressing human respiratory synctial virus F and G proteins is useful as a vaccine against the human disease.

Example 34

In Vitro Properties of Chicken IFN Expressed in Recombinant Pox viruses .

Growth properties of recombinant viruses in cell culture. Gowth properties of recombinant S-SPV-042 were not effected in embryonic swine kidney cells (ESK- 4) compared to wild-type swinpox virus.

Western blot analysis was performed on supernatants from cells infected with SPV/cIFN recombinant virus.

Rabbit and mouse antisera were raised against cIFN from concentrated SPV/cIFN infected supernatants and pre- cleared against ESK-4 cells infected with wild-type SPV in preparation for western analysis. Rabbit and mouse anti-cIFN antisera were reacted with denatured proteins on nitrocellulose from recombinant SPV/cIFN and SPV wild type virus infected supernatants. A reactive band with an estimated molecular weight size range of 17-20 kilodaltons was present in the SPV/cIFN lanes, and absent in the SPV wild type control lanes.

Effect of cIFN expressed in supernatants from SPV/cIFN (S-SPV-042) . FPV/cIFN. and FPV/cIFN/NDV infected cells on the growth of Vesicular Stomatis Virus. Virion cleared supernatants from SPV/cIFN, FPV/cIFN and FPV/cIFN/NDV infected cells were tested for the presence of viral inhibitory activity, results shown in Table 1. Briefly, CEF cells were incubated with serially diluted viral supernatants. Subsequently, 40,000 plaque forming units (pfu) /well of vesicular stomatitis virus (VSV) were added and 48 hours later, wells were scored for the presence of VSV cytopathic effect (CPE) . Recombinant viral supernatants containing cIFN were shown to inhibit VSV induced CPE, whereas, control viral supernatants did not. VSV induced cytopathic effect could be reversed in the presence of rabbit anti-cIFN sera.

Table 1.

Recombinant Viral cIFN Activity (units/ml! Supernatants .

SPV/IFN 2,500 000 SPV <100 FPV/IFN 250,000

FPV/cIFN/NDV 250, 000 FPV <100

One unit of cIFN activity is defined as the dilution of pox virus supernatant at which 100% VSV CPE was inhibited.

Effect of cIFN expressed from, supernatants of SPV/cIFN infected cells on herpes virus of turkeys .

Supernatant containing recombinant cIFN from ESK-4 cells infected with SPV/cIFN virus, was tested for its ability to inhibit the growth of herpes virus of turkeys (HVT) in CEF cells, results shown in Table 2. Briefly, serially diluted supernatants were incubated with CEF cells, and then subsequently infected with 100 pfu/well of wild-type HVT. Plaques were counted in all wells after 48 hours. It was shown that 10-100 units of cIFN activity inhibited plaque formation of HVT (100 pfu/well) . Supernatants from wild type SPV did not inhibit HVT plaque formation.

Table 2.

SPV/cIFN Supernatant Number of HVT plaques (units/ml^a)

0 99

1000 0 100 0 10 45

^a* One unit of cIFN activity is defined as the dilution of pox virus supernatant at which 100% VSV CPE was inhibited.

Induction of NO bv chicken macrophages after treatment with cIFN expressed in supernatants from SPV/cIFN infected cells.

HD 11 cells or bone marrow adherent cells were incubated with lOOOunit/ml of cIFN from SPV/cIFN supernatants, lipopolysaccharide (LPS) (6ng/ml) or with both cIFN and LPS, results shown in Table 3. After 24 hours, supernatant fluids were collected and nitrite levels were measured. These data demonstrate that cIFN expressed from SPV/cIFN supernatants has the ability to activate chicken macrophages in the presence of LPS. Table 3 .

Nitrite (micro/mol) levels following stimulation with :

Cell source LPS SPV/cIFN LPS + SPV/cIFN

HD11 10.76 6.4 35.29 BMAC 13.1 5.8 35.10

Conclusions:

1. Recombinant swinepox viruses express biologically active chicken interferon into the supernatants of infected cells, as measured by protection of CEF cells from VSV infection.

2. Chicken interferon expressed in supernatants from recombinant SPV/cIFN infected cells has been shown to protect CEF cells against infection with HVT in a dose dependent manner.

3. Chicken interferon expressed from SPV/cIFN acted synergistically with LPS to activate chicken macrophages as detected by nitric oxide induction.

4. The foregoing data indicate that recombinant swinepox viruses expressing chicken IFN may have beneficial applications as immune modulating agents in vi tro, in vivo and in ovo .

Example 35 As an alternative to the construction of a IBD vaccine using a viral vectored delivery system and/or subunit approaches, IBD virus RNA is directly manipulated re¬ constructing the virus using full length RNA derived from cDNA clones representing both the large (segment A) and small (segment B) double-stranded RNA subunits. Generation of IBD virus is this manner offers several advantages over the first two approaches. First, if IBD virus is re-generated using RNA templates, one is able to manipulate the cloned cDNA copies of the viral genome prior to transcription (generation of RNA) . Using this approach, it is possible to either attenuate a virulent IBD strain or replace the VP2 variable region of the attenuated vaccine backbone with that of virulent strains. In doing so, the present invention provides protection against the virulent IBDV strain while providing the safety and efficacy of the vaccine strain. Furthermore, using this approach, the present invention constructs and tests temperature sensitive IBD viruses generated using the RNA polymerase derived from the related birnavirus infectious pancreatic necrosis virus (IPNV) and the polyprotein derived from IBDV. The IPNV polymerase has optimum activity at a temperature lower than that of IBDV. If the IPNV polymerase recognizes the regulatory signals present on IBDV, the hybrid virus is expected to be attenuated at the elevated temperature present in chickens . Alternatively, it is possible to construct and test IBD viruses generated using the RNA polymerase derived from IBDV serotype 2 viruse and the polyprotein derived from IBDVserotype 1 virus.

cDNA clones representing the complete genome of IBDV (double stranded RNA segments A and B) is constructed, initially using the BursaVac vaccine strain (Sterwin Labs) . Once cDNA clones representing full length copies of segment A and B are constructed, template RNA is prepared. Since IBDV exists as a bisegmented double- stranded RNA virus, both the sense and anti-sense RΝA strands of each segment are produced using the pBlueScript plasmid; Stratagene, Inc.) . These vectors utilize the highly specific phage promoters SP6 or T7 to produce substrate amounts of RΝA in vi tro . A unique restriction endonuclease site is engineered into the 3' PCR primer to linearize the DΝA for the generation of run-off transcripts during transcription.

The purified RΝA transcripts (4 strands) are transfected into chick embryo fibroblasts (CEF) cells to determine whether the RΝA is infectious. If IBD virus is generated, as determined by black plaque assays using IBDV specific Mabs, no further manipulations are required and engineering of the vaccine strain can commence. The advantage of this method is that engineered IBD viruses generated in this manner will be pure and require little/no purification, greatly decreasing the time required to generate new vaccines. If negative results are obtained using the purified RΝA's, functional viral RΝA polymerase is required by use of a helper virus. Birnaviruses replicate their nucleic acid by a strand displacement (semi-conservative) mechanism, with the RΝA polymerase binding to the ends of the double-stranded RΝA molecules forming circularized ring structures (Muller & Νitschke, Virology 159, 174-177, 1987) . RΝA polymerase open reading frame of about 878 amino acids in swinepox virus is expressed and this recombinant virus (S-SPV-044) is used to provide functional IBDV RΝA polymerase in trans . Swinpox virus expressed immunologically recognizable foreign antigens in avian cells (CEF cells) , where there are no signs of productive replication of the viral vector. In the present invention the IBDV polymerase protein is expressed in the same cells as the transfected RΝA using the swinepox vector without contaminating the cells with SPV replication.

With the demonstration that IBD virus is generated in vitro using genomic RNA, an improved live attenuated virus vaccines against infectious bursal disease is developed. Using recombinant DNA technology along with the newly defined system of generating IBD virus, specific deletions within the viral genome, facilitating the construction of attenuated viruses are made. Using this technology, the region of IBDV responsible for virulence and generate attenuated, immunogenic IBDV vaccines are identified. The present invention provides a virulent IBD strain or replacement of the VP2 variable region of the attenuated vaccine backbone with that of a virulent strain, thus protecting against the virulent strain while providing the safety and efficacy of the vaccine strain.

Example 36

Effects of Rabbit anti-chicken interferon (cIFN) antibody on the growth of Herpes Virus of Turkeys.

Supernatants from SPV/cIFN (SPV 042) infected ESK-4 cells were harvested 48 hours after infection and then concentrated 5-10 times, by Centricon 10 columns (Amicon) . One ml of concentrated supernatant was injected into a rabbit 3 times, at 3 week intervals, and then bled. This rabbit antisera was then used in culture to study the effect of interferon on the growth of HVT. It was shown that anti-cIFN reverses the block to HVT (1:200) and VSV(1:80) growth induced by the addition of cIFN in plaque assays. Furthermore, it was shown that the addition of anti-cIFN (1:100) in the media of CEFs transiently transfected with sub- plaqueing levels of HVT viral DNA, enhances the formation of HVT plaques (200 plaques/well) . CEFs transfected with HVT DNA in the absence of anti-cIFN did not yield plaques.

HVT is highly susceptible to interferon produced from CEFs and that when cIFN is blocked, HVT growth is enhanced.

Applications include: (1) Use antibody to cIFN as an additive to increase HVT titers in vaccine stocks; (2) Use antibody to cIFN as an additive to facilitate the formation of new recombinant HVT viruses via cosmid reconstructions.

Example 37

S-SPV-063

S-SPV-063 is a swinepox virus that expresses at least two foreign genes. The gene for E. coli _/β-galactosidase

(lacZ) and the gene for swine influenza virus (SIV) NP

(HIND were inserted into the SPV 617 48.1 ORF (a unique NotI restriction site has replaced a unique AccI restriction site) . The lacZ gene is under the control of the synthetic late promoter (LPl) , and the SIV NP gene is under the control of the synthetic late/early promoter (LP2EP2) .

S-SPV-063 was derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing the homology vector 807-41.3 (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 _/S-galactosidase (BLUOGAL AND CPRG ASSAYS) . The final result of red plaque purification was the recombinant virus designated S-SPV 063. This virus was assayed for /S-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 foreign gene.

S-SPV-063 was assayed for expression of SIV specific antigens using the BLACK PLAQUE SCREEN FOR FOREIGN GENE

EXPRESSION IN RECOMBINANT SPV. Polyclonal swine anti-

SIV serum or a polyclonal goat anti-NP serum was shown to react specifically with S-SPV-063 plaques and not with S-SPV-001 negative control plaques. All S-SPV-063 observed plaques reacted with the swine anti-SIV serum or goat anti-NP serum indicating that the virus was stably expressing the SIV foreign gene. The assays described here were carried out in ESK-4 cells, indicating that ESK-4 cells would be a suitable substrate for the production of SPV recombinant vaccines.

To confirm the expression of the SIV NP gene products, cells were infected with S-SPV-063 and samples of infected cell lysates and culture supernatants were subjected to SDS polyacrylamide gel electrophoresis, The gel was blotted and analyzed using the WESTERN BLOTTING PROCEDURE. A polyclonal swine anti-SIV serum or a polyclonal goat anti-NP serum was used to detect expression of SIV specific proteins. The cell lysate and supernatant from S-SPV-063 infected cells exhibited bands corresponding to 56 kd, which is the expected size of the SIV NP protein.

S-SPV-063 is useful as a vaccine in swine against SIV infection and is useful for expression of SIV NP. S- SPV-063 is useful as a vaccine in combination with S- SPV-066 which expresses NA and S-SPV-065 which expresses SIV HA.

S-SPV-064

S-SPV-064 is a swinepox virus that expresses one foreign gene. The gene for E. coli _/β-glucuronidase (uidA) was inserted into the unique Xhol restriction site within the 6.9 kb SPV Hindlll J genomic fragment. The uidA gene is under the control of the synthetic late/early promoter (LP2EP2) . The Hindlll J genomic fragment contains part of the A50R ORF (aa 227 to 552) . The unique Xhol site is not within the A50R ORF. The Xhol site is 25 kb from the 3' end of the swinepox virus genome (62) .

S-SPV-064 was derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing the homology vector 807-42.28 and virus S-SPV-001 in the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT SPV.

Homology vector 807-42.28 was generated by insertion of a NotI fragment containing the LP2EP2 promoter uidA gene cassette from plasmid 551-47.23 (see Materials and Methods) into a NotI site (unique Xhol site converted to NotI by a DNA linker) in the SPV 6.9 kb Hindlll J fragment. The transfection stock was screened by the SCREEN FOR RECOMBINANT HERPESVIRUS EXPRESSING ENZYMATIC MARKER GENES. The final result of blue plaque purification is the recombinant virus designated S-SPV- 064. This virus is assayed for _/S-glucuronidase 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, plaques observed are blue indicating that the virus is pure, stable, and expressing the foreign gene. S-SPV-064 is purified and expresses the foreign gene, E. coli uidA, indicating that the Xhol site within the 6.9 kb Hindlll J fragment is a site non-essential for virus growth and a stable insertion site for foreign genes. Recombinant swinepox virus utilizing this insertion site is useful for expression of foreign antigen genes, as a vaccine against disease or as an expression vector to raise antibodies to the expressed foreign gene.

S-SPV-065

S-SPV-065 is a swinepox virus that expresses at least two foreign genes. The gene for E. coli _/β-galactosidase (lacZ) and the gene for swine influenza virus (SIV) HA (H1N1) were inserted into the SPV 617 48.1 ORF (a unique NotI restriction site has replaced a unique AccI restriction site) . The lacZ gene is under the control of the synthetic late promoter (LPl) , and the SIV HA gene is under the control of the synthetic late/early promoter (LP2EP2) .

S-SPV-065 was derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing the homology vector 807-84.8 (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 _/S-galactosidase (BLUOGAL AND CPRG ASSAYS) . The final result of red plaque purification was the recombinant virus designated S-SPV 065. This virus was assayed for /S-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 foreign gene. S-SPV-065 was assayed for expression of SIV specific antigens using the BLACK PLAQUE SCREEN FOR FOREIGN GENE EXPRESSION IN RECOMBINANT SPV. Polyclonal swine anti- SIV serum or a Polyclonal goat anti-HA serum was shown to react specifically with S-SPV-065 plaques and not with S-SPV-001 negative control plaques. All S-SPV-065 observed plaques reacted with the swine anti-SIV serum or the SIV foreign gene. The assays described here were carried out in ESK-4 cells, indicating that ESK-4 cells would be a suitable substrate for the production of SPV recombinant vaccines .

To confirm the expression of the SIV NP gene products, cells were infected with S-SPV-065 and samples of infected cell lysates and culture supernatants were subjected to SDS polyacrylamide gel electrophoresis. The gel was blotted and analyzed using the WESTERN BLOTTING PROCEDURE. A Polyclonal swine anti-SIV serum or a Polyclonal goat anti-HA serum was used to detect expression of SIV specific proteins. The cell lysate and supernatant from S-SPV-065 infected cells exhibited bands corresponding to 64 kd, which is the expected size of the SIV HA protein.

S-SPV-065 is useful as a vaccine in swine against SIV infection and is useful for expression of SIV HA. S- SPV-065 is useful as a vaccine in combination with S- SPV-066 which expresses NA and S-SPV-063 which expresses SIV NP.

S-SPV-066

S-SPV-066 is a swinepox virus that expresses at least two foreign genes. The gene for E. coli /S-galactosidase (lacZ) and the gene for swine influenza virus (SIV) NA (H1N1) were inserted into the SPV 617 48.1 ORF (a unique NotI restriction site has replaced a unique AccI restriction site) . The lacZ gene is under the control of the synthetic late promoter (LPl) , and the SIV NA gene is under the control of the synthetic late/early promoter (LP2EP2) .

S-SPV-066 was derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing the homology vector 807-84.35 (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 066. 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 foreign gene.

To confirm the expression of the SIV NA gene products, cells were infected with S-SPV-066 and samples of infected cell lysates and culture supernatants were subjected to SDS polyacrylamide gel electrophoresis. The gel was blotted and analyzed using the WESTERN BLOTTING PROCEDURE. A Polyclonal swine anti-SIV serum or a Polyclonal goat anti-NA serum was used to detect expression of SIV specific proteins. The cell lysate and supernatant from S-SPV-066 infected cells exhibited bands corresponding to 64 kd, which is the expected size of the SIV HA protein.

S-SPV-066 is useful as a vaccine in swine against SIV infection and is useful for expression of SIV NA. S-

SPV-066 is useful as a vaccine in combination with S- SPV-065 which expresses HA and S-SPV-063 which expresses SIV NP.

S-SPV-071

S-SPV-071 is a swinepox virus that expresses at least four foreign genes. The gene for E. coli β- galactosidase (lacZ) and the genes for swine influenza virus (SIV) HA (HlNl) and NA (HlNl) were inserted into the SPV 617 48.1 ORF (a unique NotI restriction site has replaced a unique AccI restriction site) . The lacZ gene is under the control of the synthetic late promoter (LPl) , and the SIV HA, and NA genes are under the control of the synthetic late/early promoter (LP2EP2) .

S-SPV-071 was derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing the homology vector817- 86.35 (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 _/S-galactosidase (BLUOGAL AND CPRG ASSAYS) . The final result of red plaque purification was the recombinant virus designated S-SPV 071. This virus was assayed for /S-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 foreign gene.

S-SPV-071 was assayed for expression of SIV specific antigens using the BLACK PLAQUE SCREEN FOR FOREIGN GENE EXPRESSION IN RECOMBINANT SPV. Polyclonal goat anti-HA serum was shown to react specifically with S-SPV-071 plaques and not with S-SPV-001 negative control plaques. All S-SPV-071 observed plaques reacted with the goat anti-HA serum indicating that the virus was stably expressing the SIV foreign gene. The assays described here were carried out in ESK-4 cells, indicating that ESK-4 cells would be a suitable substrate for the production of SPV recombinant vaccines.

To confirm the expression of the SIV HA and NA gene products, cells were infected with S-SPV-071 and samples of infected cell lysates and culture supernatants were subjected to SDS polyacrylamide gel electrophoresis. The gel was blotted and analyzed using the WESTERN BLOTTING PROCEDURE. A Polyclonal swine anti-SIV serum or a Polyclonal goat anti-HA serum was used to detect expression of SIV specific proteins. The cell lysate and supernatant from S-SPV-071 infected cells exhibited bands corresponding to 64 kd and 52 kd, which is the expected size of the SIV HA and NA protein.

S-SPV-071 is useful as a vaccine in swine against SIV infection and is useful for expression of SIV HA and NA. S-SPV-071 is useful as a vaccine in combination with S-SPV-063 which expresses SIV NP.

S-SPV-074

S-SPV-074 is a swinepox virus that expresses at least four foreign genes. The gene for E. coli β- glucuronidase (uidA) and the genes for swine influenza virus (SIV) HA (HlNl) and NA (HlNl) were inserted into the SPV 617 48.1 ORF (a unique NotI restriction site has replaced a unique AccI restriction site) . The uidA gene is under the control of the synthetic late/early promoter (LP2EP2) , and the SIV HA and NA genes are under the control of the synthetic late/early promoter (LP2EP2) .

S-SPV-074 was derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing the homology vector 817-14.2 (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 /S-galactosidase (BLUOGAL AND CPRG ASSAYS) . The final result of red plaque purification was the recombinant virus designated S-SPV 074. This virus was assayed for _/S-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 foreign gene.

S-SPV-074 was assayed for expression of SIV specific antigens using the BLACK PLAQUE SCREEN FOR FOREIGN GENE EXPRESSION IN RECOMBINANT SPV. Polyclonal swine anti- SIV serum was shown to react specifically with S-SPV- 074 plaques and not with S-SPV-001 negative control plaques. All S-SPV-074 observed plaques reacted with the goat anti-HA serum indicating that the virus was stably expressing the SIV foreign gene. The assays described here were carried out in ESK-4 cells, indicating that ESK-4 cells would be a suitable substrate for the production of SPV recombinant vaccines .

S-SPV-074 is useful as a vaccine in a swine against SIV infection and is useful for expression of SIV HA and Na. S-SPV-074 is useful as a vaccine in combination with S-SPV-063 which expresses SIV NP. S-SPV-063, -065, -066, -071, and -074, are useful alone or in combination as a vaccine in swine against swine influenza infection and are useful for expression of the SIV NP, HA, and NA proteins.

S-SPV-069

S-SPV-069 is a swinepox virus that expresses at least two foreign genes. The gene for E. coli _/β-galactosidase (lacZ) and the gene for human respiratory syncytial virus (HRSV) fusion (F) protein were inserted into the SPV 738-94.4 ORF (a 773 base pair deletion of the SPV OIL ORF; Deletion of nucleotides 1669 to 2452, SEQ ID NO: 189) . The lacZ gene is under the control of the swinepox P_0IL promoter and the HRSV F gene is under the control of the synthetic late/early promoter (LP2EP2) . S-SPV-069 was derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing the homology vector 810-29.A2 (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 /S-galactosidase (BLUOGAL AND CPRG ASSAYS) . The final result of red plaque purification was the recombinant virus designated S-SPV 069. This virus was assayed for _/S-galactosidase expression, purity, and insert stability by multiple passages monitored by the blue plague 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 foreign gene. S-SPV-069 was assayed for expression of HRSV specific antigens using the BLACK PLAQUE SCREEN FOR FOREIGN GENE EXPRESSION IN RECOMBINANT SPV. Monoclonal antibody 621 (Biodesign, Inc.) against HRSV F was shown to react specifically with S-SPV-069 plaques and not with S-SPV- 001 negative control plaques. All S-SPV-069 observed plaques reacted with the monoclonal antibody 621 indicating that the virus was stably expressing the PRV foreign gene. The assays described here were carried out in ESK-4 cells, indicating that ESK-4 cells would be a suitable substrate for the production of SPV recombinant vaccines .

S-SPV-078

S-SPV-078 is a swinepox virus that expresses at least two foreign genes. The gene for E. coli _/S-galactosidase

(lacZ) and the gene for human respiratory syncytial virus (HRSV) attachment (G) protein were inserted into the SPV 617 48.1 ORF (a unique NotI restriction site has replaced a unique AccI restriction site) . The lacZ gene is under the control of the synthetic late/early promoter (LP2EP2) , and the HRSV G gene is under the control of the synthetic late/early promoter (LP2EP2) . S-SPV-078 was derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing the homology vector 822-52G.7 (see Materials and Methods) and virus S-SPV- 001 in the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT SPV. The transfection stock is screened by the SCREEN FOR RECOMBINANT SPV EXPRESSING _/β-galactosidase (BLUOGAL AND CPRG ASSAYS) . The final result of red plaque purification is the recombinant virus designated S-SPV-078. This virus is assayed for _/S-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 are blue indicating that the virus is pure, stable, and expressing the foreign gene.

S-SPV-069 and S-SPV-078 are useful individually or in combination as a vaccine in swine against human respiratory syncytial virus infection and are useful for expression of HRSV F and G genes.

HOMOLOGY VECTOR 810-29.A2. The plasmid 810-29.A2 was constructed for the purpose of inserting foreign DNA into SPV. It incorporates an E. coli _/β-galactosidase (lac Z) marker gene and a human respiratory syncytial virus (HRSV) fusion (F) gene flanked by SPV DNA. Upstream of the foreign gene is an approximately 855 base pair fragment of SPV DNA. Downstream of the foreign genes is an approximately 1113 base pair fragment of SPV DNA. When the 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 swinepox virus OIL gene promoter and the HRSV F gene is under the control of the late/early promoter (LP2EP2) . The homology vector was constructed utilizing standard recombinant DNA techniques (22, 30) , by joining restriction fragments from the following sources with the synthetic DNA sequences. The plasmid vector was derived from an approximately 2519 base pair Hindlll to SphI restriction fragment of pSP65 (Promega) . Fragment 1 is an approximately 855 base pair sub-fragment of the SPV Hindlll restriction fragment M (23) synthesized by polymerase chain r e a c t i on u s i ng DNA p r i me r s 5 ' GAAGCATGCCCGTTCTTATCAATAGTTTAGTCGAAAATA-3 ' (SEQ ID NO: 185) and 5' -CATAAGATCTGGCATTGTGTTATTATACTAACAAAAATAAG- 3' (SEQ ID NO: 186) to produce an 855 base pair fragment with SphI and Bglll ends. Fragment 2 is a 3002 base pair BamHI to PvuII fragment derived from plasmid pJF751 (49) containing the E. coli lacZ gene. Fragment 3 is an approximately 1728 base pair EcoRI restriction fragment synthesized by reverse transcriptase and polymerase chain reaction (PCR) (15, 42) using RNA from the HRSV Strain A2 (ATCC VR-1302) . The primer (5' GCCGAATTCGCTAATCCTCAAAGCAAATGCAAT- 3' ,-4/95.23) (SEQ ID NO:) synthesizes from the 5' end of the HRSV F gene, introduces an EcoRI site at the 5' end of the gene and an ATG start codon. The primer (5'- GGTGAATTCTTTATTTAGTTACTAAATGCAATATTATTT-3' ; 4/95.24) (SEQ ID NO:) synthesizes from the 3' end of the HRSV F gene and was used for reverse transcription and polymerase chain reaction. The PCR product was digested with EcoRI to yield a fragment 1728 base pairs in length corresponding to the HRSV F gene. Fragment 4 is an approximately 1113 base pair subfragment of the SPV Hindlll fragment M synthesized by polymerase chain re a c t i on u s i ng DNA pr ime r s 5 ' - CCGTAGTCGACAAAGATCGACTTATTAATATGTATGGGATT-3' (SEQ ID N O : 1 8 7 ) a n d 5 ' GCCTGAAGCTTCTAGTACAGTATTTACGACTTTTGAAAT-3' (SEQ ID NO: 188) to produce an 1113 base pair fragment with Sail and Hindlll ends.

HOMOLOGY VECTOR 822-52G.7. The plasmid 822-52G.7 was constructed for the purpose of inserting foreign DNA into SPV. It incorporates an E . coli _/S-galactosidase (lacZ) marker gene and the human respiratory syncytial virus (HRSV ) attachment (G) gene flanked by SPV DNA.

Upstream of the foreign genes is an approximately 1484 base pair fragment of SPV DNA. Downstream of the foreign genes is an approximately 2149 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 _/S-galactosidase (lacZ) marker gene is under the control of a synthetic late/early pox promoter (LP2EP2) and the HRSV G gene is under the control of a synthetic late/early pox promoter (LP2EP2) .It was constructed utilizing standard recombinant DNA techniques (22 and 30) , by joining restriction fragments from the following sources. The plasmid vector was derived from an approximately 2972 base pair Hindlll to BamHI restriction fragment of pSP65 (Promega) . Fragment 1 is an approximately 1484 base pair AccI to Bglll restriction sub-fragment of the SPV Hindlll fragment M (23) . Fragment 2 is an approximately 3006 base pair BamHI to PvuII restriction fragment of plasmid pJF751 (11) . Fragment 3 is an approximately 899 base pair EcoRI restriction fragment synthesized by reverse transcriptase and polymerase chain reaction (PCR) (15, 42) using RNA from the HRSV Strain A2 (ATCC VR-1302) . The primer (5' GCCGAATTCCAAAAACAAGGACCAACGCAC- 3';4/95.25) (SEQ ID NO:) synthesizes from the 5' end of the HRSV F gene, introduces an EcoRI site at the 5' end of the gene and an ATG start codon. The primer (5'- GCCGAATTCACTACTGGCGTGGTGTGTTG-3' ; 4/95.26) (SEQ ID NO: ) synthesizes from the 3' end of the HRSV G gene and was used for reverse transcription and polymerase chain reaction. The PCR product was digested with EcoRI to yield a fragment 899 base pairs in length corresponding to the HRSV G gene. Fragment 4 is an approximately 2149 base pair Hindlll to AccI restriction sub-fragment of the SPV Hindlll restriction fragment M (23) .

HOMOLOGY VECTOR 807-41.3. The plasmid 807-41.3 was used to insert froeign DNA into SPV. It incorporates an E. coli B- galactosidase (lacZ) marker gene and the swine influenza virus (SIV) nucleoprotein (NP) gene flanked by SPV DNA. When this plasmid was used according to theHOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING

RECOMBINANT SPV a virus containing DNA coding for the foreign genes results. NOte that the B galactosidase

(lacZ) marker gene is under the control of a synthetic late pox promoter (LPl) and the SIV NP gene is under the control of a synthetic late/early pox prometer (LP2EP2) . The homology vector was constructed utilizing standard recombinant DNA techniques (22 and 30) , by joining restriction fragments from the following sources with the appropriate synthetic DNA sequences. The plasmid vector was derived from an approximately 2972 base pair Hindiii to BAm HI restriction fragment of pSP64 (Promega) . Fragment 1 is an approximately 1484 base Bglii to AccI restriction sub-fragment of the SPV Hindlll fragment M (23) . Fragment 2 is an approximately 1501 base pair EcoRI to EcoRI fragment of the SIV NP gene synthesized by reverse transcription (RT) and polymerase chain reaction (PCR)

(15,42) using RNA from the SIV HlNl strain (NVSL) . The primer (5' CATGAATTCTCAAGGCACCAAACGATCATATGAAC-3 ' ;

6/95.13) (SEQ ID NO:) synthesizes from the 5' end of the SIV NP gene and introduces an EcoRI site at the 5' end of the gene. The primer (5' - ATTTGAATTCAATTGTCATACTCCTCTGCATTGTCT-3';6/95.14) (SEQ ID NO:) synthesizes from the 3' end of the SIV NP gene, introduces an EcoRI site st the 3' end of the gene, and was used for reverse transcription and polymarase chain reaction. The PCR product was digested with EcoRI to yield a fragment 1501 base pairs in length corresponding to the SIV NP gene. Fragment 3 is approximately 3010 base pair BamHI ro PuvII restriction fragmebt of plsmid pJF751 (11) . Fragent 4 is approximately 2149 base pair AccI to Hindlll restriction sub-fragment of the SPV Hind III restriction fragment M (23)

HOMOLOGY VECTOR 807-84.8. THe plasmid 807-84.8 was used to insert foreign DNA into SPV. It incorporates an E. coli B- galactosidase (lacZ) marker gene and the swine influenza virus (SIV) hemmagglutinin (HA) gene flanked by SPV DNA. When this plasmid was used according to the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GANERATING RECOMBINANT SPV a virus containing DNA coding for the foreign genes results. Note that the B galactosidase (lacZ) marker gene is under the control of a synthetic late pox promoter (LPl) and the SIV HA gene is under the control of a synthetic late/early promoter (LP2EP2) . The homology vector was constructed utilizing standard recombinant DNA techniques (22 and 30) , by joining restricting fragments from the following sources with the appropriate synthetic DNA sequences . The plasmid vector was derived from an approximately 2972 base pair Hindlll to BamHI restriction fragment of pSP64 (Promega) . Fragment 1 is an approximately 1484 base pair Bglll to AccI restriction sub-fragment of the SPV HindiII fragment M (23) . Fragment 2 is an approximately 1721 base pair BamHI to BamHI fragment of the SIV HA gene synthesized by reverse transcription

(RT) and polymerase chain reaction (PCR) (15,42) using

RNA from the SIV HlNl strain (NVSL) . The primer (5'

CCGAGGATCCGGCAATACTATTAGTCTTGCTATGTACAT-3 ' ; 6/95.5)

(SEQ ID NO:) synthesizes from the 5' end of the SIV HA gene and introduces an BamHI site at the 5; end of the g e n e . T h e p r i m e r ( 5 ' - CTCTGGATCCTAATTTAAATACATATTCTGCACTGTS-3'; 6/95.6) (SEQ ID NO:) synthesizes from the 3' end of the SIV HA gene, introduces a BAm HI site at the 3' end of the gene, and was used for the reverse transcription and polymerase chain reaction. The PCR product was digested with EcoRI to yield a fragment 1721 base pairs in length corresponding to the SIV HA gene. Fragment 3 is an approximately 3010 base pair BamHI ot PvuII restriction fragment of plasmid pJF751 (11) . Fragment 4 is an approximately 2149 base pair AccI to fragmeny M (23) .

HOMOLOGY VACTOR 807-84.35. The plasmid 807-84.35 was used to insert foreign DNA into SPV> It incorporates an E_> coli B-galactosidase (lacZO marker gene and the swine influenza virus (SIV) neuraminidase (NA) gene flanked by SPV DNA. When this PROCEDURE FRO GENERATING RECOMBINANT SPV a virus containing DNA coding for the foreign genes results. Note that the B galactosidase

(lacZ) marker gene is under the comtrol of a synthetic late pox promoter (LPl) and the SIV NA gene is under the comtrol of a synthetic late/early pox promoter

(LP2EP2) . The homology vector was constructed utilizing standard recombinant DNA techniques (22 and

30) . by joinung restricting fragments from the following sources with the appropriate synthetic DNA sequences . The plasmid vector was derived from an approximately 2972 base pair Hindlll to BamHI restriciton fragment of pSP64 (Promega) . Fragment 1 is an approximately 1484 base pair Bglll to AccI restriction sub-fragment of the SPV Hindlll fragment M (23) . Fragment 2 is an approximately 1414 base pair EcoRI ti Bglll fragment of the SIV NA gene synthesized by reverse transcription (RT) and polymerase chain reaction (PCR) (15,42) using RNA from the SIV HlNl s t r a i n ( NVS L ) . TH e p r i m e r ( 5 ' AATGAATTCAAATCAAAAAATAATAACCATTGGGTCAAT-3' ; 6/95.12)

(SEQ ID NO:) synthesizes from the 5' end of the SIV NA gene and introduces an EcoRI site at the 5' end of the g e n e . T h r e p r i m e r ( 5 ' - GGAAGATCTACTTGTCAATHHTHAATGGCAGATCAG-3'; 6/95.13) (SEQ ID NO:) synthesizes from the 3' end of the SIV NA gene, introduces an Bglll site at the 3' end of the gene, and was used for reverse transcription and polymerase chain recation. The PCR product was digested with EcoRI to yield a fragment 1414 base pairs in length corresponding to the SIV NA gene. Fragment 3 is an approximately 3010 base pair BamHI to PvuII restriciton fragment of plasmid pJF751 (11) . Fragment 4 is an approximately 2149 base pair AccI to Hindlll restriciton sub-fragment of the SPV Hindlll restriction fragment M (23) . HOMOLOGY VECTOR 807-86.35. The plasmid 807-86.35 was used to insert foreign DNA into SPV. It incorporates an E> coli B- galactosidase (lacZ) marker gene and the swine influenza virus (SIV) hemagglutinin (HA) and neuraminidase (NA) gene flanked by SPV DNA> When this plasmid was used according to the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT SPV a virus containing DNA coding for the foreign genes results. Note that the B galactosidase (lacZ) marker gene is under the control of a synthetic late pox promoter (LPl) and the SIV NA and HA genes are each under the control of a synthetic late/early pox promoter (LP2EP2) . The homology vector was constructed utilizing standard recombinant DNA techniques (22 and 30) , by joining restriction fragments from the following sources with the appropriate synthetic DNA sequences. The plasmid vector was derived from an approximately 2972 base pair Hindi11 to BamHI restriction fragment of pSP64 (Promega) . Fragment 1 is approximately 1484 base pair Bglll to AccI restriction sub-fragmentof the SPV Hindlll fragment M (23) . Fragment 2 is an approximately 1721 base pair BamHI to BamHI fragment of the SIV HA gene synthesized by reverse transcription (RT) and polymerase chain reaction (PCR) (15,42) using RNA from the SIV HlNl strain (NVSL) . The primer ( 5 ' - CCGAGGATCCGGCAATACTATTAGTCTTGCTATGTACAT-3' ;6/95.5) (SEQ ID NO:) synthesizes from the 5' end of the SIV HA gene and introduces an Bam HI site at the 5' end of the g e n e . T h e p r i m e r ( 5 ' -

CTCTGGGATCCTAATTTTAAATACATATTCTGCACTGTA-3 ' ; 6/95.6)

(SEQ ID NO:) synthesizes from the 3' end of the SIV HA gene, introduces an BamHI site at the 3' end of the gene, and was used for reverse transcription and polymerase chain reaction. The PCR product was digested with EcoRI to yield a fragment 1721 base pairs in length corresponding to the SIV HA gene. Fragment 3 is an approximately 1414 base pair EcoRI to Bglll fragment of the SIV NA gene synthesized by reverse transcription

(RT) and polymerase chain reaction (PCR) (15,42) using

RNA from the SIV HlNl strain (NVSL) . The primer (5' AATGAATTCAAATCAAAAAATAATAACCATTGGGTCAAT-3 ' ; 6/95.12)

(SEQ ID NO:) synthesizes from the 5' end of the SIV NA gene and introduces an EcoRI site at the 5' end of the g e n e . T h e p r i m e r ( 5 ' -

GGAAGATCTACTTGTCAATGGTGAATGGCAGATCAG-3'; 6/95.13) (SEQ ID NO:) synthesizes from the 3' end of the SIV NA gene, introduces an Bglll site at the 3' end of the gene, and was used for reverse transcription and plymerase chain reaction. The PCR product was digested with EcoRI to yield a fragment 1414 base pairs in length corresponding to the SIV NA gene. Fragment 4 is an approximately 3010 base pair BamHI to PvuII restriction fagment of plasmid pJF751 (11) . Fragment 5 is an approximately 2149 base pair AccI to Hindlll restriction sub-fragment of the SPV Hindlll restriction fragment M (23) .

HOMOLOGY VECTOR 817-14.2. The plasmid 817-14.2 was used to insert foreign DNA into SPV. It incorporates an E. coli B- glucuronidase (uidA) marker gene and the swine influenza virus (SIV) henagglutinin (HA) and neuraminidase (NA) gene flanked by SPV DNA, When theis poasmid was used according to the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT SPV a virus containing DNA coding for the foreign genes results. Note that the B glucuronidase (uidA) marker gene is under the control of a synthetic late/early pox pomoter (LP2EP2) and the SIV NA and HA genes are each under the control of a synthetic late/early pox promoter (LP2EP2) . The homology vector was constructed utilizing standard recombinant DNA techniques (22 and 30) , by joining restriction fragments from the following sources with the appropriate synthetic DNA sequences. The plasmid vector was derived from an approximately 2972 base pair Hindlll to BamHI restriction fragment of pSP64 (Promega) . Fragment 1 is an approximatelyl484 base pair Bglll to AccI restriction subfragment fo the SPV Hindlll fragment M (23) . Fragment 2 is an approximately 1721 base pair BamHI to BamHI fragment of the SIV HA gene synthesized by reverse transcription (RT) and polymerase chain reaction (PCR) (15,42) using RNA from the SIV HlNl st rain ( NVSL ) . The prime r ( 5 ' CCGAGGATCCGGCAATACTATTAGTCTTGCTATGTACAT-3' ;.6/95.5) (SEQ ID NO:) synthesizes from the 5' end of the SIV HA gene and introduces an BamHI site at the 5' end of the gene. Theprimer (5' -CTCTGGGATCCTAATTTTAAATACATATTCTGCACTGTA- 3'; 6/95.6) (SEQ ID NO:) synthesizes from the 3' end of the SIV HA gene, introduces an BamHI site at the 3' end of the gene, and was used for reverse transcription and polymerase chain reaction. The PCR product was digested with EcoRI to yield a fragment 1721 base pairs in length corresponding to the SIV HA gene. Fragment 3 is an approximately 1414 base pair EcoRI to Bglll fragment of the SIV NA gene synthesized by reverse transcription (RT) and polymerase chain reaction (PCR) (15,42) using RNA fro the SIV HlNl strain (NVSL) . The primer (5' AATGAATTCAAATCAAAAAATAATAACATTGGGTCAAT-3',-6/95.12) (SEQ ID NO:) synthesizes from the 5' end of the SIV NA gene and introduces an EcoRI site at the 5' end of the gene. Theprimer (5' -GGAAGATCTACTTGTCAATGGTGAATGGCAGATCAG-3' ; 6/95.13) (SEQ ID NO:) synthesizes from the 3' end of the SIV NA gene, introduces an Bglll site at the 3' end of the gene, and was used for reverse transcription and polymerase chainreaction. The PCR product was digested with EcoRI to yield a fragment 1414 base pairs in length corresponding to the SIV NA gene. Fragment 4 is an approximately 1823 vase pair NotI restriction fragment of plasmid pRAJ260 (Clonetech) . Fragment 5 is an approximately 2149 base pair AccI to Hindlll restriction sub-fragment of the SPV Hindlll restriction fragment M (23) .

PRRS HOMOLOGY VECTORS CONTAINING SINGLE OR MULTIPLE PRRS GENES (ORF2, ORF3, ORF4, ORF5, ORF6, or ORF7: The

PRRS homology vector is constructed for the purpose of inserting foreign DNA into SPV. It incorporates an E. coli /S-galactosidase (lac Z) marker gene and a porcine reproductive and respiratory syndrome virus (PRRS) ORF2, ORF3, ORF4, ORF5, ORF6, or ORF7 gene flanked by SPV DNA. Upstream of the foreign gene is an approximately 855 base pair fragment of SPV DNA. Downstream of the foreign genes is an approximately 1113 base pair fragment of SPV DNA. When the 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 swinepox virus OIL gene promoter and the PRRS gene is under the control of the late/early promoter (LP2EP2) . The homology vector was constructed utilizing standard recombinant DNA techniques (22, 30), by joining restriction fragments from the following sources with the synthetic DNA sequences. The plasmid vector was derived from an approximately 2519 base pair HINDIII to SphI restriction fragment of pSP65 (Promega) . Fragment 1 is an approximately 855 base pair sub-fragment of the SPV Hindlll restriction fragment M (23) synthesized by polymerase chain reaction using DNA primers 5'

GAAGCATGCCCGTTCTTATCAATAGTTTAGTCGAAAATA-3' (SEQ ID NO: 185) and 5' -CATAAGATCTGGCATTGTGTTATTATACTAACAAAAATAAG- 3' (SEQ ID NO: 186) to produce an 855 base pair fragment with SphI and Bglll ends. Fragment 2 is a 3002 base pair BamHI to PvuII fragment derived from plasmid pJF751 (49) containing the E. coli lacZ gene. Fragment 3 is an EcoRI to BamHI restriction fragment synthesized by reverse transcription and polymerase chain reaction (PCR) using genomic RNA from a U.S. Isolate of PRRS obtained from the NVSL (Reference strain) . Each homology vector contains one or multiple of the PRRS virus 0RF2 through 7. To synthesize PRRS 0 R F 2 , t h e p r i m e r ( 5 ' AATGAATTCGAAATGGGGTCCATGCAAAGCCTTTTTG-3' ; 1/96.15) (SEQ ID NO:) synthesizes from the 5' end of the PRRS 0RF2 gene, introduces an EcoRI site at the 5' end of the g e n e . T h e p r i m e r ( 5 ' - CAAGGATCCCACACCGTGTAATTCACTGTGAGTTCG-3'; 1/96.16) (SEQ ID NO.) is used for reverse transcription and PCR and synthesizes from the 3' end of the PRRS 0RF2 gene. The PCR product was digested with EcoRI and BamHI to yield a fragment 771 base pairs in length corresponding to the PRRS ORF2 gene. To synthesize PRRS ORF3, the primer (5' TTCGAATTCGGCTAATAGCTGTACATTCCTCCATATTT-3' ; 1/96.7) (SEQ ID NO:) synthesizes from the 5' end of the PRRS 0RF3 gene, introduces an EcoRI site at the 5' end of the gene . The primer ( 5 ' - GGGGATCCTATCGCCGTACGGCACTGAGGG-3' ; 1/96.8) (SEQ ID NO: ) is used for reverse transcription and PCR and synthesizes from the 3' end of the PRRS ORF3 gene. To synthesize PRRS 0RF4 , the primer (5' CCGAATTCGGCTGCGTCCCTTCTTTTCCTCATGG-3'; 1/96.11) (SEQ ID NO:) synthesizes from the 5' end of the PRRS ORF4 gene, introduces an EcoRI site at the 5'- CTGGATCCTTCAAATTGCCAACAGAATGGCAAAAAGAC-3'; 1/96.12) (SEQ ID NO.) is used for reverse transcription and PCR and synthesizes from the 3' end of the PRRS ORF4 gene. The PCR product was digested with EcoRI and BamHI to yield a fragment 537 base pairs in length corresponding to the PRRS ORF4 gene. To synthesize PRRS 0RF5, the primer (5' TTGAATTCGTTGGAGAAATGCTTGACCGCGGGC-3' ; 1/96.13) (SEQ ID NO:) synthesizes from the 5' end of the PRRS 0RF5 gene, introduces an EcoRI site at the 5' end of the gene. The primer (5'- GAAGGATCCTAAGGACGACCCCATTGTTCCGCTG-3' ; 1/96.14) (SEQ ID NO:) is used for reverse transcription and PCR and synthesizes from the 3' end of the PRRS 0RF5 gene. The PCR product was digested with EcoRI and BamHI to yield a fragment 603 base pairs in length corresponding to the PRRS ORF5 gene. To synthesize PRRS ORF6, the primer (5' CGGGAATTCGGGGTCGTCCTTAGATGACTTCTGCC-3 ' ; 1/96.17) (SEQ ID NO:) synthesizes from the 5' end of the PRRS ORF6 gene, introduces an EcoRI site at the 5' end of the gene. The primer (5'

GCGGATCCTTGTTATGTGGCATATTTGACAAGGTTTAC-3' ; 1/96.18)

(SEQ ID NO:) is used for reverse transcription and PCR and synthesizes from the 3' end of the PRRS ORF6 gene.

The PCR product was digested with EcoRI and BamHI to yield a fragment 525 base pairs in length corresponding to the PRRS ORF6 gene. To synthesize PRRS ORF7, the primer (5' GTCGAATTCGCCAAATAACAACGGCAAGCAGCAGAAG-3 ' ; 1/96.19) (SEQ ID NO:) synthesizes from the 5' end of the PRRS ORF7 gene, introduces an EcoRI site at the 5' end of the gene. The primer (5'- CAAGGATCCCAGCCCATCATGCTGAGGGTGATG-3' ; 1/96.20) (SEQ ID NO:) is used for reverse transcription and PCR and synthesizes from the 3' end of the PRRS ORF7 gene. Fragment 4 is an approximately 1113 base pair subfragment of the SPV Hindlll fragment M synthesized by polymerase chain reaction using DNA primers 5'- CCGTAGTCGACAAAGATCGACTTATTAATATGTATGGGATT-3' (SEQ ID NO: 187) and 5' GCCTGAAGCTTCTAGTACAGTATTTACGACTTTTGAAT- 3' ) SEQ ID NO: 188) to produce an 1113 base pair fragment with Sail and Hindlll ends.

Recombinant swinepox virus expressing pseudorabies genes

S-SPV-076 is a swinepox virus that expresses at least three foreign genes. The gene for E. coli β- galactosidase (lacZ) and the genes for pseudorabies virus (PRV) gD and gl were inserted into the SPV 617 48.1 ORF (a unique NotI restriction site has replaced a unique AccI restriction site) . The lacZ gene is under the control of the synthetic late promoter (LPl) , and the PRV gD and gl genes are under the control of the synthetic late/early promoter (LP2EP2) .

S-SPV-077 is a swinepox virus that expresses at least two foreign genes. The gene for E. coli β- galactosidase (lacZ) and the gene for pseudorabies virus (PRV) gl were inserted into the SPV 617 48.1 ORF

(a unique NotI restriction site has replaced a unique

AccI restriction site) . The lacZ gene is under the control of the synthetic late promoter (LPl) , and the PRV gl gene is under the control of the synthetic late/early promoter (LP2EP2) .

S-SPV-079 is a swinepox virus that expresses at least two foreign genes. The gene for E. coli β- galactosidase (lacZ) and the genes for pseudorabies virus (PRV) gB were inserted into the SPV 617 48.1 ORF (a unique NotI restriction site has replaced a unique AccI restriction site) . The lacZ gene is under the control of the synthetic late promoter (LPl) , and the PRV gB gene are under the control of the synthetic late/early promoter (LP2EP2) .

S-SPV-076, S-SPV-077, and S-SPV-079 were derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing a homology vector and virus S-SPV-001 in the HOMOLOGOUS RECOMBINATION PROCEDURE FOR GENERATING RECOMBINANT SPV. The transfection stock were 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-076, S-SPV-077, and S-SPV-079. The viruses were assayed for /S-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 foreign gene.

S-SPV-076, S-SPV-077, S-SPV-079 have been tested by BLACK PLAQUE ASSAY and WESTERN BLOT for expression of the PRV glycoproteins.

S-SPV-076, S-SPV-077, and S-SPV-079 are useful as a vaccine in swine against PRV infection and is useful for expression of PRV gD, gl or gB. S-SPV-071 is useful as a vaccine in combination with a recombinant swinepox virus which expresses PRV gC, such as S-SPV- 011, S-SPV-012, or S-SPV-013.

Example 38

Recombinant swinepox virus expressing PRRS genes 0RF2, ORF3, ORF4. 0RF5. and ORF6

S-SPV-080 is a swinepox virus that expresses at least two foreign genes. The gene for E. coli /S-galactosidase (lacZ) and the gene for porcine reproductive and respiratory syndrome virus (PRRS) ORF2 were inserted into the SPV 738-94.4 ORF (a 773 base pair deletion of the SPV OIL ORF; Deletion of nucleotides 1669 to 2452, SEQ ID NO: 189) . The lacZ gene is under the control of the swinepox P_0IL promoter and the PRRS ORF2 gene is under the control of the synthetic late/early promoter (LP2EP2) .

S-SPV-081 is a swinepox virus that expresses at least two foreign genes. The gene for E. coli /3-galactosidase (lacZ) and the gene for porcine reproductive and respiratory syndrome virus (PRRS) ORF3 were inserted into the SPV 738-94.4 ORF (a 773 base pair deletion of the SPV OIL ORF; Deletion of nucleotides 1669 to 2452, SEQ ID NO: 189) . The lacZ gene is under the control of the swinepox P_0IL promoter and the PRRS ORF3 gene is under the control of the synthetic late/early promoter (LP2EP2) .

S-SPV-082 is a swinepox virus that expresses at least two foreign genes. The gene for E. coli /S-galactosidase (lacZ) and the gene for porcine reproductive and respiratory syndrome virus (PRRS) ORF4 were inserted into the SPV 738-94.4 ORF (a 773 base pair deletion of the SPV OIL ORF; Deletion of nucleotides 1669 to 2452, SEQ ID NO: 189) . The lacZ gene is under the control of the swinepox P_0IL promoter and the PRRS ORF4 gene is under the control of the synthetic late/early promoter (LP2EP2) .

S-SPV-083 is a swinepox virus that expresses at least two foreign genes. The gene for E. coli _/S-galactosidase

(lacZ) and the gene for porcine reproductive and respiratory syndrome virus (PRRS) ORF5 were inserted into the SPV 738-94.4 ORF (a 773 base pair deletion of the SPV OIL ORF; Deletion of nucleotides 1669 to 2452, SEQ ID NO: 189) . The lacZ gene is under the control of the swinepox P_0IL promoter and the PRRS ORF5 gene is under the control of the synthetic late/early promoter

(LP2EP2) .

S-SPV-084 is a swinepox virus that expresses at least two foreign genes. The gene for E. coli _/β-galactosidase (lacZ) and the gene for porcine reproductive and respiratory syndrome virus (PRRS) ORF6 were inserted into the SPV 738-94.4 ORF (a 773 base pair deletion of the SPV OIL ORF; Deletion of nucleotides 1669 to 2452,

SEQ ID NO: 189) . The lacZ gene is under the control of the swinepox P_0IL promoter and the PRRS ORF6 gene is under the control of the synthetic late/early promoter

(LP2EP2) .

S-SPV-085 is a swinepox virus that expresses at least two foreign genes. The gene for E. coli _/β-galactosidase (lacZ) and the gene for porcine reproductive and respiratory syndrome virus (PRRS) 0RF7 were inserted into the SPV 738-94.4 ORF (a 773 base pair deletion of the SPV OIL ORF; Deletion of nucleotides 1669 to 2452, SEQ ID NO: 189) . The lacZ gene is under the control of the swinepox P_0IL promoter and the PRRS ORF6 gene is under the control of the synthetic late/early promoter (LP2EP2) . S-SPV-080, S-SPV-081, S-SPV-082, S-SPV-083, S-SPV-084, S-SPV-085 were derived from S-SPV-001 (Kasza Strain) . This was accomplished utilizing the homology vector Materials and Methods (PRRS HOMOLOGY VECTORS) 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-080, S-SPV-081, S-SPV-082, S- SPV-083, S-SPV-084, S-SPV-085. 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 foreign gene.

S-SPV-080, S-SPV-081, S-SPV-082, S-SPV-083, S-SPV-084, S-SPV-085 are useful individually or in combination as vaccines in swine against PRRS infection and are useful for expression of PRRS ORF2, ORF3, ORF4 , ORF5, ORF6 and ORF7.

Example 39

The following experiment was performed to determine the ability of swinepox virus to infect human cells in culture and express a foreign DNA such as lacZ.

S-SPV-003 was adsorbed to the human cell lines listed in the Table below at an MOI=0.1 for 2 to 3 hours. Cells were rinsed three times with PBS, growth media was added, and cells were incubated at 37 for four days. Cells were harvested and a lysate prepared in 200 microliters of PBS by freeze/thaw three times. Cell debris was pelleted, and 10 microliters of supernatant was assayed for -galactosidase activity by ONPG assay at 37 for 1 1/2 hours. The table shows the results of infection of various human cell lines with S-SPV-003 and the relative levels of cytopathetic effect and expression of lacZ.

The results show that various human cell lines vary in the ability to take up S-SPV-003 and express lacZ. CPE was minimal in all cases and did not result in viral replication. One exception A549 cells which did show some rounding of cells and lifting off the plate in one instance, and another instance of a ten-fold increase in titer during passage suggesting limited viral replication. Several cell lines how significant lacZ activity with no cytopathic effect.

Different pox promoters express lac Z from recombinant swinepox virus in a number of human cell lines. Six different swinepox viruses were constructed which expressed lac Z from EP1, LPl, LP2, EP1LP2, LP2EP2, or the SPV P01L promoter. Viruses were each used to infect A549, Chang liver, or 143B cells at 0.1 moi, and cells were rinsed between 2 and 3 hours later and then incubated for 4 days at 37 C. Each cell line maintained a different hierarchy of promoter activity, which was reproducible in following experiments. For example, the EP1, LP2EP2, and P01L promoters gave the most expression in 143B cells, while the LP2 was strongest in Chang liver cells, and the EP1LP2 in A549. In the Chang liver and A549 cells, expression form the P01L promoter was poorest, whereas in 143B, expression from LP2 was poorest. Therefore different human cell lines utilize pox promoters in dissimilar ways. This may reflect how far the swinepox virus can proceed along the replication pathway in different cell lines

These early and late promoters exhibited lower or higher lacZ activity depending on the human cell type infected by the recombinant swinepox virus . By choosing different promoters for different target tissues, one is able to regulate the amounts of foreign gene product delivered by the swinepox virus to target tissues.

Recombinant swinepox virus is useful as a vaccine for human infectious disease and to deliver therapeutic agents to humans. Recombinant swinepox virus is useful as a vaccine against viral or bacterial infection in humans, and as a therapeutic for cancer or genetic disease to deliver antibodies, tumor antigens, cell surface ligands and receptors, immune modulating molecules such as cytokines.

Example 40

S-SPV-003 Expression of lacZ in human cell lines Measurement of cytopathic effect and lacZ expression

When human cells are infected with SPV, a cytopathic effect is sometimes seen. In most cell lines, this cytopathic effect is evidenced by a change in the appearance of the cells, with cells becoming thinner and more ragged along the edges; cells look stressed. This phenomenon was assessed as follow:

indicates no difference between infected & uninfected cells;

+/- indicates that the monolayer is visibly different from uninfected, though most cells appear normal;

+ indicates that the monolayer is obviously affected, with most cells looking stressed. It should be noted that in certain cell lines (HeLa, CF500, 143B) , in which titers were obtained after serial passage, there was no evidence for replication of SPV, with one exception.

A549 was given a ++ for cytopathic effect in one instance, when cells appeared to round up and come off the plate during infection, though this observation was not repeated. A549 also showed evidence in another case of a ten-fold increase in titer during passage, suggesting that it might support limited viral replication.

B-galactosidase activity in A₂₆₀ units per cell lysate from 1/20 of a 35 mm dish: No Activity

+ 0.2-0.9 A₂₆₀ unit

++ 0.9-1,6 A₂ unit +++ greater than 1 . 6 A₂₆₀ units REFERENCES:

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(1991) . 37. T. Ben-Porat, et al . , Journal of Virology, volume 154, 325-334 (1986) . 38. F. Zuckerman, et al . , Vaccination and Control of Adjesky' s Disease, J. T. Van Oirchot (ed.) . Kluwer Academic Publishers, London, pp. 107-117 (1989) . 39. Paolette, et al . , Journal of Virology, volume 66, pp. 3424-3434 (June, 1992) .

40. M. W. Mellencamp, et al . , Journal of Clinical Microbiology, volume 27, pp. 2208-2213 (1989) .

41. L. A. Herzenberg, et al . , Selected Methods in Cellular Immunology, Freeman Publ . Co., San

Francisco, 351-372 (1980) .

42. Katz et al . , Journal of Virology 64, 1808-1811 (1990) .

43. Taniguchi, T. , et al . , Biochem. Biophys . Res . Commun. 115 1040-1047 (1983) .

44. Cochran, M.D. and Macdonald, R.D., WO 93/02104, published February 4, 1993.

45. Galibert, F., et al., Nature 281, 646-650 (1979) .

46. Thomsen, D.R., et al . , Proc . Na tl . Acad. Sci . USA 81, 659-663 (1984) .

47. Catalog Number 267402, Beckman Instruments, Inc., Fullerton California.

48. Whalley, J.M., et al., Journal of General Virology 57 307-323 (1981) . 49. Collett, M.S., et al . , Virology 165 200-208 (1988) .

50. Schodel, F. et al . , Journal of Virology 66, 106- 114 (1992) .

51. Cochran, M.D., WO 93/25665, published December 23, 1993.

52. CA. Hjerpe, The bovine Respiratory Disease Complex. Ed. by J.L. Howard, Philadelphia, W.B. Saunders Co., 670-680 (1986) .

53. F. Fenner, et al., Veterinary Virology. Academic Press, Inc., Orlando Florida, 183-202 (1987) .

54. A. Leutz, et al . , EMBO Journal 8: 175-182 (1989) . 55. M.J. Sekellick, et al . , Journal of Interferon Research 14: 71-79 (1994) .

56. S.J. Child, et al . , Virology 174: 625-629 (1990) .

57. G.P. Johnson, et al . Virology 196: 381-401 (1993) .

58. R.F Massung, et al . Virology 201: 215-240 (1994) .

60. Child, S.J. et al. , Virology 174, 625-629 (1990) .

61. T.R. Phillips, et al. , J. Virology 64, 4605-4613 (1990) 62. Massung, R.F. And Moyer, R.W., Virology 180, 347- 354 (1990) .

SEQUENCE LISTING

(1) GENERAL INFORMATION:

(l) APPLICANT: Cochran, Mark D. Junker, David E.

(n) TITLE OF INVENTION: Recombinant Swinepox Virus

(ill) NUMBER OF SEQUENCES: 231

(iv) CORRESPONDENCE ADDRESS:

(A) ADDRESSEE: John P. White

(B) STREET: 1185 Avenue of the Americas

(C) CITY: New York

(D) STATE: New York

(E) COUNTRY: USA

(F) ZIP: 10036

(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: Not Yet Known

(B) FILING DATE: 19-JAN-1996

(C) CLASSIFICATION:

(vni) ATTORNEY/AGENT INFORMATION: (A) NAME: White, John P (B) REGISTRATION NO: 28,678

(ix) TELECOMMUNICATION INFORMATION:

(A) TELEPHONE: (212) 278-0400

(B) TELEFAX: (212) 391-0525

(2) INFORMATION FOR SEQ ID NO:l:

(l) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 599 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(n) MOLECULE TYPE: DNA (genomic) (ill) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(vi) ORIGINAL SOURCE:

(A) ORGANISM: Swinepox virus

(B) STRAIN. Kasza

(C) INDIVIDUAL ISOLATE: S-SPV-001

(vn) 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:1:

AATGTATCCA GAGTTGTTGA ATGCCTTATC GTACCTAATA TTAATATAGA GTTATTAACT 60

GAATAAGTAT ATATAAATGA TTGTTTTTAT AATGTTTGTT ATCGCATTTA GTTTTGCTGT 120

ATGGTTATCA TATACATTTT TAAGGCCGTA TATGATAAAT GAAAATATAT AAGCACTTAT 180

TTTTGTTAGT ATAATAACAC A ATG CCG TCG TAT ATG TAT CCG AAG AAC GCA 231

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 279 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 327 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 375 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 423 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 471 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 519 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 567 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 599

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 Ile 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 He 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_start= 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 47

Asp He Lys Ser Cys Lys Cys Ser He Cys Ser Asp Ser He Thr 1 5 10 15 CAT CAT ATA TAT GAA ACA ACA TCA TGT ATA AAT TAT AAA TCT ACC GAT 95 His His He Tyr Glu Thr Thr Ser Cys He 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 143 Asn Asp Leu Met He 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 191 Gly Met He His Pro Asn Leu He 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 239 He Gly Leu Leu Thr Gly Asp He 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 287 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 335 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 383 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 431 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 479 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 527 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 575 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 623 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 672 Lys Val Ala Thr He He Leu Glu Glu He Lys Glu He 210 215 220

TTTCTTTTAA ATAAATAAAA ATACTTTTTT TTTTAAACAA GGGGTGCTAC CTTGTCTAAT 732

TGTATCTTGT ATTTTGGATC TGATGCAAGA TTATTAAATA ATCGTATGAA AAAGTAGTAG 792

ATATAGTTTA TATCGTTACT GGACATGATA TTATGTTTAG TTAATTCTTC TTTGGCATGA 852

ATTCTACACG TCGGANAAGG TAATGTATCT ATAATGGTAT AAAGCTT 899

(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: -23.2 (C) UNITS: %G

( i) 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 Ile 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:7:

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 60 TTGTCCGACG TGTAGAATTC ATGCCAAAGA AGAATTAACT AA 102

(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:

(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= ι

/standard_name= "Translation of synthetic DNA sequence"

(ix) FEATURE:

(A) NAME/KEY: CDS

(B) LOCATION: 100..102

(C) IDENTIFICATION METHOD: experimental

(D) OTHER INFORMATION: /partial

/codon_starts ioo /functions "marker enzyme" /products "Beta-Galactosidase" /evidences EXPERIMENTAL /gene= "lacZ" /numbers 2 /citations ( [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 60

TTTTTTTTTT TTTTTGGCAT ATAA ATG AAT TCG GAT CCC GTC 102

Met Asn Ser Asp Pro Val

1 5

(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: peptide

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 11:

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

(ii) MOLECULE TYPE: peptide

(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 /functions "marker enzyme" /product= "Beta-galactosidase" /evidences EXPERIMENTAL

/gene= "lacZ" /numbers ι /citations { [1] )

(ix) FEATURE:

(A) NAME/KEY: CDS

(B) LOCATION: 73..78

(C) IDENTIFICATION METHOD: experimental

(D) OTHER INFORMATION: /codon_start= 73

/functions "Translational finish of hybrid protein"

/products "C-terminal peptide"

/evidences EXPERIMENTAL

/numbers 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

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:13:

AGC CCG TCA GTA TCG GCG GAA ATC CAG CTG AGC GCC GGT CGC TAC CAT 48 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 98

Tyr Gin Leu Val Trp Cys Gin Lys Asp Pro 20 25

AAGAC 103

(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: peptide

(xi) 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: peptide

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 15:

Asp 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 48

(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 /functions "marker enzyme" /products "Beta-Galactosidase" /evidences EXPERIMENTAL /gene= "lacZ" /numbers 2 /citations ( [1] )

(ix) FEATURE:

(A) NAME/KEY: CDS

(B) LOCATION: 76..90

(D) OTHER INFORMATION: /partial /codon_start= 76

/functions "Translational start of hybrid protein" /products "N-terminal peptide" /numbers 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 60

TTTTCTATGC TATAA ATG AAT TCG GAT CCC GTC GTT TTA CAA 102

Met Asn Ser Asp Pro Val Val Leu Gin 1 5 (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: peptide

(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: peptide

(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_starts ι /functions "marker enzyme" /products "Beta-galactosidase" /evidences EXPERIMENTAL /numbers ι /citations ( [ι] )

(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) IDENTIFICATION METHOD: experimental

(D) OTHER INFORMATION: /codon_start= 177

/functions "Translational start of hybrid protein"

/products "N-terminal peptide"

/evidences EXPERIMENTAL

/standard_names "Translation of synthetic DNA sequence"

(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 Polycistronic 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

Ti mins, 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 48 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 99

Val Trp Cys Gin Lys Asp Pro 20

ACTCTAGAAA AAATTGAAAA ACTATTCTAA TTTATTGCAC GGAGATCTTT TTTTT TTTT 159 TTTTTTTTGG CATATAA ATG AAT TCG CTC GCA GCG CTA TTG GCG GCG 206

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: peptide

(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: peptide

(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: peptide

(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: peptide

(xi) 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) STRANDEDITOSS: 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) FEATURE:

(A) NAME/KEY: CDS

(B) LOCATION: 1..15

(D) OTHER INFORMATION: /partial /codon_star s ι /functions "glycoprotein" /products "PRV gp63" /genes "gp63" /numbers l /citations ( [1] )

(x) PUBLICATION INFORMATION:

(A) AUTHORS: Petrovskis, Erik A

Timmins, 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 55 Arg Val His His Glu 1 5

CCGGGTCGAC CTGCAGGCGG CCGGGTCGAC CTGCAGGCGG CCAGAC 101

(2) INFORMATION FOR SEQ ID NO: 7:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 5 amino acids

(B) TYPE: amino acid (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: peptide

(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 57

(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" /gene= "HN" /numbers ι

(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 Tyr 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 448 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 496 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 544 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 592 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 640 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 688 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 736 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 784 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 832 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 880 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 928 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 976 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 1024 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 1072 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 1120 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 1168 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 Lyε 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

Cys 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 57

(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_names "Translation of synthetic DNA sequence" ( ix) FEATURE :

(A) NAME/KEY: CDS

(B) LOCATION: 103..108

(C) IDENTIFICATION METHOD: experimental

(D) OTHER INFORMATION: /partial

/codon_starts 103

/products "NDV Heamagglutinin-Neuraminidase"

/evidences EXPERIMENTAL

/gene= ^»HN"

/numbers 2

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:32:

CATGTAGTCG ACTCTAGAAA AAATTGAAAA ACTATTCTAA TTTATTGCAC GGAGATCTTT 60 _{ττχτττττττ 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: peptide

(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 1

/standard_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_starts 85 /functions "marker enzyme" /products "Beta-galactosidase" /evidences EXPERIMENTAL /genes "lacZ" /numbers 2 /citations ( [l] )

(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 60

ATGCTATAA ATG AAT TCG GAT CCC GTC GTT TTA CAA CGT CGT GAC TGG 108

Met Asn Ser Asp Pro Val Val Leu Gin Arg Arg Asp Trp 1 5 10

(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: peptide

(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: peptide

(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_starts ι /functions "marker enzyme" /products "Beta-galactosidase" /evidences EXPERIMENTAL /gene= "lacZ" /numbers l /citations ( [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

/numbers 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 20

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: peptide

(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 57

(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 primer (xi) SEQUENCE DESCRIPTION: SEQ ID NO:41: CGCGAATTCG CTCGCAGCGC TATTGGC 27

(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 primer

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:42: GTAGGAGTGG CTGCTGAAG 19

(2) INFORMATION FOR SEQ ID NO:43:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 70 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

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:43: AAAAATTGAA AAACTATTCT AATTTATTGC ACGGAGATCT _TTTTTTTTTT TTTTTTTTTG 60 GCATATAAAT 70

(2) INFORMATION FOR SEQ ID NO:44:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 74 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

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:44: TTTTTTTTTT TTTTTTTTTT GGCATATAAA TAGATCTGTA TCCTAAAATT GAATTGTAAT 60 TATCGATAAT AAAT 74

(2) INFORMATION FOR SEQ ID NO:45:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 37 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

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:45: GTATCCTAAA ATTGAATTGT AATTATCGAT AATAAAT 37

(2) INFORMATION FOR SEQ ID NO:46:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 41 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

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:46: CGACTCTAGA ATTTCATTTT GTTTTTTTCT ATGCTATAAA T 41

(2) INFORMATION FOR SEQ ID NO:47:

(i) SEQUENCE CHARACTERISTICS :

(A) LENGTH: 60 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

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:47: CACATACGAT TTAGGTGACA CTATAGAATA CAAGCTTTGA GTCTATTGGT TATTTATACG 60

(2) INFORMATION FOR SEQ ID NO:48:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 123 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: 100..123

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:48:

TGAATATATA GCAAATAAAG GAAAAATTGT TATCGTTGCT GCATTAGATG GAACATAGGT 60

CGACTCTAGA ATTTCATTTT GTTTTTTTCT ATGCTATAA ATG AAT TCG GAT CCC 114

Met Asn Ser Asp Pro 1 5

GTC GTT TTA 123

Val Val Leu

(2) INFORMATION FOR SEQ ID NO:49:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 8 amino acids

(B) TYPE: amino acid (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: peptide

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:49:

Met Asn Ser Asp Pro Val Val Leu 1 5

(2) INFORMATION FOR SEQ ID NO:50:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 132 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: 1..63

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 50:

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 ACCTATGAAC GTAAACCATT 100

Gin Lys Asp Pro 20

TGGTAATATT CTTAATCTTA TACCATTATC GG 132

(2) INFORMATION FOR SEQ ID NO: 51:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 20 amino acids

(B) TYPE: amino acid (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 51:

Glu He Gin Leu Ser Ala Gly Arg Tyr His Tyr Gin Leu Val Trp Cys 1 5 10 15

Gin Lys Asp Pro 20

(2) INFORMATION FOR SEQ ID NO:52:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 66 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 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:52: TCTACTATTG TATATATAGG ATCCCCGGGC GAGCTCGAAT TCGTAATCAT GGTCATAGCT 60 GTTTCC 66

(2) INFORMATION FOR SEQ ID NO:53:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 51 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

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:53: ACAGGAAACA GCTATGACCA TGATTACGAA TTCGAGCTCG CCCGGGGATC T 51

(2) INFORMATION FOR SEQ ID NO:54:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 104 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 : 81 . . 104

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:54:

AAATATATAA ATACCATGTT AGAATTTGGT CTGCTGCAGG TCGACTCTAG AATTTCATTT 60

TGTTTTTTTC TATGCTATAA ATG AAT TCG GAT CCC GTC GTT TTA 104

Met Asn Ser Asp Pro Val Val Leu 1 5

(2) INFORMATION FOR SEQ ID NO:55:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 8 amino acids

(B) TYPE: amino acid (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: peptide

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:55:

Met Asn Ser Asp Pro Val Val Leu 1 5

(2) INFORMATION FOR SEQ ID NO: 56:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 150 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: 1..63

(i ) FEATURE:

(A) NAME/KEY: CDS

(B) LOCATION: 130..150

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:56:

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 ACCCGGTCGA CTCTAGAAAG ATCTGTATCC 100

Gin Lys Asp Pro 20

TAAAATTGAA TTGTAATTAT CGATAATAA ATG AAT TCC GGC ATG GCC TCG 150

Met Asn Ser Gly Met Ala Ser 1 5

(2) INFORMATION FOR SEQ ID NO:57:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 20 amino acids

(B) TYPE: amino acid (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: peptide

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 57:

Glu He Gin Leu Ser Ala Gly Arg Tyr His Tyr Gin Leu Val Trp Cys 1 5 10 15

Gin Lys Asp Pro 20

(2) INFORMATION FOR SEQ ID NO: 58:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 7 amino acids

(B) TYPE: amino acid (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: peptide

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:58:

Met Asn Ser Gly Met Ala Ser 1 5

(2) INFORMATION FOR SEQ ID NO:59:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 109 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 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:59: CCATGCTCTA GAGGATCCCC GGGCGAGCTC GAATTCGGAT CCATAATTAA TTAATTAATT 60 TTTATCCCGG GTCGACCGGG TCGACCTGCA GCCTACATGG AAATCTACC 109

(2) INFORMATION FOR SEQ ID NO:60:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 51 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

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:60: TAATGTATCT ATAATGGTAT AAAGCTTGTA TTCTATAGTG TCACCTAAAT C 51

(2) INFORMATION FOR SEQ ID NO:61:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 51 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

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:61: ACAGGAAACA GCTATGACCA TGATTACGAA TTCGAGCTCG CCCGGGGATC T 51

(2) INFORMATION FOR SEQ ID NO:62:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 104 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: 81..104

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:62:

AAATATATAA ATACCATGTT AGAATTTGGT CTGCTGCAGG TCGACTCTAG AATTTCATTT 60

TGTTTTTTTC TATGCTATAA ATG AAT TCG GAT CCC GTC GTT TTA 104

Met Asn Ser Asp Pro Val Val Leu 1 5

(2) INFORMATION FOR SEQ ID NO:63:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 8 amino acids

(B) TYPE: amino acid (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: peptide

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:63:

Met Asn Ser Asp Pro Val Val Leu

1 5

(2) INFORMATION FOR SEQ ID NO:64:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 182 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: 1..63

(ix) FEATURE:

(A) NAME/KEY: CDS

(B) LOCATION: 156..182

(Xi) SEQUENCE DESCRIPTION: SEQ ID NO:64:

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 ACCCGGTCGA CTCTAGAAAA AATTGAAAAA 100

Gin Lys Asp Pro 20

CTATTCTAAT TTATTGCACG GAGATCTTTT TTTTTTTTTT TTTTTTGGCA TATAA ATG 158

Met

1

AAT TCC GGC ATG GCC TCG CTC GCG 182

Asn Ser Gly Met Ala Ser Leu Ala 5

(2) INFORMATION FOR SEQ ID NO:65:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 20 amino acids

(B) TYPE: amino acid (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: peptide

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:65:

Glu He Gin Leu Ser Ala Gly Arg Tyr His Tyr Gin Leu Val Trp Cys 1 5 10 15

Gin Lys Asp Pro 20

(2) INFORMATION FOR SEQ ID NO:66:

(i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 9 amino acids

(B) TYPE: amino acid (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: peptide

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:66:

Met Asn Ser Gly Met Ala Ser Leu Ala 1 5

(2) INFORMATION FOR SEQ ID NO:67:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 109 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

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:67: CCATGCTCTA GAGGATCCCC GGGCGAGCTC GAATTCGGAT CCATAATTAA TTAATTAATT 60 TTTATCCCGG GTCGACCGGG TCGACCTGCA GCCTACATGG AAATCTACC 109

(2) INFORMATION FOR SEQ ID NO:68:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 51 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

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:68: TAATGTATCT ATAATGGTAT AAAGCTTGTA TTCTATAGTG TCACCTAAAT C 51

(2) INFORMATION FOR SEQ ID NO:69:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 51 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

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:69: ACAGGAAACA GCTATGACCA TGATTACGAA TTCGAGCTCG CCCGGGGATC T 51

(2) INFORMATION FOR SEQ ID NO: 70:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 104 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: 81..104

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:70:

AAATATATAA ATACCATGTT AGAATTTGGT CTGCTGCAGG TCGACTCTAG AATTTCATTT 60

TGTTTTTTTC TATGCTATAA ATG AAT TCG GAT CCC GTC GTT TTA 104

Met Asn Ser Asp Pro Val Val Leu 1 5

(2) INFORMATION FOR SEQ ID NO:71:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 8 amino acids

(B) TYPE: amino acid (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: peptide

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:71:

Met Asn Ser Asp Pro Val Val Leu 1 5

(2) INFORMATION FOR SEQ ID NO:72:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 180 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: 1..63

(ix) FEATURE:

(A) NAME/KEY: CDS (B) LOCATION: 160..180

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:72: 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 ACCCGGTCGA CTCTAGATTT TTTTTTTTTT 100

Gin Lys Asp Pro 20

TTTTTTTGGC ATATAAATAG ATCTGTATCC TAAAATTGAA TTGTAATTAT CGATAATAA 159

ATG AAT TCC GGC ATG GCC TCG 180

Met Asn Ser Gly Met Ala Ser 1 5

(2) INFORMATION FOR SEQ ID NO:73:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 20 amino acids

(B) TYPE: amino acid (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: peptide

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:73:

Glu He Gin Leu Ser Ala Gly Arg Tyr His Tyr Gin Leu Val Trp Cys 1 5 10 15

Gin Lys Asp Pro 20

(2) INFORMATION FOR SEQ ID NO: 74 :

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 7 amino acids

(B) TYPE: amino acid (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: peptide

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 74

Met Asn Ser Gly Met Ala Ser 1 5

(2) INFORMATION FOR SEQ ID NO:75:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 109 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

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:75: CCATGCTCTA GAGGATCCCC GGGCGAGCTC GAATTCGGAT CCATAATTAA TTAATTAATT 60 TTTATCCCGG GTCGACCGGG TCGACCTGCA GCCTACATGG AAATCTACC 109

(2) INFORMATION FOR SEQ ID NO: 76:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 51 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

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 76: TAATGTATCT ATAATGGTAT AAAGCTTGTA TTCTATAGTG TCACCTAAAT C 51

(2) INFORMATION FOR SEQ ID NO: 77:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 51 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

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 77: ACAGGAAACA GCTATGACCA TGATTACGAA TTCGAGCTCG CCCGGGGATC T 51

(2) INFORMATION FOR SEQ ID NO: 78:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 117 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: 94..117

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 78:

GGTCTGCTGC AGGTCGACTC TAGAAAAAAT TGAAAAACTA TTCTAATTTA TTGCACGGAG 60

ATCTTTTTTT TTTTTTTTTT TTTTGGCATA TAA ATG AAT TCC GGC TTC AGT AAC ATA 117

Met Asn Ser Gly Phe Ser Asn He 1 5 8

(2) INFORMATION FOR SEQ ID NO:79:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 8 amino acids

(B) TYPE: amino acid (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: peptide

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 79:

Met Asn Ser Gly Phe Ser Asn He 1 5

(2) INFORMATION FOR SEQ ID NO:80: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 126 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: 103..126

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 80:

CGCAACATAC CTAACTGCTT CATTTCTGAT CCATAATTAA TTAATTTTTA TCCCGGCGCG 60

CCTCGACTCT AGAATTTCAT TTTGTTTTTT TCTATGCTAT AA ATG AAT TCG GAT 114

Met Asn Ser Asp

1

CCC GTC GTT TTA 126

Pro Val Val Leu 5

(2) INFORMATION FOR SEQ ID NO:81:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 8 amino acids

(B) TYPE: amino acid (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: peptide

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:81:

Met Asn Ser Asp Pro Val Val Leu 1 5

(2) INFORMATION FOR SEQ ID NO: 82:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 96 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: 1..63

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 82:

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 ACCTGCAGCC TACATG 96

Gin Lys Asp Pro 20

(2) INFORMATION FOR SEQ ID NO:83:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 20 amino acids

(B) TYPE: amino acid (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: peptide

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:83:

Glu He Gin Leu Ser Ala Gly Arg Tyr His Tyr Gin Leu Val Trp Cys 1 5 10 15

Gin Lys Asp Pro 20

(2) INFORMATION FOR SEQ ID NO: 84:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 51 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

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:84: TAATGTATCT ATAATGGTAT AAAGCTTGTA TTCTATAGTG TCACCTAAAT C 51

(2) INFORMATION FOR SEQ ID NO:85:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 51 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

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:85: ACAGGAAACA GCTATGACCA TGATTACGAA TTCGAGCTCG CCCGGGGATC T 51

(2) INFORMATION FOR SEQ ID NO:86:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 124 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: 104..124

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:86:

GTATAGCGGC CGCCTGCAGG TCGACTCTAG ATTTTTTTTT TTTTTTTTTT TGGCATATAA 60

ATAGATCTGT ATCCTAAAAT TGAATTGTAA TTATCGATAA TAA ATG AAT TCG CTA CTT 113

Met Asn Ser Leu Leu 1 5

GGA ACT 124

Gly Thr

(2) INFORMATION FOR SEQ ID NO:87:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 7 amino acids

(B) TYPE: amino acid (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: peptide

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:87:

Met Asn Ser Leu Leu Gly Thr 1 5

(2) INFORMATION FOR SEQ ID NO:88:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 126 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: 1..12

(ix) FEATURE:

(A) NAME/KEY: CDS

(B) LOCATION: 103..126 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:88:

ATA AAA ATG TGATTAAGTC TGAATGTGGA TCCATAATTA ATTAATTTTT 49

He Lys Met

1

ATCCCGGCGC GCCTCGACTC TAGAATTTCA TTTTGTTTTT TTCTATGCTA TAA ATG 105

Met

1

AAT TCG GAT CCC GTC GTT TTA 126

Asn Ser Asp Pro Val Val Leu 5

(2) INFORMATION FOR SEQ ID NO:89:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 3 amino acids

(B) TYPE: amino acid (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: peptide

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:89:

He Lys Met

1

(2) INFORMATION FOR SEQ ID NO:90:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 8 amino acids

(B) TYPE: amino acid (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: peptide

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 90:

Met Asn Ser Asp Pro Val Val Leu 1 5

(2) INFORMATION FOR SEQ ID NO: 91:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 116 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: 1..63

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 91:

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 AGGCGCGCCG GGTCGACCTG 100

Gin Lys Asp Pro 20

CAGGCGGCCG CTATAC 116

(2) INFORMATION FOR SEQ ID NO:92:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 20 amino acids

(B) TYPE: amino acid (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: peptide

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 92:

Glu He Gin Leu Ser Ala Gly Arg Tyr His Tyr Gin Leu Val Trp Cys 1 5 10 15

Gin Lys Asp Pro 20

(2) INFORMATION FOR SEQ ID NO: 93:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 51 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

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:93: TAATGTATCT ATAATGGTAT AAAGCTTGTA TTCTATAGTG TCACCTAAAT C 51

(2) INFORMATION FOR SEQ ID NO:94:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 51 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

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 94: ACAGGAAACA GCTATGACCA TGATTACGAA TTCGAGCTCG CCCGGGGATC T 51

(2) INFORMATION FOR SEQ ID NO: 95:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 124 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: 104..124

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:95: GTATAGCGGC CGCCTGCAGG TCGACTCTAG ATTTTTTTTT TTTTTTTTTT TGGCATATAA 60 ATAGATCTGT ATCCTAAAAT TGAATTGTAA TTATCGATAA TAA ATG AAT TCC CCT GCC 113

Met Asn Ser Pro Ala 1 5

GCC CGG 124

Ala Arg

(2) INFORMATION FOR SEQ ID NO:96:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 7 amino acids

(B) TYPE: amino acid (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: peptide

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 96:

Met Asn Ser Pro Ala Ala Arg

1 5

(2) INFORMATION FOR SEQ ID NO:97:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 126 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: 1..36

(ix) FEATURE:

(A) NAME/KEY: CDS

(B) LOCATION: 103..126

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:97:

CTC CAG GAG CCC GCT CGC CTC GAG CGG GAT CCA TAATTAATTA ATTTTTATCC 53 Leu Gin Glu Pro Ala Arg Leu Glu Arg Asp Pro 1 5 10 CGGCGCGCCT CGACTCTAGA ATTTCATTTT GTTTTTTTCT ATGCTATAA ATG AAT 108

Met Asn

1

TCG GAT CCC GTC GTT TTA 126

Ser Asp Pro Val Val Leu 5

(2) INFORMATION FOR SEQ ID NO:98:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 11 amino acids

(B) TYPE: amino acid (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: peptide

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 98:

Leu Gin Glu Pro Ala Arg Leu Glu Arg Asp Pro 1 5 10

(2) INFORMATION FOR SEQ ID NO:99:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 8 amino acids

(B) TYPE: amino acid (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: peptide

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:99:

Met Asn Ser Asp Pro Val Val Leu 1 5

(2) INFORMATION FOR SEQ ID NO: 100:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 116 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: 1..63

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 100:

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 AGGCGCGCCG GGTCGACCTG 100

Gin Lys Asp Pro 20

CAGGCGGCCG CTATAC 116

(2) INFORMATION FOR SEQ ID NO: 101:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 20 amino acids

(B) TYPE: amino acid (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: peptide

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:101:

Glu He Gin Leu Ser Ala Gly Arg Tyr His Tyr Gin Leu Val Trp Cys 1 5 10 15

Gin Lys Asp Pro 20

(2) INFORMATION FOR SEQ ID NO:102:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 51 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

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:102: TAATGTATCT ATAATGGTAT AAAGCTTGTA TTCTATAGTG TCACCTAAAT C 51 (2) INFORMATION FOR SEQ ID NO:103:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 30 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

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:103: CCGAATTCCG GCTTCAGTAA CATAGGATCG 30

(2) INFORMATION FOR SEQ ID NO:104:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 20 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

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:104: GTACCCATAC TGGTCGTGGC 20

(2) INFORMATION FOR SEQ ID NO:105:

(i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 26 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

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 105: CCGGAATTCG CTACTTGGAA CTCTGG 26

(2) INFORMATION FOR SEQ ID NO: 106:

(i) SEQUENCE CHARACTERISTICS :

(A) LENGTH: 20 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

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:106: CATTGTCCCG AGACGGACAG 20

(2) INFORMATION FOR SEQ ID NO: 107:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 19 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

( ii) IMMEDIATE SOURCE:

(B) CLONE: 515-85.1

(viii) POSITION IN GENOME:

(B) MAP POSITION: -23.2

(C) UNITS: %G

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:107: CGCGATCCAA CTATCGGTG 19

(2) INFORMATION FOR SEQ ID NO:108:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 26 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

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:108: GCGGATCCAC ATTCAGACTT AATCAC 26

(2) INFORMATION FOR SEQ ID NO:109:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 30 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

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:109: ATGAATTCCC CTGCCGCCCG GACCGGCACC 30

(2) INFORMATION FOR SEQ ID NO:110:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 30 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

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:110: CATGGATCCC GCTCGAGGCG AGCGGGCTCC 30

(2) INFORMATION FOR SEQ ID NO:111:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 42 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

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:111: CTGGTTCGGC CCAGAATTCT ATGGGTCTCG CGCGGCTCGT GG 42

(2) INFORMATION FOR SEQ ID NO:112:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 42 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

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 112: CTCGCTCGCC CAGGATCCCT AGCGGAGGAT GGACTTGAGT CG 42

(2) INFORMATION FOR SEQ ID NO: 113:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 3628 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: 57..1226

(ix) FEATURE:

(A) NAME/KEY: CDS

(B) LOCATION: 1362..3395

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:113:

TTGAAGATGA ATGCATAGAG GAAGATGATG TCGANACGTC ATTATTTAAT GTATAAATGG 60

ATAAATTGTA TGCGGCAATA TTCGGCGTTT TTATGACATC TAAAGATGAT GATTTTAATA 120

ACTTTATAGA AGTTGTAAAA TCTGTATTAA CAGATACATC ANCTAATCAT ACAATATCGT 180

CGTCCAATAA TAATACATGG ATATATATAT TTCTAGCGAT ATTATTTGGT GTTATGGNAT 240

TATTAGTTTT TANTTTGTAT GTAGAAGTTC CTAAACCNAC TTANATGGAG GAAGCAGATA 300

ACCNACTCGT TNTAAATAGT ATTAGTGCTA GAGCATTGGN GGCATTTTTT GTATCTAAAA 360

NTANTGATAT GGTCGNTGAA NTAGTTNCCC AAAAATNTCC NCCAAAGAAG ANATCACAAA 420

TAAAACGCAT AGATACACGA ATTCCTATTG ATCTTATTAA TCAACAATTC GTTAAAAGAT 480

TTAAACTAGA AAATTATAAA AATGGAATTT TATCCGTTCT TATCAATAGT TTAGTCGAAA 540

ATAATTACTT TGAACAAGAT GGTAAACTTA ATAGCAGTGA TATTGATGAA TTAGTGCTCA 600

CAGACATAGA GAAAAAGATT TTATCGTTGA TTCCTAGATG TTCTCCTCTT TATATAGATA 660

TCAGTGACGT TAAAGTTCTC GCATCTAGGT TAANNAAAAG TGCTAAATCA TTTACGTTTA 720

ATGATCATGA ATATATTATA CAATCTGATA AAATAGAGGA ATTAATAAAT AGTTTATCTA 780

GAAACCATGA TATTATACTA GATGAAAAAA GTTCTATTAA AGACAGCATA TATATACTAT 840

CTGATGATCT TTTGAATATA CTTCGTGAAA GATTATTTAG ATGTCCACAG GTTAAAGATA 900

ATACTATTTC TAGAACACGT CTATATGATT ATTTTACTAG AGTGTCAAAG AAAGAAGAAG 960

CGAAAATATA CGTTATATTG AAAGATTTAA AGATTGCTGA TATACTCGGT ATCGAAACAG 1020

TAACGATAGG ATCATTTGTA TATACGAAAT ATAGCATGTT GATTAATTCA ATTTCGTCTA 1080

ATGTTGATAG ATATTCAAAA AGGTTCCATG ACTCTTTTTA TGAAGATATT GCGGAATTTA 1140

TAAAGGATAA TGAAAAAATT AATGTATCCA GAGTTGTTGA ATGCCTTATC GTACCTAATA 1200

TTAATATAGA GTTATTAACT GAATAAGTAT ATATAAATGA TTGTTTTTAT AATGTTTGTT 1260

ATCGCATTTA GTTTTGCTGT ATGGTTATCA TATACATTTT TAAGGCCGTA TATGATAAAT 1320

GAAAATATAT AAGCACTTAT TTTTGTTAGT ATAATAACAC AATGCCGTCG TATATGTATC 1380

CGAAGAACGC AAGAAAAGTA ATTTCAAAGA TTATATCATT ACAACTTGAT ATTAAAAAAC 1440

TTCCTAAAAA ATATATAAAT ACCATGTTAG AATTTGGTCT ACATGGAAAT CTACCAGCTT 1500

GTATGTATAA AGATGCCGTA TCATATGATA TAAATAATAT AAGATTTTTA CCTTATAATT 1560 GTGTTATGGT TAAAGATTTA ATAAATGTTA TAAAATCATC ATCTGTAATA GATACTAGAT 1620

TACATCAATC TGTATTAAAA CATCGTAGAG CGTTAATAGA TTACGGCGAT CAAGACATTA 1680

TCACTTTAAT GATCATTAAT AAGTTACTAT CGATAGATGA TATATCCTAT ATATTAGATA 1740

AAAAAATAAT TCATGTAACA AAAATATTAA AAATAGACCC TACAGTAGCC AATTCAAACA 1800

TGAAACTGAA TAAGATAGAG CTTGTAGATG TAATAACATC AATACCTAAG TCTTCCTATA 1860

CATATTTATA TAATAATATG ATCATTGATC TCGATACATT ATTATATTTA TCCGATGCAT 1920

TCCACATACC CCCCACACAT ATATCATTAC GTTCACTTAG AGATATAAAC AGGATTATTG 1980

AATTGCTTAA AAAATATCCG AATAATAATA TTATTGATTA TATATCCGAT AGCATAAAAT 2040

CAAATAGTTC ATTCATTCAC ATACTTCATA TGATAATATC AAATATGTTT CCTGCTATAA 2100

TCCCTAGTGT AAACGATTTT ATATCTACCG TAGTTGATAA AGATCGACTT ATTAATATGT 2160

ATGGGATTAA GTGTGTTGCT ATGTTTTCGT ACGATATAAA CATGATCGAT TTAGAGTCAT 2220

TAGATGACTC AGATTACATA TTTATAGAAA AAAATATATC TATATACGAC GTTAAATGTA 2280

GAGATTTTGC GAATATGATT AGAGATAAGG TTAAAAGAGA AAAGAATAGA ATATTAACTA 2340

CGAAATGTGA AGATATTATA AGATATATAA AATTATTCAG TAAAAATAGA ATAAACGATG 2400

AAAATAATAA GGTGGAGGAG GTGTTGATAC ATATTGATAA TGTATCTAAA AATAATAAAT 2460

TATCACTGTC TGATATATCA TCTTTAATGG ATCAATTTCG TTTAAATCCA TGTACCATAA 2520

GAAATATATT ATTATCTTCA GCAACTATAA AATCAAAACT ATTAGCGTTA CGGGCAGTAA 2580

AAAACTGGAA ATGTTATTCA TTGACAAATG TATCAATGTA TAAAAAAATA AAGGGTGTTA 2640

TCGTAATGGA TATGGTTGAT TATATATCTA CTAACATTCT TAAATACCAT AAACAATTAT 2700

ATGATAAAAT GAGTACGTTT GAATATAAAC GAGATATTAA ATCATGTAAA TGCTCGATAT 2760

GTTCCGACTC TATAACACAT CATATATATG AAACAACATC ATGTATAAAT TATAAATCTA 2820

CCGATAATGA TCTTATGATA GTATTGTTCA ATCTAACTAG ATATTTAATG CATGGGATGA 2880

TACATCCTAA TCTTATAAGC GTAAAAGGAT GGGGTCCCCT TATTGGATTA TTAACGGGTG 2940

ATATAGGTAT TAATTTAAAA CTATATTCCA CCATGAATAT AAATGGGCTA CGGTATGGAG 3000

ATATTACGTT ATCTTCATAC GATATGAGTA ATAAATTAGT CTCTATTATT AATACACCCA 3060

TATATGAGTT AATACCGTTT ACTACATGTT GTTCACTCAA TGAATATTAT TCAAAAATTG 3120

TGATTTTAAT AAATGTTATT TTAGAATATA TGATATCTAT TATATTATAT AGAATATTGA 3180

TCGTAAAAAG ATTTAATAAC ATTAAAGAAT TTATTTCAAA AGTCGTAAAT ACTGTACTAG 3240

AATCATCAGG CATATATTTT TGTCAGATGC GTGTACATGA ACAAATTGAA TTGGAAATAG 3300

ATGAGCTCAT TATTAATGGA TCTATGCCTG TACAGCTTAT GCATTTACTT CTAAAGGTAG 3360

CTACCATAAT ATTAGAGGAA ATCAAAGAAA TATAACGTAT TTTTTCTTTT AAATAAATAA 3420

AAATACTTTT TTTTTTAAAC AAGGGGTGCT ACCTTGTCTA ATTGTATCTT GTATTTTGGA 3480

TCTGATGCAA GATTATTAAA TAATCGTATG AAAAAGTAGT AGATATAGTT TATATCGTTA 3540

CTGGACATGA TATTATGTTT AGTTAATTCT TCTTTGGCAT GAATTCTACA CGTCGGANAA 3600 GGTAATGTAT CTATAATGGT ATAAAGCT 3628

(2) INFORMATION FOR SEQ ID NO:114:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 389 amino acids

(B) TYPE: amino 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

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:114:

Met Asp Lys Leu Tyr Ala Ala He Phe Gly Val Phe Met Thr Ser Lys 1 5 10 15

Asp Asp Asp Phe Asn Asn Phe He Glu Val Val Lys Ser Val Leu Thr 20 25 30

Asp Thr Ser Xaa Asn His Thr He Ser Ser Ser Asn Asn Asn Thr Trp 35 40 45

He Tyr He Phe Leu Ala He Leu Phe Gly Val Met Xaa Leu Leu Val 50 55 60

Phe Xaa Leu Tyr Val Glu Val Pro Lys Pro Thr Xaa Met Glu Glu Ala 65 70 75 80

Asp Asn Xaa Leu Val Xaa Asn Ser He Ser Ala Arg Ala Leu Xaa Ala 85 90 95

Phe Phe Val Ser Lys Xaa Xaa Asp Met Val Xaa Glu Xaa Val Xaa Gin 100 105 110

Lys Xaa Pro Pro Lys Lys Xaa Ser Gin He Lys Arg He Asp Thr Arg 115 120 125

He Pro He Asp Leu He Asn Gin Gin Phe Val Lys Arg Phe Lys Leu 130 135 140

Glu Asn Tyr Lys Asn Gly He Leu Ser Val Leu He Asn Ser Leu Val 145 150 155 160

Glu Asn Asn Tyr Phe Glu Gin Asp Gly Lys Leu Asn Ser Ser Asp He 165 170 175

Asp Glu Leu Val Leu Thr Asp He Glu Lys Lys He Leu Ser Leu He 180 185 190 Pro Arg Cys Ser Pro Leu Tyr He Asp He Ser Asp Val Lys Val Leu 195 200 205

Ala Ser Arg Leu Xaa Lys Ser Ala Lys Ser Phe Thr Phe Asn Asp His 210 215 220

Glu Tyr He He Gin Ser Asp Lys He Glu Glu Leu He Asn Ser Leu 225 230 235 240

Ser Arg Asn His Asp He He Leu Asp Glu Lys Ser Ser He Lys Asp 245 250 255

Ser He Tyr He Leu Ser Asp Asp Leu Leu Asn He Leu Arg Glu Arg 260 265 270

Leu Phe Arg Cys Pro Gin Val Lys Asp Asn Thr He Ser Arg Thr Arg 275 280 285

Leu Tyr Asp Tyr Phe Thr Arg Val Ser Lys Lys Glu Glu Ala Lys He 290 295 300

Tyr Val He Leu Lys Asp Leu Lys He Ala Asp He Leu Gly He Glu 305 310 315 320

Thr Val Thr He Gly Ser Phe Val Tyr Thr Lys Tyr Ser Met Leu He 325 330 335

Asn Ser He Ser Ser Asn Val Asp Arg Tyr Ser Lys Arg Phe His Asp 340 345 350

Ser Phe Tyr Glu Asp He Ala Glu Phe He Lys Asp Asn Glu Lys He 355 360 365

Asn Val Ser Arg Val Val Glu Cys Leu He Val Pro Asn He Asn He 370 375 380

Glu Leu Leu Thr Glu 385

(2) INFORMATION FOR SEQ ID NO:115:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 677 amino acids

(B) TYPE: amino 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 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 115:

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 Thr Lys He Leu Lys He Asp Pro Thr Val Ala Asn 130 135 140

Ser Asn Met Lys Leu Asn Lys He Glu Leu Val Asp Val He Thr Ser 145 150 155 160

He Pro Lys Ser Ser Tyr Thr Tyr Leu Tyr Asn Asn Met He He Asp 165 170 175

Leu Asp Thr Leu Leu Tyr Leu Ser Asp Ala Phe His He Pro Pro Thr 180 185 190

His He Ser Leu Arg Ser Leu Arg Asp He Asn Arg He He Glu Leu 195 200 205

Leu Lys Lys Tyr Pro Asn Asn Asn He He Asp Tyr He Ser Asp Ser 210 215 220

He Lys Ser Asn Ser Ser Phe He His He Leu His Met He He Ser 225 230 235 240

Asn Met Phe Pro Ala He He Pro Ser Val Asn Asp Phe He Ser Thr 245 250 255

Val Val Asp Lys Asp Arg Leu He Asn Met Tyr Gly He Lys Cys Val 260 265 270

Ala Met Phe Ser Tyr Asp He Asn Met He Asp Leu Glu Ser Leu Asp 275 280 285

Asp Ser Asp Tyr He Phe He Glu Lys Asn He Ser He Tyr Asp Val 290 295 300

Lys Cys Arg Asp Phe Ala Asn Met He Arg Asp Lys Val Lys Arg Glu 305 310 315 320

Lys Asn Arg He Leu Thr Thr Lys Cys Glu Asp He He Arg Tyr He 325 330 335

Lys Leu Phe Ser Lys Asn Arg He Asn Asp Glu Asn Asn Lys Val Glu 340 345 350 Glu Val Leu He His He Asp Asn Val Ser Lys Asn Asn Lys Leu Ser 355 360 365

Leu Ser Asp He Ser Ser Leu Met Asp Gin Phe Arg Leu Asn Pro Cys 370 375 380

Thr He Arg Asn He Leu Leu Ser Ser Ala Thr He Lys Ser Lys Leu 385 390 395 400

Leu Ala Leu Arg Ala Val Lys Asn Trp Lys Cys Tyr Ser Leu Thr Asn 405 410 415

Val Ser Met Tyr Lys Lys He Lys Gly Val He Val Met Asp Met Val 420 425 430

Asp Tyr He Ser Thr Asn He Leu Lys Tyr His Lys Gin Leu Tyr Asp 435 440 445

Lys Met Ser Thr Phe Glu Tyr Lys Arg Asp He Lys Ser Cys Lys Cys 450 455 460

Ser He Cys Ser Asp Ser He Thr His His He Tyr Glu Thr Thr Ser 465 470 475 480

Cys He Asn Tyr Lys Ser Thr Asp Asn Asp Leu Met He Val Leu Phe 485 490 495

Asn Leu Thr Arg Tyr Leu Met His Gly Met He His Pro Asn Leu He 500 505 510

Ser Val Lys Gly Trp Gly Pro Leu He Gly Leu Leu Thr Gly Asp He 515 520 525

Gly He Asn Leu Lys Leu Tyr Ser Thr Met Asn He Asn Gly Leu Arg 530 535 540

Tyr Gly Asp He Thr Leu Ser Ser Tyr Asp Met Ser Asn Lys Leu Val 545 550 555 560

Ser He He Asn Thr Pro He Tyr Glu Leu He Pro Phe Thr Thr Cys 565 570 575

Cys Ser Leu Asn Glu Tyr Tyr Ser Lys He Val He Leu He Asn Val 580 585 590

He Leu Glu Tyr Met He Ser He He Leu Tyr Arg He Leu He Val 595 600 605

Lys Arg Phe Asn Asn He Lys Glu Phe He Ser Lys Val Val Asn Thr 610 615 620

Val Leu Glu Ser Ser Gly He Tyr Phe Cys Gin Met Arg Val His Glu 625 630 635 640

Gin He Glu Leu Glu He Asp Glu Leu He He Asn Gly Ser Met Pro 645 650 655

Val Gin Leu Met His Leu Leu Leu Lys Val Ala Thr He He Leu Glu 660 665 670

Glu He Lys Glu He 675

(2) INFORMATION FOR SEQ ID NO:116: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 43 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: Infectious bovine rhinotracheitis virus

(B) STRAIN: Cooper Strain

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:116: CTGGTTCGGC CCAGAATTCG ATGCAACCCA CCGCGCCGCC CCG 43

(2) INFORMATION FOR SEQ ID NO: 117:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 42 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: Infectious bovine rhinotracheitis virus

(B) STRAIN: Cooper Strain

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:117: CTCGCTCGCC CAGGATCCCT AGCGGAGGAT GGACTTGAGT CG 42

(2) INFORMATION FOR SEQ ID NO:118:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 31 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: Equine Influenza A neuraminidase

(B) STRAIN: Prague/56

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:118: GGGATCCATG AATCCTAATC AAAAACTCTT T 31 (2) INFORMATION FOR SEQ ID NO:119:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 31 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: Equine Influenza A neuraminidase

(B) STRAIN: Prague/56

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:119: GGGATCCTTA CGAAAAGTAT TTAATTTGTG C 31

(2) INFORMATION FOR SEQ ID NO: 120:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 42 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: Equine influenza A hemagglutinin

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:120:

GGAGGCCTTC ATGACAGACA ACCATTATTT TGATACTACT GA 42

(2) INFORMATION FOR SEQ ID NO: 121:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 40 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: Equine influenza A hemagglutinin

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:121:

GAAGGCCTTC TCAAATGCAA ATGTTGCATC TGATGTTGCC 40 (2) INFORMATION FOR SEQ ID NO:122:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 32 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: Equine Influenza A hemagglutinin

(B) STRAIN: Prague/56

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:122: GGGATCCATG AACACTCAAA TTCTAATATT AG 32

(2) INFORMATION FOR SEQ ID NO: 123:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 30 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: Equine Influenza A hemagglutinin

(B) STRAIN: Prague/56

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:123: GGGATCCTTA TATACAAATA GTGCACCGCA 30

(2) INFORMATION FOR SEQ ID NO: 124:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 30 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: Equine Influenza A neuraminidase

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:124: GGGTCGACAT GAATCCAAAT CAAAAGATAA 30

(2) INFORMATION FOR SEQ ID NO:125:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 29 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: Equine Influenza A neuraminidase

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:125: GGGTCGACTT ACATCTTATC GATGTCAAA 29

(2) INFORMATION FOR SEQ ID NO: 126:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 33 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: Human

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:126:

CTCGAATTCG AAGTGGGCAA CGTGGATCCT CGC 33

(2) INFORMATION FOR SEQ ID NO:127:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 24 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: Human

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:127: CAGTTAGCCT CCCCCATCTC CCCA 24

(2) INFORMATION FOR SEQ ID NO:128:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 21 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: Equine herpesvirus type 1

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:128: CGGAATTCCT CTGGTTGCCG T 21

(2) INFORMATION FOR SEQ ID NO:129:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 22 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: Equine herpesvirus type 1

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 129:

GACGGTGGAT CCGGTAGGCG GT 22

(2) INFORMATION FOR SEQ ID NO:130:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 34 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: Bovine parainfluenza-3 virus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:130: TTATGGATCC TGCTGCTGTG TTGAACAACT TTGT 34

(2) INFORMATION FOR SEQ ID NO:131:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 38 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: Bovine parainfluenza-3 virus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:131: CCGCGGATCC CATGACCATC ACAACCATAA TCATAGCC 38

(2) INFORMATION FOR SEQ ID NO:132:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 43 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: Bovine parainfluenza-3 virus

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:132: CGTCGGATCC CTTAGCTGCA GTTTTTTGGA ACTTCTGTTT TGA 43

(2) INFORMATION FOR SEQ ID NO: 133:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 40 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: Bovine parainfluenza-3 virus (xi) SEQUENCE DESCRIPTION: SEQ ID NO:133: CATAGGATCC CATGGAATAT TGGAAACACA CAAACAGCAC 40

(2) INFORMATION FOR SEQ ID NO:134:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 42 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: Bovine viral diarrhea virus

(B) STRAIN: Singer Strain

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 134: ACGTCGGATC CCTTACCAAA CCACGTCTTA CTCTTGTTTT CC 42

(2) INFORMATION FOR SEQ ID NO:135:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 40 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: Bovine viral diarrhea virus

(B) STRAIN: Singer Strain

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:135: ACATAGGATC CCATGGGAGA AAACATAACA CAGTGGAACC 40

(2) INFORMATION FOR SEQ ID NO:136:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 33 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: Bovine viral diarrhea virus

(B) STRAIN: Singer Strain

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:136: CGTGGATCCT CAATTACAAG AGGTATCGTC TAC 33

(2) INFORMATION FOR SEQ ID NO:137:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 31 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: Bovine viral diarrhea virus

(B) STRAIN: Singer Strain

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:137: CATAGATCTT GTGGTGCTGT CCGACTTCGC A 31

(2) INFORMATION FOR SEQ ID NO:138:

(i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 37 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: Bovine respiratory syncytial virus

(B) STRAIN: Strain 375

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:138: TGCAGGATCC TCATTTACTA AAGGAAAGAT TGTTGAT 37

(2) INFORMATION FOR SEQ ID NO:139:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 35 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: Bovine respiratory syncytial virus

(B) STRAIN: Strain 375

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:139: CTCTGGATCC TACAGCCATG AGGATGATCA TCAGC 35

(2) INFORMATION FOR SEQ ID NO:140:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 40 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: Bovine respiratory syncytial virus

(B) STRAIN: Strain 375

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:140: CGTCGGATCC CTCACAGTTC CACATCATTG TCTTTGGGAT 40

(2) INFORMATION FOR SEQ ID NO:141:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 41 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: Bovine respiratory syncytial virus

(B) STRAIN: Strain 375

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:141: CTTAGGATCC CATGGCTCTT AGCAAGGTCA AACTAAATGA C 41

(2) INFORMATION FOR SEQ ID NO:142:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 41 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: Bovine respiratory syncytial virus

(B) STRAIN: Strain 375

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:142: CGTTGGATCC CTAGATCTGT GTAGTTGATT GATTTGTGTG A 41

(2) INFORMATION FOR SEQ ID NO:143:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 41 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: Bovine respiratory syncytial virus

(B) STRAIN: Strain 375

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:143: CTCTGGATCC TCATACCCAT CATCTTAAAT TCAAGACATT A 41

(2) INFORMATION FOR SEQ ID NO: 144:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 51 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:144: ACAGGAAACA GCTATGACCA TGATTACGAA TTCGAGCTCG CCCGGGGATC T 51

(2) INFORMATION FOR SEQ ID NO:145:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 128 base pairs (B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:145:

GTATAGCGGC CGCCTGCAGG TCGACTCTAG ATTTTTTTTT TTTTTTTTTT TGGCATATAA 60

ATAGATCTGT ATCCTAAAAT TGAATTGTAA TTATCGATAA TAAATGAATT TGATCCATGA 120

ATCCTAAT 128

(2) INFORMATION FOR SEQ ID NO:146:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 120 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:146: CTTTTCGTAA GGATCAATTC GGATCCATAA TTAATTAATT TTTATCCCGG CGCGCCTCGA 60 CTCTAGAATT TCATTTTGTT TTTTTCTATG CTATAAATGA ATTCGGATCC CGTCGTTTTA 120

(2) INFORMATION FOR SEQ ID NO:147:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 116 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:147: GAAATCCAGC TGAGCGCCGG TCGCTACCAT TACCAGTTGG TCTGGTGTCA AAAAGATCCA 60 TAATTAATTA ACCCGGGTCG AGGCGCGCCG GGTCGACCTG CAGGCGGCCG CTATAC 116

(2) INFORMATION FOR SEQ ID NO:148: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 51 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 148: TAATGTATCT ATAATGGTAT AAAGCTTGTA TTCTATAGTG TCACCTAAAT C 51

(2) INFORMATION FOR SEQ ID NO:149:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 51 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:149: ACAGGAAACA GCTATGACCA TGATTACGAA TTCGAGCTCG CCCGGGGATC T 51

(2) INFORMATION FOR SEQ ID NO: 150:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 168 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:150:

GTATTGCGGC CGCCTGCAGG TCGACTCTAG ATTTTTTTTT TTTTTTTTTT TGGCATATAA 60

ATAGATCTGT ATCCTAAAAT TGAATTGTAA TTATCGATAA TAAATGAATT CACCCGCTGG 120

TGGCGGTCTT TGGCGCGGGC CCCGTGGGCA TCGGCCCGGG CACCACGG 168

(2) INFORMATION FOR SEQ ID NO:151:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 112 base pairs (B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:151: GAGCTCGAAT TCGGATCCAT AATTAATTAA TTTTTATCCC GGCGCGCCTC GACTCTAGAA 60 TTTCATTTTG TTTTTTTCTA TGCTATAAAT GAATTCGGAT CCCGTCGTTT TA 112

(2) INFORMATION FOR SEQ ID NO:152:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 116 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:152: GAAATCCAGC TGAGCGCCGG TCGCTACCAT TACCAGTTGG TCTGGTGTCA AAAAGATCCA 60 TAATTAATTA ACCCGGGTCG AGGCGCGCCG GGTCGACCTG CAGGCGGCCG CTATAC 116

(2) INFORMATION FOR SEQ ID NO:153:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 51 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:153: TAATGTATCT ATAATGGTAT AAAGCTTGTA TTCTATAGTG TCACCTAAAT C 51

(2) INFORMATION FOR SEQ ID NO:154:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 51 base pairs

(B) TYPE: nucleic acid (C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:154: ACAGGAAACA GCTATGACCA TGATTACGAA TTCGAGCTCG CCCGGGGATC T 51

(2) INFORMATION FOR SEQ ID NO:155:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 104 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:155: AAATATATAA ATACCATGTT AGAATTTGGT CTGCTGCAGG TCGACTCTAG AATTTCATTT 60 TGTTTTTTTC TATGCTATAA ATGAATTCGG ATCCCGTCGT TTTA 104

(2) INFORMATION FOR SEQ ID NO:156:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 185 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:156:

GAAATCCAGC TGAGCGCCGG TCGCTACCAT TACCAGTTGG TCTGGTGTCA AAAAGATCCA 60

TAATTAATTA ACCCGGTCGA CTCTAGAAAA AATTGAAAAA CTATTCTAAT TTATTGCACG 120

GAGATCTTTT TTTTTTTTTT TTTTTTGGCA TATAAATGAA TTCGGATCCC CGGTGGCTTT 180

GGGGG 185

(2) INFORMATION FOR SEQ ID NO:157: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 66 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:157: CTCAATGTTA GGGTACCGAG CTCGAATTGG GTCGACCGGG TCGACCTGCA GCCTACATGG 60 AAATCT 66

(2) INFORMATION FOR SEQ ID NO:158:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 51 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:158: TAATGTATCT ATAATGGTAT AAAGCTTGTA TTCTATAGTG TCACCTAAAT C 51

(2) INFORMATION FOR SEQ ID NO:159:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 51 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:159: ACAGGAAACA GCTATGACCA TGATTACGAA TTCGAGCTCG CCCGGGGATC T 51

(2) INFORMATION FOR SEQ ID NO: 160:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 127 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:160:

GTATAGCGGC CGCCTGCAGG TCGACTCTAG ATTTTTTTTT TTTTTTTTTT TGGCATATAA 60

ATAGATCTGT ATCCTAAAAT TGAATTGTAA TTATCGATAA TAAATGAATT TCGACATGAA 120

TCCAAAT 127

(2) INFORMATION FOR SEQ ID NO:161:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 122 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:161:

GATAAGATGT AAGTCGAAAT TCGGATCCAT AATTAATTAA TTTTTATCCC GGCGCGCCTC 60

GACTCTAGAA TTTCATTTTG TTTTTTTCTA TGCTATAAAT GAATTCGGAT CCCGTCGTTT 120

TA 122

(2) INFORMATION FOR SEQ ID NO:162:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 116 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:162: GAAATCCAGC TGAGCGCCGG TCGCTACCAT TACCAGTTGG TCTGGTGTCA AAAAGATCCA 60 TAATTAATTA ACCCGGGTCG AGGCGCGCCG GGTCGACCTG CAGGCGGCCG CTATAC 116 (2) INFORMATION FOR SEQ ID NO:163:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 51 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:163: TAATGTATCT ATAATGGTAT AAAGCTTGTA TTCTATAGTG TCACCTAAAT C 51

(2) INFORMATION FOR SEQ ID NO:164:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 51 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:164: ACAGGAAACA GCTATGACCA TGATTACGAA TTCGAGCTCG CCCGGGGATC T 51

(2) INFORMATION FOR SEQ ID NO:165:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 61 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:165: GTATAGCGGC CGCCTGCAGG TCGACCTGCA GTGAATAATA AAATGTGTGT TTGTCCGAAA 60 T 61

(2) INFORMATION FOR SEQ ID NO:166:

(i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 45 base pairs (B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:166: CTCCATAGAA GACACCGGGA CCATGGATCC CGTCGTTTTA CAACG 45

(2) INFORMATION FOR SEQ ID NO:167:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 105 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:167: TCGGCGGAAA TCCAGCTGAG CGCCGGTCGC TACCATTACC AGTTGGTCTG GTGTCAAAAA 60 GATCTAGAAT AAGCTAGAGG ATCGATCCCC TATGGCGATC ATCAG 105

(2) INFORMATION FOR SEQ ID NO:168:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 31 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:168: CTGCAGGTCG ACCTGCAGGC GGCCGCTATA C 31

(2) INFORMATION FOR SEQ ID NO:169:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 51 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 169: TAATGTATCT ATAATGGTAT AAAGCTTGTA TTCTATAGTG TCACCTAAAT C 51

(2) INFORMATION FOR SEQ ID NO:170:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 51 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:170: ACAGGAAACA GCTATGACCA TGATTACGAA TTCGAGCTCG CCCGGGGATC T 51

(2) INFORMATION FOR SEQ ID NO:171:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 193 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:171:

GTATAGCGGC CGCCTGCAGG TCGACTCTAG ATTTTTTTTT TTTTTTTTTT TGGCATATAA 60

ATAGATCTGT ATCCTAAAAT TGAATTGTAA TTATCGATAA TAAATGAATT CCGAAGTGGG 120

CAACGTGGAT CCTCGCCCTC GGGCTCCTCG TGGTCCGCAC CGTCGTGGCC AGAAGTGCTC 180

CTACTAGCTC GAG 193

(2) INFORMATION FOR SEQ ID NO:172:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 123 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:172: ATCATTAGCA CGTTAACTTA ATAAGATCCA TAATTAATTA ATTTTTATCC CGGCGCGCCT 60 CGACTCTAGA ATTTCATTTT GTTTTTTTCT ATGCTATAAA TGAATTCGGA TCCCGTCGTT 120 TTA 123

(2) INFORMATION FOR SEQ ID NO:173:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 116 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:173: GAAATCCAGC TGAGCGCCGG TCGCTACCAT TACCAGTTGG TCTGGTGTCA AAAAGATCCA 60 TAATTAATTA ACCCGGGTCG AGGCGCGCCG GGTCGACCTG CAGGCGGCCG CTATAC 116

(2) INFORMATION FOR SEQ ID NO:174:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 51 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:174: TAATGTATCT ATAATGGTAT AAAGCTTGTA TTCTATAGTG TCACCTAAAT C 51

(2) INFORMATION FOR SEQ ID NO:175:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 51 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:175: ACAGGAAACA GCTATGACCA TGATTACGAA TTCGAGCTCG CCCGGGGATC T 51

(2) INFORMATION FOR SEQ ID NO:176:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 133 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO.-176:

GTATAGCGGC CGCCTGCAGG TCGACTCTAG ATTTTTTTTT _TTTTTTTTTT TGGCATATAA 60

ATAGATCTGT ATCCTAAAAT TGAATTGTAA TTATCGATAA TAAATGAATT CCTCTGGTTG 120

CCGTTCTGTC GGC 133

(2) INFORMATION FOR SEQ ID NO:177:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 99 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:177: GAAAATGAAA AAATGGTTTA AACCGGGGGC GCGCCTCGAC TCTAGAATTT CATTTTGTTT 60 TTTTCTATGC TATAAATGAA TTCGGATCCC GTCGTTTTA 99

(2) INFORMATION FOR SEQ ID NO:178:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 116 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:178: GAAATCCAGC TGAGCGCCGG TCGCTACCAT TACCAGTTGG TCTGGTGTCA AAAAGATCCA 60 TAATTAATTA ACCCGGGTCG AGGCGCGCCG GGTCGACCTG CAGGCGGCCG CTATAC 116

(2) INFORMATION FOR SEQ ID NO:179:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 51 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:179: TAATGTATCT ATAATGGTAT AAAGCTTGTA TTCTATAGTG TCACCTAAAT C 51

(2) INFORMATION FOR SEQ ID NO:180:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 51 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:180: ACAGGAAACA GCTATGACCA TGATTACGAA TTCGAGCTCG CCCGGGGATC T 51

(2) INFORMATION FOR SEQ ID NO:181:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 140 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 181: GTATAGCGGC CGCCTGCAGG TCGACTCTAG ATTTTTTTTT TTTTTTTTTT TGGCATATAA 60 ATAGATCTGT ATCCTAAAAT TGAATTGTAA TTATCGATAA TAAATGAATT CGGATCAGCT 120 TATGATGGAT GGACGTTTGG 140

(2) INFORMATION FOR SEQ ID NO: 182:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 123 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 182: GGAGGTGTCC ACGGCCTTAA AGCTGATCCA TAATTAATTA ATTTTTATCC CGGCGCGCCT 60 CGACTCTAGA ATTTCATTTT GTTTTTTTCT ATGCTATAAA TGAATTCGGA TCCCGTCGTT 120 TTA 123

(2) INFORMATION FOR SEQ ID NO:183:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 116 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:183: GAAATCCAGC TGAGCGCCGG TCGCTACCAT TACCAGTTGG TCTGGTGTCA AAAAGATCCA 60 TAATTAATTA ACCCGGGTCG AGGCGCGCCG GGTCGACCTG CAGGCGGCCG CTATAC 116

(2) INFORMATION FOR SEQ ID NO:184:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 51 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:184: TAATGTATCT ATAATGGTAT AAAGCTTGTA TTCTATAGTG TCACCTAAAT C 51

(2) INFORMATION FOR SEQ ID NO:185:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 39 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:185: GAAGCATGCC CGTTCTTATC AATAGTTTAG TCGAAAATA 39

(2) INFORMATION FOR SEQ ID NO:186:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 41 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:186: CATAAGATCT GGCATTGTGT TATTATACTA ACAAAAATAA G 41

(2) INFORMATION FOR SEQ ID NO:187:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 41 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (xi) SEQUENCE DESCRIPTION: SEQ ID NO:187: CCGTAGTCGA CAAAGATCGA CTTATTAATA TGTATGGGAT T 41

(2) INFORMATION FOR SEQ ID NO:188:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 39 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(Xi) SEQUENCE DESCRIPTION: SEQ ID NO:188: GCCTGAAGCT TCTAGTACAG TATTTACGAC TTTTGAAAT 39

(2) INFORMATION FOR SEQ ID NO:189:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 3942 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

(ix) FEATURE:

(A) NAME/KEY: CDS

(B) LOCATION: 1..369

(ix) FEATURE:

(A) NAME/KEY: CDS

(B) LOCATION: 370..597

(ix) FEATURE:

(A) NAME/KEY: CDS

(B) LOCATION: 598..1539

(ix) FEATURE:

(A) NAME/KEY: CDS

(B) LOCATION: 1675..3708

(ix) FEATURE:

(A) NAME/KEY: CDS

(B) LOCATION: complement (3748..3942)

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:189: TGT TTG TTC ATT AAT AAG ATG GGT GGA GCT ATT ATA GAA TAC AAG ATA 48

Cys Leu Phe He Asn Lys Met Gly Gly Ala He He Glu Tyr Lys He

1 5 10 15

CCT GGT TCC AAA TCT ATA ACC AAA TCT ATT TCC GAA GAA CTA GAA AAT 96 Pro Gly Ser Lys Ser He Thr Lys Ser He Ser Glu Glu Leu Glu Asn 20 25 30

TTA ACA AAG CGA GAT AAA CCA ATA TCT AAA ATT ATA GTT ATT CCT ATT 144 Leu Thr Lys Arg Asp Lys Pro He Ser Lys He He Val He Pro He 35 40 45

GTA TGT TAC AGA AAT GCA AAT AGT ATA AAG GTT ACA TTT GCA CTA AAA 192 Val Cys Tyr Arg Asn Ala Asn Ser He Lys Val Thr Phe Ala Leu Lys 50 55 60

AAG TTT ATC ATA GAT AAG GAG TTT AGT ACA AAT GTA ATA GAC GTA GAT 240 Lys Phe He He Asp Lys Glu Phe Ser Thr Asn Val He Asp Val Asp 65 70 75 80

GGT AAA CAT GAA AAA ATG TCC ATG AAT GAA ACA TGC GAA GAG GAT GTT 288 Gly Lys His Glu Lys Met Ser Met Asn Glu Thr Cys Glu Glu Asp Val 85 90 95

GCT AGA GGA TTG GGA ATT ATA GAT CTT GAA GAT GAA TGC ATA GAG GAA 336 Ala Arg Gly Leu Gly He He Asp Leu Glu Asp Glu Cys He Glu Glu 100 105 110

GAT GAT GTC GAT ACG TCA TTA TTT AAT GTA TAAATG GAT AAA TTG TAT 384 Asp Asp Val Asp Thr Ser Leu Phe Asn Val Met Asp Lys Leu Tyr 115 120 1 5

GCG GCA ATA TTC GGC GTT TTT ATG ACA TCT AAA GAT GAT GAT TTT AAT 432 Ala Ala He Phe Gly Val Phe Met Thr Ser Lys Asp Asp Asp Phe Asn 10 15 20

AAC TTT ATA GAA GTT GTA AAA TCT GTA TTA ACA GAT ACA TCA TCT AAT 480 Asn Phe He Glu Val Val Lys Ser Val Leu Thr Asp Thr Ser Ser Asn 25 30 35

CAT ACA ATA TCG TCG TCC AAT AAT AAT ACA TGG ATA TAT ATA TTT CTA 528 His Thr He Ser Ser Ser Asn Asn Asn Thr Trp He Tyr He Phe Leu 40 45 50

GCG ATA TTA TTT GGT GTT ATG GTA TTA TTA GTT TTT ATT TTG TAT TTA 576 Ala He Leu Phe Gly Val Met Val Leu Leu Val Phe He Leu Tyr Leu 55 60 65

AAA GTT ACT AAA CCA ACT TAAATG GAG GAA GCA GAT AAC CAA CTC GTT 624 Lys Val Thr Lys Pro Thr Met Glu Glu Ala Asp Asn Gin Leu Val 70 75 1 5

TTA AAT AGT ATT AGT GCT AGA GCA TTA AAG GCA TTT TTT GTA TCT AAA 672 Leu Asn Ser He Ser Ala Arg Ala Leu Lys Ala Phe Phe Val Ser Lys 10 15 20 25

ATT AAT GAT ATG GTC GAT GAA TTA GTT ACC AAA AAA TAT CCA CCA AAG 720 He Asn Asp Met Val Asp Glu Leu Val Thr Lys Lys Tyr Pro Pro Lys 30 35 40

AAG AAA TCA CAA ATA AAA CTC ATA GAT ACA CGA ATT CCT ATT GAT CTT 768 Lys Lys Ser Gin He Lys Leu He Asp Thr Arg He Pro He Asp Leu 45 50 55

ATT AAT CAA CAA TTC GTT AAA AGA TTT AAA CTA GAA AAT TAT AAA AAT 816 He Asn Gin Gin Phe Val Lys Arg Phe Lys Leu Glu Asn Tyr Lys Asn 60 65 70 GGA ATT TTA TCC GTT CTT ATC AAT AGT TTA GTC GAA AAT AAT TAC TTT 864 Gly He Leu Ser Val Leu He Asn Ser Leu Val Glu Asn Asn Tyr Phe 75 80 85

GAA CAA GAT GGT AAA CTT AAT AGC AGT GAT ATT GAT GAA TTA GTG CTC 912 Glu Gin Asp Gly Lys Leu Asn Ser Ser Asp He Asp Glu Leu Val Leu 90 95 100 105

ACA GAC ATA GAG AAA AAG ATT TTA TCG TTG ATT CCT AGA TGT TCT CCT 960 Thr Asp He Glu Lys Lys He Leu Ser Leu He Pro Arg Cys Ser Pro 110 115 120

CTT TAT ATA GAT ATC AGT GAC GTT AAA GTT CTC GCA TCT AGG TTA AAA 1008 Leu Tyr He Asp He Ser Asp Val Lys Val Leu Ala Ser Arg Leu Lys 125 130 135

AAA AGT GCT AAA TCA TTT ACG TTT AAT GAT CAT GAA TAT ATT ATA CAA 1056 Lys Ser Ala Lys Ser Phe Thr Phe Asn Asp His Glu Tyr He He Gin 140 145 150

TCT GAT AAA ATA GAG GAA TTA ATA AAT AGT TTA TCT AGA AAC CAT GAT 1104 Ser Asp Lys He Glu Glu Leu He Asn Ser Leu Ser Arg Asn His Asp 155 160 165

ATT ATA CTA GAT GAA AAA AGT TCT ATT AAA GAC AGC ATA TAT ATA CTA 1152 He He Leu Asp Glu Lys Ser Ser He Lys Asp Ser He Tyr He Leu 170 175 180 185

TCT GAT GAT CTT TTG AAT ATA CTT CGT GAA AGA TTA TTT AGA TGT CCA 1200 Ser Asp Asp Leu Leu Asn He Leu Arg Glu Arg Leu Phe Arg Cys Pro 190 195 200

CAG GTT AAA GAT AAT ACT ATT TCT AGA ACA CGT CTA TAT GAT TAT TTT 1248 Gin Val Lys Asp Asn Thr He Ser Arg Thr Arg Leu Tyr Asp Tyr Phe 205 210 215

ACT AGA GTG TCA AAG AAA GAA GAA GCG AAA ATA TAC GTT ATA TTG AAA 1296 Thr Arg Val Ser Lys Lys Glu Glu Ala Lys He Tyr Val He Leu Lys 220 225 230

GAT TTA AAG ATT GCT GAT ATA CTC GGT ATC GAA ACA GTA ACG ATA GGA 1344 Asp Leu Lys He Ala Asp He Leu Gly He Glu Thr Val Thr He Gly 235 240 245

TCA TTT GTA TAT ACG AAA TAT AGC ATG TTG ATT AAT TCA ATT TCG TCT 1392 Ser Phe Val Tyr Thr Lys Tyr Ser Met Leu He Asn Ser He Ser Ser 250 255 260 265

AAT GTT GAT AGA TAT TCA AAA AGG TTC CAT GAC TCT TTT TAT GAA GAT 1440 Asn Val Asp Arg Tyr Ser Lys Arg Phe His Asp Ser Phe Tyr Glu Asp 270 275 280

ATT GCG GAA TTT ATA AAG GAT AAT GAA AAA ATT AAT GTA TCC AGA GTT 1488 He Ala Glu Phe He Lys Asp Asn Glu Lys He Asn Val Ser Arg Val 285 290 295

GTT GAA TGC CTT ATC GTA CCT AAT ATT AAT ATA GAG TTA TTA ACT GAA 1536 Val Glu Cys Leu He Val Pro Asn He Asn He Glu Leu Leu Thr Glu 300 305 310

TAAGTATATA TAAATGATTG TTTTTATAAT GTTTGTTATC GCATTTAGTT TTGCTGTATG 1596

GTTATCATAT ACATTTTTAA GGCCGTATAT GATAAATGAA AATATATAAG CACTTATTTT 1656

TGTTAGTATA ATAACACA ATG CCG TCG TAT ATG TAT CCG AAG AAC GCA AGA 1707

Met Pro Ser Tyr Met Tyr Pro Lys Asn Ala Arg 1 5 10 AAA GTA ATT TCA AAG ATT ATA TCA TTA CAA CTT GAT ATT AAA AAA CTT 1755 Lys Val He Ser Lys He He Ser Leu Gin Leu Asp He Lys Lys Leu 15 20 25

CCT AAA AAA TAT ATA AAT ACC ATG TTA GAA TTT GGT CTA CAT GGA AAT 1803 Pro Lys Lys Tyr He Asn Thr Met Leu Glu Phe Gly Leu His Gly Asn 30 35 40

CTA CCA GCT TGT ATG TAT AAA GAT GCC GTA TCA TAT GAT ATA AAT AAT 1851 Leu Pro Ala Cys Met Tyr Lys Asp Ala Val Ser Tyr Asp He Asn Asn 45 50 55

ATA AGA TTT TTA CCT TAT AAT TGT GTT ATG GTT AAA GAT TTA ATA AAT 1899 He Arg Phe Leu Pro Tyr Asn Cys Val Met Val Lys Asp Leu He Asn 60 65 70 75

GTT ATA AAA TCA TCA TCT GTA ATA GAT ACT AGA TTA CAT CAA TCT GTA 1947 Val He Lys Ser Ser Ser Val He Asp Thr Arg Leu His Gin s^'er Val 80 85 90

TTA AAA CAT CGT AGA GCG TTA ATA GAT TAC GGC GAT CAA GAC ATT ATC 1995 Leu Lys His Arg Arg Ala Leu He Asp Tyr Gly Asp Gin Asp He He 95 100 105

ACT TTA ATG ATC ATT AAT AAG TTA CTA TCG ATA GAT GAT ATA TCC TAT 2043 Thr Leu Met He He Asn Lys Leu Leu Ser He Asp Asp He Ser Tyr 110 115 120

ATA TTA GAT AAA AAA ATA ATT CAT GTA ACA AAA ATA TTA AAA ATA GAC 2091 He Leu Asp Lys Lys He He His Val Thr Lys He Leu Lys He Asp 125 130 135

CCT ACA GTA GCC AAT TCA AAC ATG AAA CTG AAT AAG ATA GAG CTT GTA 2139 Pro Thr Val Ala Asn Ser Asn Met Lys Leu Asn Lys He Glu Leu Val 140 145 150 155

GAT GTA ATA ACA TCA ATA CCT AAG TCT TCC TAT ACA TAT TTA TAT AAT 2187 Asp Val He Thr Ser He Pro Lys Ser Ser Tyr Thr Tyr Leu Tyr Asn 160 165 170

AAT ATG ATC ATT GAT CTC GAT ACA TTA TTA TAT TTA TCC GAT GCA TTC 2235 Asn Met He He Asp Leu Asp Thr Leu Leu Tyr Leu Ser Asp Ala Phe 175 180 185

CAC ATA CCC CCC ACA CAT ATA TCA TTA CGT TCA CTT AGA GAT ATA AAC 2283 His He Pro Pro Thr His He Ser Leu Arg Ser Leu Arg Asp He Asn 190 195 200

AGG ATT ATT GAA TTG CTT AAA AAA TAT CCG AAT AAT AAT ATT ATT GAT 2331 Arg He He Glu Leu Leu Lys Lys Tyr Pro Asn Asn Asn He He Asp 205 210 215

TAT ATA TCC GAT AGC ATA AAA TCA AAT AGT TCA TTC ATT CAC ATA CTT 2379 Tyr He Ser Asp Ser He Lys Ser Asn Ser Ser Phe He His He Leu 220 225 230 235

CAT ATG ATA ATA TCA AAT ATG TTT CCT GCT ATA ATC CCT AGT GTA AAC 2427 His Met He He Ser Asn Met Phe Pro Ala He He Pro Ser Val Asn 240 245 250

GAT TTT ATA TCT ACC GTA GTT GAT AAA GAT CGA CTT ATT AAT ATG TAT 2475 Asp Phe He Ser Thr Val Val Asp Lys Asp Arg Leu He Asn Met Tyr 255 260 265

GGG ATT AAG TGT GTT GCT ATG TTT TCG TAC GAT ATA AAC ATG ATC GAT 2523 Gly He Lys Cys Val Ala Met Phe Ser Tyr Asp He Asn Met He Asp 270 275 280 TTA GAG TCA TTA GAT GAC TCA GAT TAC ATA TTT ATA GAA AAA AAT ATA 2571 Leu Glu Ser Leu Asp Asp Ser Asp Tyr He Phe He Glu Lys Asn He 285 290 295

TCT ATA TAC GAC GTT AAA TGT AGA GAT TTT GCG AAT ATG ATT AGA GAT 2619 Ser He Tyr Asp Val Lys Cys Arg Asp Phe Ala Asn Met He Arg Asp 300 305 310 315

AAG GTT AAA AGA GAA AAG AAT AGA ATA TTA ACT ACG AAA TGT GAA GAT 2667 Lys Val Lys Arg Glu Lys Asn Arg He Leu Thr Thr Lys Cys Glu Asp 320 325 330

ATT ATA AGA TAT ATA AAA TTA TTC AGT AAA AAT AGA ATA AAC GAT GAA 2715 He He Arg Tyr He Lys Leu Phe Ser Lys Asn Arg He Asn Asp Glu 335 340 345

AAT AAT AAG GTG GAG GAG GTG TTG ATA CAT ATT GAT AAT GTA TCT AAA 2763 Asn Asn Lys Val Glu Glu Val Leu He His He Asp Asn Val Ser Lys 350 355 360

AAT AAT AAA TTA TCA CTG TCT GAT ATA TCA TCT TTA ATG GAT CAA TTT 2811 Asn Asn Lys Leu Ser Leu Ser Asp He Ser Ser Leu Met Asp Gin Phe 365 370 375

CGT TTA AAT CCA TGT ACC ATA AGA AAT ATA TTA TTA TCT TCA GCA ACT 2859 Arg Leu Asn Pro Cys Thr He Arg Asn He Leu Leu Ser Ser Ala Thr 380 385 390 395

ATA AAA TCA AAA CTA TTA GCG TTA CGG GCA GTA AAA AAC TGG AAA TGT 2907 He Lys Ser Lys Leu Leu Ala Leu Arg Ala Val Lys Asn Trp Lys Cys 400 405 410

TAT TCA TTG ACA AAT GTA TCA ATG TAT AAA AAA ATA AAG GGT GTT ATC 2955 Tyr Ser Leu Thr Asn Val Ser Met Tyr Lys Lys He Lys Gly Val He 415 420 425

GTA ATG GAT ATG GTT GAT TAT ATA TCT ACT AAC ATT CTT AAA TAC CAT 3003 Val Met Asp Met Val Asp Tyr He Ser Thr Asn He Leu Lys Tyr His 430 435 440

AAA CAA TTA TAT GAT AAA ATG AGT ACG TTT GAA TAT AAA CGA GAT ATT 3051 Lys Gin Leu Tyr Asp Lys Met Ser Thr Phe Glu Tyr Lys Arg Asp He 445 450 455

AAA TCA TGT AAA TGC TCG ATA TGT TCC GAC TCT ATA ACA CAT CAT ATA 3099 Lys Ser Cys Lys Cys Ser He Cys Ser Asp Ser He Thr His His He 460 465 470 475

TAT GAA ACA ACA TCA TGT ATA AAT TAT AAA TCT ACC GAT AAT GAT CTT 3147 Tyr Glu Thr Thr Ser Cys He Asn Tyr Lys Ser Thr Asp Asn Asp Leu 480 485 490

ATG ATA GTA TTG TTC AAT CTA ACT AGA TAT TTA ATG CAT GGG ATG ATA 3195 Met He Val Leu Phe Asn Leu Thr Arg Tyr Leu Met His Gly Met He 495 500 505

CAT CCT AAT CTT ATA AGC GTA AAA GGA TGG GGT CCC CTT ATT GGA TTA 3243 His Pro Asn Leu He Ser Val Lys Gly Trp Gly Pro Leu He Gly Leu 510 515 520

TTA ACG GGT GAT ATA GGT ATT AAT TTA AAA CTA TAT TCC ACC ATG AAT 3291 Leu Thr Gly Asp He Gly He Asn Leu Lys Leu Tyr Ser Thr Met Asn 525 530 535

ATA AAT GGG CTA CGG TAT GGA GAT ATT ACG TTA TCT TCA TAC GAT ATG 3339 He Asn Gly Leu Arg Tyr Gly Asp He Thr Leu Ser Ser Tyr Asp Met 540 545 550 555 AGT AAT AAA TTA GTC TCT ATT ATT AAT ACA CCC ATA TAT GAG TTA ATA 3387 Ser Asn Lys Leu Val Ser He He Asn Thr Pro He Tyr Glu Leu He 560 565 570

CCG TTT ACT ACA TGT TGT TCA CTC AAT GAA TAT TAT TCA AAA ATT GTG 3435 Pro Phe Thr Thr Cys Cys Ser Leu Asn Glu Tyr Tyr Ser Lys He Val 575 580 585

ATT TTA ATA AAT GTT ATT TTA GAA TAT ATG ATA TCT ATT ATA TTA TAT 3483 He Leu He Asn Val He Leu Glu Tyr Met He Ser He He Leu Tyr 590 595 600

AGA ATA TTG ATC GTA AAA AGA TTT AAT AAC ATT AAA GAA TTT ATT TCA 3531 Arg He Leu He Val Lys Arg Phe Asn Asn He Lys Glu Phe He Ser 605 610 615

AAA GTC GTA AAT ACT GTA CTA GAA TCA TCA GGC ATA TAT TTT TGT CAG 3579 Lys Val Val Asn Thr Val Leu Glu Ser Ser Gly He Tyr Phe Cys Gin 620 625 630 635

ATG CGT GTA CAT GAA CAA ATT GAA TTG GAA ATA GAT GAG CTC ATT ATT 3627 Met Arg Val His Glu Gin He Glu Leu Glu He Asp Glu Leu He He 640 645 650

AAT GGA TCT ATG CCT GTA CAG CTT ATG CAT TTA CTT CTA AAG GTA GCT 3675 Asn Gly Ser Met Pro Val Gin Leu Met His Leu Leu Leu Lys Val Ala 655 660 665

ACC ATA ATA TTA GAG GAA ATC AAA GAA ATA TAACGTATTT TTTCTTTTAA 3725 Thr He He Leu Glu Glu He Lys Glu He 670 675

ATAAATAAAA ATACTTTTTT TTTTAAACAA GGGGTGCTAC CTTGTCTAAT TGTATCTTGT 3785

ATTTTGGATC TGATGCAAGA TTATTAAATA ATCGTATGAA AAAGTAGTAG ATATAGTTTA 3845

TATCGTTACT GGACATGATA TTATGTTTAG TTAATTCTTC TTTGGCATGA ATTCTACACG 3905

TCGGACAAGG TAATGTATCT ATAATGGTAT AAAGCTT 3942

(2) INFORMATION FOR SEQ ID NO:190:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 122 amino acids

(B) TYPE: amino acid (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: peptide

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:190:

Cys Leu Phe He Asn Lys Met Gly Gly Ala He He Glu Tyr Lys He 1 5 10 15

Pro Gly Ser Lys Ser He Thr Lys Ser He Ser Glu Glu Leu Glu Asn 20 25 30

Leu Thr Lys Arg Asp Lys Pro He Ser Lys He He Val He Pro He 35 40 45

Val Cys Tyr Arg Asn Ala Asn Ser He Lys Val Thr Phe Ala Leu Lys 50 55 60

Lys Phe He He Asp Lys Glu Phe Ser Thr Asn Val He Asp Val Asp 65 70 75 80 Gly Lys His Glu Lys Met Ser Met Asn Glu Thr Cys Glu Glu Asp Val 85 90 95

Ala Arg Gly Leu Gly He He Asp Leu Glu Asp Glu Cys He Glu Glu 100 105 110

Asp Asp Val Asp Thr Ser Leu Phe Asn Val 115 120

(2) INFORMATION FOR SEQ ID NO:191:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 75 amino acids

(B) TYPE: amino acid (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:191:

Met Asp Lys Leu Tyr Ala Ala He Phe Gly Val Phe Met Thr Ser Lys 1 5 10 15

Asp Asp Asp Phe Asn Asn Phe He Glu Val Val Lys Ser Val Leu Thr 20 25 30

Asp Thr Ser Ser Asn His Thr He Ser Ser Ser Asn Asn Asn Thr Trp 35 40 45

He Tyr He Phe Leu Ala He Leu Phe Gly Val Met Val Leu Leu Val 50 55 60

Phe He Leu Tyr Leu Lys Val Thr Lys Pro Thr 65 70 75

(2) INFORMATION FOR SEQ ID NO:192:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 313 amino acids

(B) TYPE: amino acid (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

( i) SEQUENCE DESCRIPTION: SEQ ID NO:192:

Met Glu Glu Ala Asp Asn Gin Leu Val Leu Asn Ser He Ser Ala Arg 1 5 10 15

Ala Leu Lys Ala Phe Phe Val Ser Lys He Asn Asp Met Val Asp Glu 20 25 30

Leu Val Thr Lys Lys Tyr Pro Pro Lys Lys Lys Ser Gin He Lys Leu 35 40 45

He Asp Thr Arg He Pro He Asp Leu He Asn Gin Gin Phe Val Lys 50 55 60

Arg Phe Lys Leu Glu Asn Tyr Lys Asn Gly He Leu Ser Val Leu He 65 70 75 80

Asn Ser Leu Val Glu Asn Asn Tyr Phe Glu Gin Asp Gly Lys Leu Asn 85 90 95 Ser Ser Asp He Asp Glu Leu Val Leu Thr Asp He Glu Lys Lys He 100 105 110

Leu Ser Leu He Pro Arg Cys Ser Pro Leu Tyr He Asp He Ser Asp 115 120 125

Val Lys Val Leu Ala Ser Arg Leu Lys Lys Ser Ala Lys Ser Phe Thr 130 135 140

Phe Asn Asp His Glu Tyr He He Gin Ser Asp Lys He Glu Glu Leu 145 150 155 160

He Asn Ser Leu Ser Arg Asn His Asp He He Leu Asp Glu Lys Ser 165 170 175

Ser He Lys Asp Ser He Tyr He Leu Ser Asp Asp Leu Leu Asn He 180 185 190

Leu Arg Glu Arg Leu Phe Arg Cys Pro Gin Val Lys Asp Asn Thr He 195 200 205

Ser Arg Thr Arg Leu Tyr Asp Tyr Phe Thr Arg Val Ser Lys Lys Glu 210 215 220

Glu Ala Lys He Tyr Val He Leu Lys Asp Leu Lys He Ala Asp He 225 230 235 240

Leu Gly He Glu Thr Val Thr He Gly Ser Phe Val Tyr Thr Lys Tyr 245 250 255

Ser Met Leu He Asn Ser He Ser Ser Asn Val Asp Arg Tyr Ser Lys 260 265 270

Arg Phe His Asp Ser Phe Tyr Glu Asp He Ala Glu Phe He Lys Asp 275 280 285

Asn Glu Lys He Asn Val Ser Arg Val Val Glu Cys Leu He Val Pro 290 295 300

Asn He Asn He Glu Leu Leu Thr Glu 305 310

(2) INFORMATION FOR SEQ ID NO:193:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 677 amino acids

(B) TYPE: amino acid (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:193:

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 Thr Lys He Leu Lys He Asp Pro Thr Val Ala Asn 130 135 140

Ser Asn Met Lys Leu Asn Lys He Glu Leu Val Asp Val He Thr Ser 145 150 155 160

He Pro Lys Ser Ser Tyr Thr Tyr Leu Tyr Asn Asn Met He He Asp 165 170 175

Leu Asp Thr Leu Leu Tyr Leu Ser Asp Ala Phe His He Pro Pro Thr 180 185 190

His He Ser Leu Arg Ser Leu Arg Asp He Asn Arg He He Glu Leu 195 200 205

Leu Lys Lys Tyr Pro Asn Asn Asn He He Asp Tyr He Ser Asp Ser 210 215 220

He Lys Ser Asn Ser Ser Phe He His He Leu His Met He He Ser 225 230 235 240

Asn Met Phe Pro Ala He He Pro Ser Val Asn Asp Phe He Ser Thr 245 250 255

Val Val Asp Lys Asp Arg Leu He Asn Met Tyr Gly He Lys Cys Val 260 265 270

Ala Met Phe Ser Tyr Asp He Asn Met He Asp Leu Glu Ser Leu Asp 275 280 285

Asp Ser Asp Tyr He Phe He Glu Lys Asn He Ser He Tyr Asp Val 290 295 300

Lys Cys Arg Asp Phe Ala Asn Met He Arg Asp Lys Val Lys Arg Glu 305 310 315 320

Lys Asn Arg He Leu Thr Thr Lys Cys Glu Asp He He Arg Tyr He 325 330 335

Lys Leu Phe Ser Lys Asn Arg He Asn Asp Glu Asn Asn Lys Val Glu 340 345 350

Glu Val Leu He His He Asp Asn Val Ser Lys Asn Asn Lys Leu Ser 355 360 365

Leu Ser Asp He Ser Ser Leu Met Asp Gin Phe Arg Leu Asn Pro Cys 370 375 380

Thr He Arg Asn He Leu Leu Ser Ser Ala Thr He Lys Ser Lys Leu 385 390 395 400

Leu Ala Leu Arg Ala Val Lys Asn Trp Lys Cys Tyr Ser Leu Thr Asn 405 410 415 Val Ser Met Tyr Lys Lys He Lys Gly Val He Val Met Asp Met Val 420 425 430

Asp Tyr He Ser Thr Asn He Leu Lys Tyr His Lys Gin Leu Tyr Asp 435 440 445

Lys Met Ser Thr Phe Glu Tyr Lys Arg Asp He Lys Ser Cys Lys Cys 450 455 460

Ser He Cys Ser Asp Ser He Thr His His He Tyr Glu Thr Thr Ser 465 470 475 480

Cys He Asn Tyr Lys Ser Thr Asp Asn Asp Leu Met He Val Leu Phe 485 490 495

Asn Leu Thr Arg Tyr Leu Met His Gly Met He His Pro Asn Leu He 500 505 510

Ser Val Lys Gly Trp Gly Pro Leu He Gly Leu Leu Thr Gly Asp He 515 520 525

Gly He Asn Leu Lys Leu Tyr Ser Thr Met Asn He Asn Gly Leu Arg 530 535 540

Tyr Gly Asp He Thr Leu Ser Ser Tyr Asp Met Ser Asn Lys Leu Val 545 550 555 560

Ser He He Asn Thr Pro He Tyr Glu Leu He Pro Phe Thr Thr Cys 565 570 575

Cys Ser Leu Asn Glu Tyr Tyr Ser Lys He Val He Leu He Asn Val 580 585 590

He Leu Glu Tyr Met He Ser He He Leu Tyr Arg He Leu He Val 595 600 605

Lys Arg Phe Asn Asn He Lys Glu Phe He Ser Lys Val Val Asn Thr 610 615 620

Val Leu Glu Ser Ser Gly He Tyr Phe Cys Gin Met Arg Val His Glu 625 630 635 640

Gin He Glu Leu Glu He Asp Glu Leu He He Asn Gly Ser Met Pro 645 650 655

Val Gin Leu Met His Leu Leu Leu Lys Val Ala Thr He He Leu Glu 660 665 670

Glu He Lys Glu He 675

(2) INFORMATION FOR SEQ ID NO:194:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 64 amino acids

(B) TYPE: amino acid (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

( i) SEQUENCE DESCRIPTION: SEQ ID NO:194:

Lys Leu Tyr Thr He He Asp Thr Leu Pro Cys Pro Thr Cys Arg He 1 5 10 15 His Ala Lys Glu Glu Leu Thr Lys His Asn He Met Ser Ser Asn Asp 20 25 30

He Asn Tyr He Tyr Tyr Phe Phe He Arg Leu Phe Asn Asn Leu Ala 35 40 45

Ser Asp Pro Lys Tyr Lys He Gin Leu Asp Lys Val Ala Pro Leu Val 50 55 60

(2) INFORMATION FOR SEQ ID NO:195:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 583 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

(ix) FEATURE:

(A) NAME/KEY: CDS

(B) LOCATION: 2..583

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:195:

A AGC TTA AGA AAG AAT GTA GGG AAC GAA GAA TAT AGA ACC AAA GAT 46 Ser Leu Arg Lys Asn Val Gly Asn Glu Glu Tyr Arg Thr Lys Asp 1 5 10 15

TTA TTT ACT GCA TTA TGG GTA CCT GAT TTA TTT ATG GAA CGC GTA GAA 94 Leu Phe Thr Ala Leu Trp Val Pro Asp Leu Phe Met Glu Arg Val Glu 20 25 30

AAA GAT GAA GAA TGG TCT CTA ATG TGT CCA TGC GAA TGT CCA GGA TTA 142 Lys Asp Glu Glu Trp Ser Leu Met Cys Pro Cys Glu Cys Pro Gly Leu 35 40 45

TGC GAT GTA TGG GGG AAT GAT TTT AAC AAA TTA TAT ATA GAA TAC GAA 190 Cys Asp Val Trp Gly Asn Asp Phe Asn Lys Leu Tyr He Glu Tyr Glu 50 55 60

ACA AAG AAA AAA ATT AAA GCG ATC GCT AAA GCA AGA AGT TTA TGG AAA 238 Thr Lys Lys Lys He Lys Ala He Ala Lys Ala Arg Ser Leu Trp Lys 65 70 75

TCT ATT ATC GAG GCT CAA ATA GAA CAA GGA ACG CCG TAT ATA CTA TAT 286 Ser He He Glu Ala Gin He Glu Gin Gly Thr Pro Tyr He Leu Tyr 80 85 90 95

AAA GAT TCT TGT AAT AAA AAA TCC AAT CAA AGC AAT TTG GGA ACA ATT 334 Lys Asp Ser Cys Asn Lys Lys Ser Asn Gin Ser Asn Leu Gly Thr He 100 105 110

AGA TCG AGT AAT CTC TGT ACA GAG ATT ATA CAA TTT AGT AAC GAG GAT 382 Arg Ser Ser Asn Leu Cys Thr Glu He He Gin Phe Ser Asn Glu Asp 115 120 125 GAA GTT GCT GTA TGT AAT CTA GGA TCT ATT TCG TGG AGT AAA TTT GTT 430 Glu Val Ala Val Cys Asn Leu Gly Ser He Ser Trp Ser Lys Phe Val 130 135 140

AAT AAT AAC GTA TTT ATG TTC GAC AAG TTG AGA ATA ATT ACG AAA ATA 478 Asn Asn Asn Val Phe Met Phe Asp Lys Leu Arg He He Thr Lys He 145 150 155

CTA GTT AAA AAT CTA AAT AAA ATA ATA GAT ATC AAT TAT TAT CCA GTG 526 Leu Val Lys Asn Leu Asn Lys He He Asp He Asn Tyr Tyr Pro Val 160 165 170 175

ATA GAA TCG TCT AGA TCT AAT AAG AAA CAT AGA CCC ATA GGT ATC GGG 574 He Glu Ser Ser Arg Ser Asn Lys Lys His Arg Pro He Gly He Gly 180 185 190

GTT CAG GGT 583

Val Gin Gly

(2) INFORMATION FOR SEQ ID NO:196:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 194 amino acids

(B) TYPE: amino acid (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:196:

Ser Leu Arg Lys Asn Val Gly Asn Glu Glu Tyr Arg Thr Lys Asp Leu 1 5 10 15

Phe Thr Ala Leu Trp Val Pro Asp Leu Phe Met Glu Arg Val Glu Lys 20 25 30

Asp Glu Glu Trp Ser Leu Met Cys Pro Cys Glu Cys Pro Gly Leu Cys 35 40 45

Asp Val Trp Gly Asn Asp Phe Asn Lys Leu Tyr He Glu Tyr Glu Thr 50 55 60

Lys Lys Lys He Lys Ala He Ala Lys Ala Arg Ser Leu Trp Lys Ser 65 70 75 80

He He Glu Ala Gin He Glu Gin Gly Thr Pro Tyr He Leu Tyr Lys 85 90 95

Asp Ser Cys Asn Lys Lys Ser Asn Gin Ser Asn Leu Gly Thr He Arg 100 105 110

Ser Ser Asn Leu Cys Thr Glu He He Gin Phe Ser Asn Glu Asp Glu 115 120 125

Val Ala Val Cys Asn Leu Gly Ser He Ser Trp Ser Lys Phe Val Asn 130 135 140

Asn Asn Val Phe Met Phe Asp Lys Leu Arg He He Thr Lys He Leu 145 150 155 160

Val Lys Asn Leu Asn Lys He He Asp He Asn Tyr Tyr Pro Val He 165 170 175

Glu Ser Ser Arg Ser Asn Lys Lys His Arg Pro He Gly He Gly Val 180 185 190 Gln Gly

(2) INFORMATION FOR SEQ ID NO:197:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 51 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 197: ACAGGAAACA GCTATGACCA TGATTACGAA TTCGAGCTCG CCCGGGGATC T 51

(2) INFORMATION FOR SEQ ID NO:198:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 138 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:198:

GTATAGCGGC CGCCTGCAGG TCGACTCTAG ATTTTTTTTT TTTTTTTTTT TGGCATATAA 60

ATAGATCTGT ATCCTAAAAT TGAATTGTAA TTATCGATAA TAAATGAATT CGATGGCTGT 120

GCCTGCAAGC CCACAGCA 138

(2) INFORMATION FOR SEQ ID NO: 199:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 120 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic)

(iii) HYPOTHETICAL: NO

(iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:199: CTTAGCCCCA AACGCACCTC AGATCCATAA TTAATTAATT TTTATCCCGG CGCGCCTCGA 60 CTCTAGAATT TCATTTTGTT TTTTTCTATG CTATAAATGA ATTCGGATCC CGTCGTTTTA 120

(2) INFORMATION FOR SEQ ID NO:200:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 116 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:200: GAAATCCAGC TGAGCGCCGG TCGCTACCAT TACCAGTTGG TCTGGTGTCA AAAAGATCCA 60 TAATTAATTA ACCCGGGTCG AGGCGCGCCG GGTCGACCTG CAGGCGGCCG CTATAC 116

(2) INFORMATION FOR SEQ ID NO:201:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 51 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:201: TAATGTATCT ATAATGGTAT AAAGCTTGTA TTCTATAGTG TCACCTAAAT C 51

(2) INFORMATION FOR SEQ ID NO:202:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 51 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:202: ACAGGAAACA GCTATGACCA TGATTACGAA TTCGAGCTCG CCCGGGGATC T 51 (2) INFORMATION FOR SEQ ID NO:203:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 141 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:203:

GTATAGCGGC CGCCTGCAGG TCGACTCTAG ATTTTTTTTT TTTTTTTTTT TGGCATATAA 60

ATAGATCTGT ATCCTAAAAT TGAATTGTAA TTATCGATAA TAAATGAATT CCATGTGCTG 120

CCTCACCCCT GTGCTGGCGC T 141

(2) INFORMATION FOR SEQ ID NO:204:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 120 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:204: TCGCCCGCCT CTGACGCCCC GGATCCATAA TTAATTAATT TTTATCCCGG CGCGCCTCGA 60 CTCTAGAATT TCATTTTGTT TTTTTCTATG CTATAAATGA ATTCGGATCC CGTCGTTTTA 120

(2) INFORMATION FOR SEQ ID NO:205:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 116 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:205: GAAATCCAGC TGAGCGCCGG TCGCTACCAT TACCAGTTGG TCTGGTGTCA AAAAGATCCA 60 TAATTAATTA ACCCGGGTCG AGGCGCGCCG GGTCGACCTG CAGGCGGCCG CTATAC 116 (2) INFORMATION FOR SEQ ID NO:206:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 51 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:206: TAATGTATCT ATAATGGTAT AAAGCTTGTA TTCTATAGTG TCACCTAAAT C 51

(2) INFORMATION FOR SEQ ID NO:207:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 45 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:207: CAAGGAATGG TGCATGCCCG TTCTTATCAA TAGTTTAGTC GAAAA 45

(2) INFORMATION FOR SEQ ID NO:208:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 57 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:208: TATATAAGCA CTTATTTTTG TTAGTATAAT AACACAATGC CAGATCCCGT CGTTTTA 57

(2) INFORMATION FOR SEQ ID NO:209: (i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 249 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(Xi) SEQUENCE DESCRIPTION: SEQ ID NO:209:

TCCAGCTGAG CGCCGGTCGC TACCATTACC AGTTGGTCTG GTGTCAAAAA GATCCATAAT 60

TAATTAACCA GCGGCCGCCT GCAGGTCGAC TCTAGATTTT TTTTTTTTTT TTTTTTGGCA 120

TATAAATAGA TCTGTATCCT AAAATTGAAT TGTAATTATC GATAATAAAT GAATTCGGAT 180

CCATAATTAA TTAATTTTTA TCCCGGCGCG CCGGGTCGAC CTGCAGGCGG CCGCTGGGTC 240

GACAAAGAT 249

(2) INFORMATION FOR SEQ ID NO:210:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 45 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:210: CAAAAGTCGT AAATACTGTA CTAGAAGCTT GGCGTAATCA TGGTC 45

(2) INFORMATION FOR SEQ ID NO:211:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 33 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:211: CGACGGATCC GAGGTGCGTT TGGGGCTAAG TGC 33

(2) INFORMATION FOR SEQ ID NO:212: (i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 36 base pairs

(B) TYPE: nucleic acid (C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:212: CCACGGATCC AGCACAACGC GAGTCCCACC ATGGCT 36

(2) INFORMATION FOR SEQ ID NO:213:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 35 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:213: CCACGAATTC GATGGCTGTG CCTGCAAGCC CACAG 35

(2) INFORMATION FOR SEQ ID NO:214:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 32 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:214: CGAAGATCTG AGGTGCGTTT GGGGCTAAGT GC 32

(2) INFORMATION FOR SEQ ID NO:215:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 34 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:215: CGCAGGATCC GGGGCGTCAG AGGCGGGCGA GGTG 34

(2) INFORMATION FOR SEQ ID NO:216:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 32 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:216: GAGCGGATCC TGCAGGAGGA GACACAGAGC TG 32

(2) INFORMATION FOR SEQ ID NO:217:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 32 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(Xi) SEQUENCE DESCRIPTION: SEQ ID NO:217: GCGCGAATTC CATGTGCTGC CTCACCCCTG TG 32

(2) INFORMATION FOR SEQ ID NO:218:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 34 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO (xi) SEQUENCE DESCRIPTION: SEQ ID NO:218: CGCAGGATCC GGGGCGTCAG AGGCGGGCGA GGTG 34

(2) INFORMATION FOR SEQ ID NO:219:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 32 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:219: GGGGAATTCA ATGCAACCCA CCGCGCCGCC CC 32

(2) INFORMATION FOR SEQ ID NO:2 0:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 32 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: double

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iv) ANTI-SENSE: NO

(Xi) SEQUENCE DESCRIPTION: SEQ ID NO:220: GGGGATCCT AGGGCGCGCC CGCCGGCTCG CT 32

(2) INFORMATION FOR SEQ ID NO:221:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 5785 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: N (iv) ANTI-SENSE: N

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:221: AAGCTTAAGA AAGAATGTAG GGAACGAAGA ATATAGAACC AAAGATTTAT TTACTGCATT 60 ATGGGTACCT GATTTATTTA TGGAACGCGT AGAAAAAGAT GAAGAATGGT CTCTAATGTG 120 TCCATGCGAA TGTCCAGGAT TATGCGATGT ATGGGGGAAT GATTTTAACA AATTATATAT 180

AGAATACGAA ACAAAGAAAA AAATTAAAGC GATCGCTAAA GCAAGAAGTT TATGGAAATC 240

TATTATCGAG GCTCAAATAG AACAAGGAAC GCCGTATATA CTATATAAAG ATTCTTGTAA 300

TAAAAAATCC AATCAAAGCA ATTTGGGAAC AATTAGATCG AGTAATCTCT GTACAGAGAT 360

TATACAATTT AGTAACGAGG ATGAAGTTGC TGTATGTAAT CTAGGATCTA TTTCGTGGAG 420

TAAATTTGTT AATAATAACG TATTTATGTT CGACAAGTTG AGAATAATTA CGAAAATACT 480

AGTTAAAAAT CTAAATAAAA TAATAGATAT CAATTATTAT CCAGTGATAG AATCGTCTAG 540

ATCTAATAAG AAACATAGAC CCATAGGTAT CGGTGTTCAG GGTTTGGCTG ATGTGTTTAT 600

ATTATTGGGC TATGCATTCG ATAGCGAAGA AGCAAAAATA TTAAATATAC AAATTTCCGA 660

AACAATATAT TATGCCGCAC TAGAATCTAG TTGCGAACTA GCTAAAATTT ACGGACCTTA 720

TGAGACATAT AACGATTCTC CAGCGAGTAA AGGTATTCTA CAATATGATA TGTGGTTAAA 780

GAACCCAACA GATTTATGGG ATTGGAATGA ACTAAAAAAG AGAATTAATA CACATGGATT 840

GAGAAATAGC CTTCTAATAG CACCAATGCC TACTGCATCT ACATCTCAAA TATTAAGTAA 900

TAATGAGTCC ACCGAACCAT ATACTAGCAA TATATATACA AGAAGAGTAT TATCTGGAGA 960

TTTTCAGGTT GTAAATCCAC ACCTATTGAG AGAACTAATA AGTAGAAATA TGTGGAATAA 1020

TGACATAAAG AATACAATTG TGTTACATAA TGGTTCTATT CAACATTTAG ATTTACCAGA 1080

TAATATAAAA CCAATATATA AAACGGTTTG GGAGATATCT CCAAAATGTA TTTTAGAAAT 1140

GGCAGCCGAC AGAGGTGCGT TTATAGATCC AAGTCAATCA ATGACAATAT ATATAGATAA 1200

TCCTACATAC GCAAAACTGA CCAGTATGCA TTTTTACGGA TGGAGATTGG GGCTAAAAAC 1260

TGGGATGTAT TATATGAGAA CAAAATCGGC ATCAAATCCT ATAAAATTCA CAGTTGAGTG 1320

TAGTAATTGT TCTGCATAAT TTTTATAAAA ATGAAATACT ATCTCATGTA TCTTAATATA 1380

TTAAAAATGC GTAAAAGTGG CATTCCAAAA CAACCCGTTC CCAAAAAAGA TTATGTTCAA 1440

ACTGATAATA ATAAAAAACA ACAAATAACA ACGTGTTCAG AAGTCGTTGA GTATCTTAAA 1500

TCACTAAGTA AGAGCACCGA AAAATGTATA GAAAATGTAA TATTAACGCC TTCTCAATAT 1560

CCTTCTTGTT CATCGATAAC TATTAATTTA ACAGACTATC TATCATCTAA AATGACATCT 1620

ACATATATAG CATTAGAAGG TGAGTCTAAA ATATACAAGA ATAAAAAGAA TGAAAGTAGA 1680

TCGTTAGATC AATATTTTTT AAAAATACGA CTTACTGCAG CAAGTCCTAT AATGTATCAA 1740

TTATTAGATT GTATATATTC TAATATTAGA GATAATAAAC ATATACCCCC TTCCTTATCA 1800

AATATATCTA TATCGGACTT AGAAGAGAAA ACGCTTAACC AGGGGTGTTT GTTCATTAAT 1860

AAGATGGGTG GAGCTATTAT AGAATACAAG ATACCTGGTT CCAAATCTAT AACAAAATCT 1920

ATTTCCGAAG AACTAGAAAA TTTAACAAAG CGAGATAAAC AAATATCTAA AATTATAGTT 1980

ATTCCTATTG TATGTTACAG AAATGCAAAT AGTATAAAGG TTACATTTGC ACTAAAAAAG 2040

TTTATCATAG ATAAGGAGTT TAGTACAAAT GTAATAGACG TAGATGGTAA ACATGAAAAA 2100

ATGTCCATGA ATGAAACATG CGAAGAGGAT GTTGCTAGAG GATTGGGAAT TATAGATCTT 2160 GAAGATGAAT GCATAGAGGA AGATGATGTC GATACGTCAT TATTTAATGT ATAAATGGAT 2220

AAATTGTATG CGGCAATATT CGGCGTTTTT ATGACATCTA AAGATGATGA TTTTAATAAC 2280

TTTATAGAAG TTGTAAAATC TGTATTAACA GATACATCAT CTAATCATAC AATATCGTCG 2340

TCCAATAATA ATACATGGAT ATATATATTT CTAGCGATAT TATTTGGTGT TATGGTATTA 2400

TTAGTTTTTA TTTTGTATTT AAAAGTTACT AAACCAACTT AAATGGAGGA AGCAGATAAC 2460

CAACTCGTTT TAAATAGTAT TAGTGCTAGA GCATTAAAGG CATTTTTTGT ATCTAAAATT 2520

AATGATATGG TCGATGAATT AGTTACCAAA AAATATCCAC CAAAGAAGAA ATCACAAATA 2580

AAACTCATAG ATACACGAAT TCCTATTGAT CTTATTAATC AACAATTCGT TAAAAGATTT 2640

AAACTAGAAA ATTATAAAAA TGGAATTTTA TCCGTTCTTA TCAATAGTTT AGTCGAAAAT 2700

AATTACTTTG AACAAGATGG TAAACTTAAT AGCAGTGATA TTGATGAATT AGTGCTCACA 2760

GACATAGAGA AAAAGATTTT ATCGTTGATT CCTAGATGTT CTCCTCTTTA TATAGATATC 2820

AGTGACGTTA AAGTTCTCGC ATCTAGGTTA AAAAAGTGCT AAATCATTTA CGTTTAATGA 2880

TCATGAATAT ATTATACAAT CTGATAAAAT AGAGGAATTA ATAAATAGTT TATCTAGAAA 2940

CCATGATATT ATACTAGATG AAAAAAGTTC TATTAAAGAC AGCATATATA TACTATCTGA 3000

TGATCTTTTG AATATACTTC GTGAAAGATT ATTTAGATGT CCACAGGTTA AAGATAATAC 3060

TATTTCTAGA ACACGTCTAT ATGATTATTT TACTAGAGTG TCAAAGAAAG AAGAAGCGAA 3120

AATATACGTT ATATTGAAAG ATTTAAAGAT TGCTGATATA CTCGGTATCG AAACAGTAAC 3180

GATAGGATCA TTTGTATATA CGAAATATAG CATGTTGATT AATTCAATTT CGTCTAATGT 3240

TGATAGATAT TCAAAAAGGT TCCATGACTC TTTTTATGAA GATATTGCGG AATTTATAAA 3300

GGATAATGAA AAAATTAATG TATCCAGAGT TGTTGAATGC CTTATCGTAC CTAATATTAA 3360

TATAGAGTTA TTAACTGAAT AAGTATATAT AAATGATTGT TTTTATAATG TTTGTTATCG 3420

CATTTAGTTT TGCTGTATGG TTATCATATA CATTTTTAAG GCCGTATATG ATAAATGAAA 3480

ATATATAAGC ACTTATTTTT GTTAGTATAA TAACACAATG CCGTCGTATA TGTATCCGAA 3540

GAACGCAAGA AAAGTAATTT CAAAGATTAT ATCATTACAA CTTGATATTA AAAAACTTCC 3600

TAAAAAATAT ATAAATACCA TGTTAGAATT TGGTCTACAT GGAAATCTAC CAGCTTGTAT 3660

GTATAAAGAT GCCGTATCAT ATGATATAAA TAATATAAGA TTTTTACCTT ATAATTGTGT 3720

TATGGTTAAA GATTTAATAA ATGTTATAAA ATCATCATCT GTAATAGATA CTAGATTACA 3780

TCAATCTGTA TTAAAACATC GTAGAGCGTT AATAGATTAC GGCGATCAAG ACATTATCAC 3840

TTTAATGATC ATTAATAAGT TACTATCGAT AGATGATATA TCCTATATAT TAGATAAAAA 3900

AATAATTCAT GTAACAAAAA TATTAAAAAT AGACCCTACA GTAGCCAATT CAAACATGAA 3960

ACTGAATAAG ATAGAGCTTG TAGATGTAAT AACATCAATA CCTAAGTCTT CCTATACATA 4020

TTTATATAAT AATATGATCA TTGATCTCGA TACATTATTA TATTTATCCG ATGCATTCCA 4080

CATACCCCCC ACACATATAT CATTACGTTC ACTTAGAGAT ATAAACAGGA TTATTGAATT 4140

GCTTAAAAAA TATCCGAATA ATAATATTAT TGATTATATA TCCGATAGCA TAAAATCAAA 4200 TAGTTCATTC ATTCACATAC TTCATATGAT AATATCAAAT ATGTTTCCTG CTATAATCCC 4260

TAGTGTAAAC GATTTTATAT CTACCGTAGT TGATAAAGAT CGACTTATTA ATATGTATGG 4320

GATTAAGTGT GTTGCTATGT TTTCGTACGA TATAAACATG ATCGATTTAG AGTCATTAGA 4380

TGACTCAGAT TACATATTTA TAGAAAAAAA TATATCTATA TACGACGTTA AATGTAGAGA 4440

TTTTGCGAAT ATGATTAGAG ATAAGGTTAA AAGAGAAAAG AATAGAATAT TAACTACGAA 4500

ATGTGAAGAT ATTATAAGAT ATATAAAATT ATTCAGTAAA AATAGAATAA ACGATGAAAA 4560

TAATAAGGTG GAGGAGGTGT TGATACATAT TGATAATGTA TCTAAAAATA ATAAATTATC 4620

ACTGTCTGAT ATATCATCTT TAATGGATCA ATTTCGTTTA AATCCATGTA CCATAAGAAA 4680

TATATTATTA TCTTCAGCAA CTATAAAATC AAAACTATTA GCGTTACGGG CAGTAAAAAA 4740

CTGGAAATGT TATTCATTGA CAAATGTATC AATGTATAAA AAAATAAAGG GTGTTATCGT 4800

AATGGATATG GTTGATTATA TATCTACTAA CATTCTTAAA TACCATAAAC AATTATATGA 4860

TAAAATGAGT ACGTTTGAAT ATAAACGAGA TATTAAATCA TGTAAATGCT CGATATGTTC 4920

CGACTCTATA ACACATCATA TATATGAAAC AACATCATGT ATAAATTATA AATCTACCGA 4980

TAATGATCTT ATGATAGTAT TGTTCAATCT AACTAGATAT TTAATGCATG GGATGATACA 5040

TCCTAATCTT ATAAGCGTAA AAGGATGGGG TCCCCTTATT GGATTATTAA CGGGTGATAT 5100

AGGTATTAAT TTAAAACTAT ATTCCACCAT GAATATAAAT GGGCTACGGT ATGGAGATAT 5160

TACGTTATCT TCATACGATA TGAGTAATAA ATTAGTCTCT ATTATTAATA CACCCATATA 5220

TGAGTTAATA CCGTTTACTA CATGTTGTTC ACTCAATGAA TATTATTCAA AAATTGTGAT 5280

TTTAATAAAT GTTATTTTAG AATATATGAT ATCTATTATA TTATATAGAA TATTGATCGT 5340

AAAAAGATTT AATAACATTA AAGAATTTAT TTCAAAAGTC GTAAATACTG TACTAGAATC 5400

ATCAGGCATA TATTTTTGTC AGATGCGTGT ACATGAACAA ATTGAATTGG AAATAGATGA 5460

GCTCATTATT AATGGATCTA TGCCTGTACA GCTTATGCAT TTACTTCTAA AGGTAGCTAC 5520

CATAATATTA GAGGAAATCA AAGAAATATA ACGTATTTTT TCTTTTAAAT AAATAAAAAT 5580

ACTTTTTTTT TTAAACAAGG GGTGCTACCT TGTCTAATTG TATCTTGTAT TTTGGATCTG 5640

ATGCAAGATT ATTAAATAAT CGTATGAAAA AGTAGTAGAT ATAGTTTATA TCGTTACTGG 5700

ACATGATATT ATGTTTAGTT AATTCTTCTT TGGCATGAAT TCTACACGTC GGACAAGGTA 5760

ATGTATCTAT AATGGTATAA AGCTT 5785

(2) INFORMATION FOR SEQ ID NO:222:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 722 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: N ( iv) ANTI - SENSE : N

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:222:

TTTTGATTTT ACGCCATTAT ACTGTTCTGT AGATGCAAAT AATGAAGATG TGTTCTTATT 60

TACTAGAGAG ATGCAGACCC TATATTATCA CAGTATTTGG TGAACGTGTA TACTAACAGC 120

TTCAATAATC ATAATCCCCC ATATTATATA ACTATTAAAT TATGATATAG ATATAAATAC 180

TATCCAAAAT ACATTATTTA AACTGGAACA AGATATTATT AACTCTACCA TAGATACTTA 240

CTATTACAAT AATCTTGTTA AAAAAGAACA TTTTATAAAA TTATTTCTAG CCTACATAGT 300

TAAGAGGTAT GAAAAAAATA TAGGAATATT ATTTCTTGAT TATCCCACTC TTGGTGAATA 360

TTTCGTGAAA TTTATAGATA CGTGTATGAT GGAAATATTT GAGATGAAAT CAGATAAGGT 420

GGTAAACGGA TATATATTCT ATTATATTTA CGAATAAGTA TATTCCTATC CCATATATAA 480

CGTGTAAAAA GCTAAAGAAA TACGAATCCT TTGTTGTATA TGGAACCGAA ATAAAATCAA 540

TAATAAAATC TTCAAAGATT AGATATGCGA GTGTTATAAA AGTAACGGAG TATATCACAT 600

CTATCTGTTC GGAAGAAACT AGTTTATGGA ACAGCATCCC AATTGAGATA AAACATAAGA 660

TTATTAATAA TATAAACAAT CATGATATGT ATATATTATA TAAAAATAGA AAAAAAAAAT 720

AA 722

(2) INFORMATION FOR SEQ ID NO:223:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 234 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: N (iv) ANTI-SENSE: N

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:223:

AAACAATGCG CTTTAATATC AAACATGCAG GTGGAATAGG ATTGTCGATA AGTAATATAC 60

GAGCTAAGGG TACTTATATA TCCGGTATAA ACGGCAAATC TATGGTATAG TACCTATGTT 120

AAGAATATAT AATAACACAG TTAGATATAT TAATCAGGGA GGTGATAAAA GACCAGGAGC 180

AATGTCGATT TATATAGAAC CATGGCACGC TGATATATTC GATTTTCTAA GCTT 234 (2) INFORMATION FOR SEQ ID NO:224:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 1025 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: N (iv) ANTI-SENSE: N

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:224:

GGTTGCTCCT AACTTAATAA GATAATCCAC CAAGATAGTT TTATCCGTGG TAGATGCATA 60

CACAACAGGA GAATATCCTA ATTTATCTCT ATAGTTTATG GTTGTGATAT CTATAGTATA 120

TGGGACCGCC GAAAAACATG TATAATCGTC GTGACAATAG TTTAACATCG TGTTTAATAT 180

CGACATCATT TCATCATTTT TATTATATTC ATGTTTTATA TGCGAACAAA GCAAATTCAA 240

TATATTTAAA TTAGTGTTAT TGATGTGTCT AATTGTAAAT ATATGAATAG GATTCTTCAG 300

ACTATTATTT AGTTTACATA CATCAAATCC TTTTCTTATT AAAAACTCAA CAACTTTATA 360

ATCTATATTC TCATTACCAA GGTATTTATG CAATATGGTG TCTCCACATC TATGTACACT 420

GTTAATGTCA CCACCATGAT AAATAAGAAA CTTTATTACT TTAATTGTAA CATTCGTATT 480

AAATGTAAAA TAACAATGAA ATGGTGTTTT ATCATATATA GATATCCCAT TTAAATTAGC 540

ACCTTTATTA AGCAGTAATA ATACAATTTC TTTCAACTCT TTTAATTTAA ATACGTGCAA 600

CGATGAACTT AAAAATGTAG CTAACATATC AGTGGCTATA TTATCATCCT GTTTTATATT 660

TGATATTATT CTTCTTATAT TATCCATTTC CTTCTTACAA ACTATTTAAA CGATAACCAA 720

AATGTATTCA TGGGCTACTA ATAATAGCCA CATTACTAGA AAAAAAATTT TTTTTCAATA 780

TTATGACATT ATTACTTAAG TATTATTGAT AAGTCCTTCA TTGTTAAATG TAATAATATA 840

TATCGTTGTA TTTCTATAGG AATCCTCATC CAGTAACTAT GTTTCTTGCA GTGCTTCATA 900

ATTACATAAA TCGCTTTATC AATGTTAGAA TAATACATAT ATGTATTTTT GATAATATTT 960

TCTATATGTG ATCCATACAT TACTAAATTT TTTAATCTTA AAAAATTATC ATAATTGAGA 1020

AGCTT 1025 (2) INFORMATION FOR SEQ ID NO:225:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 305 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: N (iv) ANTI-SENSE: N

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:225: AAGCTTGGAT GAGCAATAAG AGTATACAAA ATTTAGTGTT TCAATTCGCT CATGGATCAG 60 AAGTAGAATA TATAGGTCAA TACGATATGA GATTTTTAAA TAATATACCT ATTCATGATA 120 AGTTTGATGT GTTTTTAAAT AAGCACATAC TATCGTATGT ACTTAGAGAT AAAATAAAGA 180

AATCAGACCA CAGATATGTA ATGTTTGGAT TTTGGTTATT TATCTCATTG GAAATGTGTT 240

ATATTCGATA AGGAACATCA TATGTCTGTT TCTATGATTC AGGAGGAATT ACCAAACGAA 300

TTCCA 305

(2) INFORMATION FOR SEQ ID NO:226:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 1721 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: N (iv) ANTI-SENSE: N

(ix) FEATURE:

(A) NAME/KEY: CDS

(B) LOCATION: 1..1721 (D) OTHER INFORMATION:

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:226:

ATG AAT TCG GAT CCG GCA ATA CTA TTA GTC TTG CTA TGT ACA TTT ACA 48 Met Asn Ser Asp Pro Ala He Leu Leu Val Leu Leu Cys Thr Phe Thr 1 5 10 15

ACC GCA AAT GCA GAC ACA TTA TGT ATA GGT TAC CAT GCA AAT AAT TCA 96 Thr Ala Asn Ala Asp Thr Leu Cys He Gly Tyr His Ala Asn Asn Ser 20 25 30

ACT GAC ACT GTT GAC ACA GTA CTA GAA AAG AAT GTA ACA GTA ACA CAC 144 Thr Asp Thr Val Asp Thr Val Leu Glu Lys Asn Val Thr Val Thr His 35 40 45

TCT GTT AAC CTT CTA GAA GAC AGA CAT AAC GGG AAA CTA TGT AAA CTA 192 Ser Val Asn Leu Leu Glu Asp Arg His Asn Gly Lys Leu Cys Lys Leu 50 55 60

AGA GGG GTA GCC CCA TTG CAT TTG GGT AAA TGT AAC ATT GCT GGA TGG 240 Arg Gly Val Ala Pro Leu His Leu Gly Lys Cys Asn He Ala Gly Trp 65 70 75 80

CTC CTG GGA AAC CCA GAG TGT GAA TTA CTA TTC ACA GCA AGC TCA TGG 288 Leu Leu Gly Asn Pro Glu Cys Glu Leu Leu Phe Thr Ala Ser Ser Trp 85 90 95

TCT TAC ATT GTG GAA ACA TCT AAT TCA GAC AAT GGG ACA TGT TAC CCA 336 Ser Tyr He Val Glu Thr Ser Asn Ser Asp Asn Gly Thr Cys Tyr Pro 100 105 110

GGA GAT TTC ATC AAT TAT GAA GAG CTA AGA GAG CAG TTG AGC TCA GTG 384 Gly Asp Phe He Asn Tyr Glu Glu Leu Arg Glu Gin Leu Ser Ser Val 115 120 125

TCA TCA TTT GAA AGA TTT GAG ATA TTC CCC AAG GCA AGT TCA TGG CCC 432 Ser Ser Phe Glu Arg Phe Glu He Phe Pro Lys Ala Ser Ser Trp Pro 130 135 140 AAT CAT GAA ACG AAC ATA GGT GTG ACG GCA GCA TGT CCT TAT GCT GGA 480 Asn His Glu Thr Asn He Gly Val Thr Ala Ala Cys Pro Tyr Ala Gly 145 150 155 160

GCA AAC AGC TTC TAC AGA AAC TTA ATA TGG CTG GTA AAA AAA GGA AAT 528 Ala Asn Ser Phe Tyr Arg Asn Leu He Trp Leu Val Lys Lys Gly Asn 165 170 175

TCA TAC CCA AAG CTC AGC AAA TCC TAT ATT AAC AAT AAG GAG AAG GAA 576 Ser Tyr Pro Lys Leu Ser Lys Ser Tyr He Asn Asn Lys Glu Lys Glu 180 185 190

GTC CTC GTG CTA TGG GGC ATT CAC CAT CCA CCT ACC AGT ACT GAC CAA 624 Val Leu Val Leu Trp Gly He His His Pro Pro Thr Ser Thr Asp Gin 195 200 205

CAA AGT CTC TAC CAG AAT GCA GAT GCC TAT GTT TTT GTG GGG TCA TCA 672 Gin Ser Leu Tyr Gin Asn Ala Asp Ala Tyr Val Phe Val Gly Ser Ser 210 215 220

AAA TAC AAC AAG AAA TTC AAG CCA GAA ATA GCA ACA AGA CCC AAG GTG 720 Lys Tyr Asn Lys Lys Phe Lys Pro Glu He Ala Thr Arg Pro Lys Val 225 230 235 240

AGA GGT CAA GCA GGG AGA ATG AAC TAT TAC TGG ACG CTA GTA AAG CCT 768 Arg Gly Gin Ala Gly Arg Met Asn Tyr Tyr Trp Thr Leu Val Lys Pro 245 250 255

GGA GAC ACA ATA ACA TTC GAA GCA ACT GGA AAT CTA GTG GTA CCA AGA 816 Gly Asp Thr He Thr Phe Glu Ala Thr Gly Asn Leu Val Val Pro Arg 260 265 270

TAT GCC TTC GCA ATG AAA AGA GGT TCT GGA TCT GGT ATT ATC ATT TCA 864 Tyr Ala Phe Ala Met Lys Arg Gly Ser Gly Ser Gly He He He Ser 275 280 285

GAT ACA CCA GTC CAC GAT TGT AAT ACG ACT TGT CAA ACA CCC AAA GGT 912 Asp Thr Pro Val His Asp Cys Asn Thr Thr Cys Gin Thr Pro Lys Gly 290 295 300

GCT ATA AAC ACC AGC CTT CCA TTT CAG AAT ATA CAT CCA GTC ACA ATT 960 Ala He Asn Thr Ser Leu Pro Phe Gin Asn He His Pro Val Thr He 305 310 315 320

GGA GAA TGT CCA AAA TAT GTC AAA AGC ACA AAA TTG AGA ATG GCT ACA 1008 Gly Glu Cys Pro Lys Tyr Val Lys Ser Thr Lys Leu Arg Met Ala Thr 325 330 335

GGA TTA AGG AAT ATC CCG TCT ATT CAA TCT AGA GGC CTG TTT GGA GCC 1056 Gly Leu Arg Asn He Pro Ser He Gin Ser Arg Gly Leu Phe Gly Ala 340 345 350

ATT GCT GGC TTT ATT GAG GGG GGA TGG ACA GGA ATG ATA GAT GGC TGG 1104 He Ala Gly Phe He Glu Gly Gly Trp Thr Gly Met He Asp Gly Trp 355 360 365

TAC GGT TAT CAC CAT CAG AAT GAG CAG GGA TCA GGA TAT GCA GCC GAC 1152 Tyr Gly Tyr His His Gin Asn Glu Gin Gly Ser Gly Tyr Ala Ala Asp 370 375 380

CGA AAG AGC ACA CAG AAT GCC ATT GAC GGG ATC ACT AAC AAA GTA AAC 1200 Arg Lys Ser Thr Gin Asn Ala He Asp Gly He Thr Asn Lys Val Asn 385 390 395 400

TCT GTT ATT GAA AAG ATG AAC ACA CAA TTC ACA GCA GTG GGT AAA GAA 1248 Ser Val He Glu Lys Met Asn Thr Gin Phe Thr Ala Val Gly Lys Glu 405 410 415 TTC AAC CAC CTG GAA AAA AGA ATA GAG AAT TTA AAC AAA AAG GTT GAT 1296 Phe Asn His Leu Glu Lys Arg He Glu Asn Leu Asn Lys Lys Val Asp 420 425 430

GAT GGT TTT CTG GAT GTT TGG ACT TAC AAT GCC GAA CTG TTG GTT CTA 1344 Asp Gly Phe Leu Asp Val Trp Thr Tyr Asn Ala Glu Leu Leu Val Leu 435 440 445

TTG GAA AAT GAA AGA ACT TTG GAT TAT CAC GAT TCA AAT GTG AAG AAC 1392 Leu Glu Asn Glu Arg Thr Leu Asp Tyr His Asp Ser Asn Val Lys Asn 450 455 460

CTA TAT GAG AAA GTA AGA AGC CAG CTA AAA AAC AAT GCC AAG GAA ATT 1440 Leu Tyr Glu Lys Val Arg Ser Gin Leu Lys Asn Asn Ala Lys Glu He 465 470 475 480

GGA AAT GGC TGC TTT GAA TTT TAC CAC AAA TGT GAT GAC ACG TGC ATG 1488 Gly Asn Gly Cys Phe Glu Phe Tyr His Lys Cys Asp Asp Thr Cys Met 485 490 495

GAG AGC GTC AAA AAT GGG ACT TAT GAT TAC CCA AAA TAC TCA GAG GAA 1536 Glu Ser Val Lys Asn Gly Thr Tyr Asp Tyr Pro Lys Tyr Ser Glu Glu 500 505 510

GCA AAA CTA AAC AGA GAG GAG ATA GAT GGG GTA AAG CTG GAA TCA ACA 1584 Ala Lys Leu Asn Arg Glu Glu He Asp Gly Val Lys Leu Glu Ser Thr 515 520 525

AGG ATT TAC CAG ATT TTG GCG ATC TAT TCA ACT GTC GCC AGT TCA TTG 1632 Arg He Tyr Gin He Leu Ala He Tyr Ser Thr Val Ala Ser Ser Leu 530 535 540

GTA CTG TTA GTC TCC CTG GGG GCA ATC AGT TTC TGG ATG TGC TCC AAT 1680 Val Leu Leu Val Ser Leu Gly Ala He Ser Phe Trp Met Cys Ser Asn 545 550 555 560

GGG TCT TTA CAG TGC AGA ATA TGT ATT TAA AAT TAG GAT CC 1721

Gly Ser Leu Gin Cys Arg He Cys He . Asn Asp 565 570

(2) INFORMATION FOR SEQ ID NO:227:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 573 amino acids

(B) TYPE: amino acid (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:227:

Met Asn Ser Asp Pro Ala He Leu Leu Val Leu Leu Cys Thr Phe Thr 1 5 10 15

Thr Ala Asn Ala Asp Thr Leu Cys He Gly Tyr His Ala Asn Asn Ser 20 25 30

Thr Asp Thr Val Asp Thr Val Leu Glu Lys Asn Val Thr Val Thr His 35 40 45

Ser Val Asn Leu Leu Glu Asp Arg His Asn Gly Lys Leu Cys Lys Leu 50 55 60

Arg Gly Val Ala Pro Leu His Leu Gly Lys Cys Asn He Ala Gly Trp 65 70 75 80 Leu Leu Gly Asn Pro Glu Cys Glu Leu Leu Phe Thr Ala Ser Ser Trp 85 90 95

Ser Tyr He Val Glu Thr Ser Asn Ser Asp Asn Gly Thr Cys Tyr Pro 100 105 110

Gly Asp Phe He Asn Tyr Glu Glu Leu Arg Glu Gin Leu Ser Ser Val 115 120 125

Ser Ser Phe Glu Arg Phe Glu He Phe Pro Lys Ala Ser Ser Trp Pro 130 135 140

Asn His Glu Thr Asn He Gly Val Thr Ala Ala Cys Pro Tyr Ala Gly 145 150 155 160

Ala Asn Ser Phe Tyr Arg Asn Leu He Trp Leu Val Lys Lys Gly Asn 165 170 175

Ser Tyr Pro Lys Leu Ser Lys Ser Tyr He Asn Asn Lys Glu Lys Glu 180 185 190

Val Leu Val Leu Trp Gly He His His Pro Pro Thr Ser Thr Asp Gin 195 200 205

Gin Ser Leu Tyr Gin Asn Ala Asp Ala Tyr Val Phe Val Gly Ser Ser 210 215 220

Lys Tyr Asn Lys Lys Phe Lys Pro Glu He Ala Thr Arg Pro Lys Val 225 230 235 240

Arg Gly Gin Ala Gly Arg Met Asn Tyr Tyr Trp Thr Leu Val Lys Pro 245 250 255

Gly Asp Thr He Thr Phe Glu Ala Thr Gly Asn Leu Val Val Pro Arg 260 265 270

Tyr Ala Phe Ala Met Lys Arg Gly Ser Gly Ser Gly He He He Ser 275 280 285

Asp Thr Pro Val His Asp Cys Asn Thr Thr Cys Gin Thr Pro Lys Gly 290 295 300

Ala He Asn Thr Ser Leu Pro Phe Gin Asn He His Pro Val Thr He 305 310 315 320

Gly Glu Cys Pro Lys Tyr Val Lys Ser Thr Lys Leu Arg Met Ala Thr 325 330 335

Gly Leu Arg Asn He Pro Ser He Gin Ser Arg Gly Leu Phe Gly Ala 340 345 350

He Ala Gly Phe He Glu Gly Gly Trp Thr Gly Met He Asp Gly Trp 355 360 365

Tyr Gly Tyr His His Gin Asn Glu Gin Gly Ser Gly Tyr Ala Ala Asp 370 375 380

Arg Lys Ser Thr Gin Asn Ala He Asp Gly He Thr Asn Lys Val Asn 385 390 395 400

Ser Val He Glu Lys Met Asn Thr Gin Phe Thr Ala Val Gly Lys Glu 405 410 415

Phe Asn His Leu Glu Lys Arg He Glu Asn Leu Asn Lys Lys Val Asp 420 425 430

Asp Gly Phe Leu Asp Val Trp Thr Tyr Asn Ala Glu Leu Leu Val Leu 435 440 445

Leu Glu Asn Glu Arg Thr Leu Asp Tyr His Asp Ser Asn Val Lys Asn 450 455 460

Leu Tyr Glu Lys Val Arg Ser Gin Leu Lys Asn Asn Ala Lys Glu He 465 470 475 480

Gly Asn Gly Cys Phe Glu Phe Tyr His Lys Cys Asp Asp Thr Cys Met 485 490 495

Glu Ser Val Lys Asn Gly Thr Tyr Asp Tyr Pro Lys Tyr Ser Glu Glu 500 505 510

Ala Lys Leu Asn Arg Glu Glu He Asp Gly Val Lys Leu Glu Ser Thr 515 520 525

Arg He Tyr Gin He Leu Ala He Tyr Ser Thr Val Ala Ser Ser Leu 530 535 540

Val Leu Leu Val Ser Leu Gly Ala He Ser Phe Trp Met Cys Ser Asn 545 550 555 560

Gly Ser Leu Gin Cys Arg He Cys He Asn Asp 565 570

(2) INFORMATION FOR SEQ ID NO:228:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 1414 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: N (iv) ANTI-SENSE: N

(ix) FEATURE:

(A) NAME/KEY: CDS

(B) LOCATION: 1..1414 (D) OTHER INFORMATION:

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:228:

ATG AAT TCA AAT CAA AAA ATA ATA ACC ATT GGG TCA ATC TGT CTG ATA 48 Met Asn Ser Asn Gin Lys He He Thr He Gly Ser He Cys Leu He 1 5 10 15

GTT GGA ATA GTT AGT CTA TTA TTG CAG ATA GGA AAT ATA GTC TCG TTA 96 Val Gly He Val Ser Leu Leu Leu Gin He Gly Asn He Val Ser Leu 20 25 30

TGG ATA AGC CAT TCA ATT CAG ACT GGA GAA AAA AAC CAC TCT GAG ATA 144 Trp He Ser His Ser He Gin Thr Gly Glu Lys Asn His Ser Glu He 35 40 45

TGC AAC CAA AAT ATC ATT ACA TAT GAA AAC AAC ACA TGG GTG AAC CAA 192 Cys Asn Gin Asn He He Thr Tyr Glu Asn Asn Thr Trp Val Asn Gin 50 55 60

ACT TAT GTA AAC ATT AGC AAT ACC AAC ATT GCT GAT GGA CAG GGC GTG 240 Thr Tyr Val Asn He Ser Asn Thr Asn He Ala Asp Gly Gin Gly Val 65 70 75 80

ACT TCA ATA ATA CTA GCC GGC AAT CCC CCT CTT TGC CCA ATA ATT GGG 288 Thr Ser He He Leu Ala Gly Asn Pro Pro Leu Cys Pro He He Gly 85 90 95

TGG GCT ATA TAC AGC AAA AAC AAT AGC ATA AGG ATT GGT CCC AAA GGA 336 Trp Ala He Tyr Ser Lys Asn Asn Ser He Arg He Gly Pro Lys Gly 100 105 110

AAC ATT TTT GTC ATA AAA AAA CCA TCC ATT TCA TGC TCT CAC TTG GAG 384 Asn He Phe Val He Lys Lys Pro Ser He Ser Cys Ser His Leu Glu 115 120 125

TGC AAA ACC TTT TTC CTG ACC CAA GGT GCT TTG CTA AAT GAC AGG CAT 432 Cys Lys Thr Phe Phe Leu Thr Gin Gly Ala Leu Leu Asn Asp Arg His 130 135 140

CCT AAT GGA ACC GTC AAG GAC AGG AGC CCT TAC CGA ACC TTA ATG AGC 480 Pro Asn Gly Thr Val Lys Asp Arg Ser Pro Tyr Arg Thr Leu Met Ser 145 150 155 160

TGC CCG ATC GGT GAA GCT CCA TCT CCG TAT AAT TCA AGA TTC GAA TCA 528 Cys Pro He Gly Glu Ala Pro Ser Pro Tyr Asn Ser Arg Phe Glu Ser 165 170 175

GTT GCT TGG TCA GCA AGT GCA TGC CAT GAT GGA ATG GGA TGG CTA ACA 576 Val Ala Trp Ser Ala Ser Ala Cys His Asp Gly Met Gly Trp Leu Thr 180 185 190

ATC GGG ATT TCC GGT CCA GAT AAT GGA GCA GTG GCT GTT TTG AAA TAC 624 He Gly He Ser Gly Pro Asp Asn Gly Ala Val Ala Val Leu Lys Tyr 195 200 205

AAT GGT ATA ATA ACA GAT ACA ATA AAA AGT TGG AGA AAC AAA ATA CTA 672 Asn Gly He He Thr Asp Thr He Lys Ser Trp Arg Asn Lys He Leu 210 215 220

AGA ACA CAA GAG TCA GAA TGT GTT TGT ATA AAC GGT TCA TGT TTT ACT 720 Arg Thr Gin Glu Ser Glu Cys Val Cys He Asn Gly Ser Cys Phe Thr 225 230 235 240

ATA ATG ACT GAT GGC CCA AGC AAT GGG CAA GCC TCG TAC AAA ATA TTC 768 He Met Thr Asp Gly Pro Ser Asn Gly Gin Ala Ser Tyr Lys He Phe 245 250 255

AAA ATG GAG AAA GGG AAG ATT ATT AAG TCA GTT GAG CTG GAT GCA CCT 816 Lys Met Glu Lys Gly Lys He He Lys Ser Val Glu Leu Asp Ala Pro 260 265 270

AAT TAC CAC TAT GAG GAA TGC TCC TGT TAC CCT GAT ACA GGC AAA GTG 864 Asn Tyr His Tyr Glu Glu Cys Ser Cys Tyr Pro Asp Thr Gly Lys Val 275 280 285

GTG TGT GTG TGC AGA GAC AAT TGG CAT GCT TCA AAT CGA CCG TGG GTC 912 Val Cys Val Cys Arg Asp Asn Trp His Ala Ser Asn Arg Pro Trp Val 290 295 300

TCT TTC GAT CAG AAT CTT GAT TAT CAG ATA GGG TAC ATA TGC AGT GGG 960 Ser Phe Asp Gin Asn Leu Asp Tyr Gin He Gly Tyr He Cys Ser Gly 305 310 315 320

GTT TTC GGT GAT AAT CCG CGT TCT AAT GAT GGG AAA GGC AAT TGT GGC 1008 Val Phe Gly Asp Asn Pro Arg Ser Asn Asp Gly Lys Gly Asn Cys Gly 325 330 335

CCA GTA CTT TCT AAT GGA GCA AAT GGA GTG AAA GGA TTC TCA TTT AGA 1056 Pro Val Leu Ser Asn Gly Ala Asn Gly Val Lys Gly Phe Ser Phe Arg 340 345 350

TAT GGC AAT GGT GTT TGG ATA GGA AGA ACT AAA AGT ATC AGC TCT AGA 1104 Tyr Gly Asn Gly Val Trp He Gly Arg Thr Lys Ser He Ser Ser Arg 355 360 365

AGT GGA TTT GAG ATG ATT TGG GAT CCA AAT GGA TGG ACG GAA ACT GAT 1152 Ser Gly Phe Glu Met He Trp Asp Pro Asn Gly Trp Thr Glu Thr Asp 370 375 380

AGT AGT TTC TCT ATA AAG CAG GAT ATT ATA GCA TTA ACT GAT TGG TCA 1200 Ser Ser Phe Ser He Lys Gin Asp He He Ala Leu Thr Asp Trp Ser 385 390 395 400

GGA TAC AGT GGA AGT TTT GTC CAA CAT CCT GAA TTA ACA GGA ATG AAC 1248 Gly Tyr Ser Gly Ser Phe Val Gin His Pro Glu Leu Thr Gly Met Asn 405 410 415

TGC ATA AGG CCT TGT TTT TGG GTA GAG TTA ATC AGA GGA CAA CCC AAG 1296 Cys He Arg Pro Cys Phe Trp Val Glu Leu He Arg Gly Gin Pro Lys 420 425 430

GAG AGC ACA ATC TGG ACT AGT GGA AGC AGC ATT TCT TTC TGT GGC GTG 1344 Glu Ser Thr He Trp Thr Ser Gly Ser Ser He Ser Phe Cys Gly Val 435 440 445

GAC AAT GAA ACC GCA AGC TGG TCA TGG CCA GAC GGA GCT GAT CTG CCA 1392 Asp Asn Glu Thr Ala Ser Trp Ser Trp Pro Asp Gly Ala Asp Leu Pro 450 455 460

TTC ACC ATT GAC AAG TAG ATC T 1414

Phe Thr He Asp Lys He 465 470

(2) INFORMATION FOR SEQ ID NO:229:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 471 amino acids

(B) TYPE: amino acid (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:229:

Met Asn Ser Asn Gin Lys He He Thr He Gly Ser He Cys Leu He 1 5 10 15

Val Gly He Val Ser Leu Leu Leu Gin He Gly Asn He Val Ser Leu 20 25 30

Trp He Ser His Ser He Gin Thr Gly Glu Lys Asn His Ser Glu He 35 40 45

Cys Asn Gin Asn He He Thr Tyr Glu Asn Asn Thr Trp Val Asn Gin 50 55 60

Thr Tyr Val Asn He Ser Asn Thr Asn He Ala Asp Gly Gin Gly Val 65 70 75 80

Thr Ser He He Leu Ala Gly Asn Pro Pro Leu Cys Pro He He Gly 85 90 95

Trp Ala He Tyr Ser Lys Asn Asn Ser He Arg He Gly Pro Lys Gly 100 105 110 Asn He Phe Val He Lys Lys Pro Ser He Ser Cys Ser His Leu Glu 115 120 125

Cys Lys Thr Phe Phe Leu Thr Gin Gly Ala Leu Leu Asn Asp Arg His 130 135 140

Pro Asn Gly Thr Val Lys Asp Arg Ser Pro Tyr Arg Thr Leu Met Ser 145 150 155 160

Cys Pro He Gly Glu Ala Pro Ser Pro Tyr Asn Ser Arg Phe Glu Ser 165 170 175

Val Ala Trp Ser Ala Ser Ala Cys His Asp Gly Met Gly Trp Leu Thr 180 185 190

He Gly He Ser Gly Pro Asp Asn Gly Ala Val Ala Val Leu Lys Tyr 195 200 205

Asn Gly He He Thr Asp Thr He Lys Ser Trp Arg Asn Lys He Leu 210 215 220

Arg Thr Gin Glu Ser Glu Cys Val Cys He Asn Gly Ser Cys Phe Thr 225 230 235 240

He Met Thr Asp Gly Pro Ser Asn Gly Gin Ala Ser Tyr Lys He Phe 245 250 255

Lys Met Glu Lys Gly Lys He He Lys Ser Val Glu Leu Asp Ala Pro 260 265 270

Asn Tyr His Tyr Glu Glu Cys Ser Cys Tyr Pro Asp Thr Gly Lys Val 275 280 285

Val Cys Val Cys Arg Asp Asn Trp His Ala Ser Asn Arg Pro Trp Val 290 295 300

Ser Phe Asp Gin Asn Leu Asp Tyr Gin He Gly Tyr He Cys Ser Gly 305 310 315 320

Val Phe Gly Asp Asn Pro Arg Ser Asn Asp Gly Lys Gly Asn Cys Gly 325 330 335

Pro Val Leu Ser Asn Gly Ala Asn Gly Val Lys Gly Phe Ser Phe Arg 340 345 350

Tyr Gly Asn Gly Val Trp He Gly Arg Thr Lys Ser He Ser Ser Arg 355 360 365

Ser Gly Phe Glu Met He Trp Asp Pro Asn Gly Trp Thr Glu Thr Asp 370 375 380

Ser Ser Phe Ser He Lys Gin Asp He He Ala Leu Thr Asp Trp Ser 385 390 395 400

Gly Tyr Ser Gly Ser Phe Val Gin His Pro Glu Leu Thr Gly Met Asn 405 410 415

Cys He Arg Pro Cys Phe Trp Val Glu Leu He Arg Gly Gin Pro Lys 420 425 430

Glu Ser Thr He Trp Thr Ser Gly Ser Ser He Ser Phe Cys Gly Val 435 440 445

Asp Asn Glu Thr Ala Ser Trp Ser Trp Pro Asp Gly Ala Asp Leu Pro 450 455 460

Phe Thr He Asp Lys . He 465 470

(2) INFORMATION FOR SEQ ID NO:230:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 1501 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: N (iv) ANTI-SENSE: N

(ix) FEATURE:

(A) NAME/KEY: CDS

(B) LOCATION: 1..1501 (D) OTHER INFORMATION:

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:230:

ATG AAT TCT CAA GGC ACC AAA CGA TCA TAT GAA CAA ATG GAG ACT GGT 48 Met Asn Ser Gin Gly Thr Lys Arg Ser Tyr Glu Gin Met Glu Thr Gly 1 5 10 • 15

GGG GAA CGC CAG GAT GCC ACA GAA ATC AGA GCA TCT GTC GGA AGA ATG 96 Gly Glu Arg Gin Asp Ala Thr Glu He Arg Ala Ser Val Gly Arg Met 20 25 30

ATT GGT GGA ATC GGA AGA TTC TAC ATC CAA ATG TGC ACT GAA CTC AAA 144 He Gly Gly He Gly Arg Phe Tyr He Gin Met Cys Thr Glu Leu Lys 35 40 45

CTC AGT GAC TAT GAG GGA CGA CTA ATT CAA AAT AGC ATA ACA ATA GAG 192 Leu Ser Asp Tyr Glu Gly Arg Leu He Gin Asn Ser He Thr He Glu 50 55 60

AGA ATG GTG CTC TCT GCT TTT GAT GAG AGA AGG AAT AAA TAC CTA GAA 240 Arg Met Val Leu Ser Ala Phe Asp Glu Arg Arg Asn Lys Tyr Leu Glu 65 70 75 80

GAG CAT CCC AGT GCT GGG AAG GAT CCT AAG AAA ACT GGA GGA CCC ATA 288 Glu His Pro Ser Ala Gly Lys Asp Pro Lys Lys Thr Gly Gly Pro He 85 90 95

TAT AGA AGG GTA GAC GGA AAA TGG ATG AGA GAA CTC ATC CTT TAT GAC 336 Tyr Arg Arg Val Asp Gly Lys Trp Met Arg Glu Leu He Leu Tyr Asp 100 105 110

AAA GAA GAA ATA AGG AGA GTT TGG CGC CAA GCA AAC AAT GGT GAG GAT 384 Lys Glu Glu He Arg Arg Val Trp Arg Gin Ala Asn Asn Gly Glu Asp 115 120 125

GCA ACA GCC GGT CTT ACT CAC ATC ATG ATT TGG CAC TCC AAT CTT AAT 432 Ala Thr Ala Gly Leu Thr His He Met He Trp His Ser Asn Leu Asn 130 135 140

GAT GCC ACC TAT CAG AGA ACA AGA GCG CTT GTT CGC ACT GGA ATG GAT 480 Asp Ala Thr Tyr Gin Arg Thr Arg Ala Leu Val Arg Thr Gly Met Asp 145 150 155 160

CCC AGA ATG TGC TCC CTA ATG CAA GGT TCA ACA CTT CCC AGA AGG TCT 528 Pro Arg Met Cys Ser Leu Met Gin Gly Ser Thr Leu Pro Arg Arg Ser 165 170 175

GGG GCC GCA GGT GCT GCA GTG AAA GGA GTT GGA ACA ATA GCA ATG GAG 576 Gly Ala Ala Gly Ala Ala Val Lys Gly Val Gly Thr He Ala Met Glu 180 185 190

TTA ATC AGA ATG ATC AAA CGT GGA ATC AAT GAC CGA AAC TTC TGG AGG 624 Leu He Arg Met He Lys Arg Gly He Asn Asp Arg Asn Phe Trp Arg 195 200 205

GGT GAA AAT GGA CGA AGG ACA AGG ATT GCA TAT GAA AGA ATG TGC AAT 672 Gly Glu Asn Gly Arg Arg Thr Arg He Ala Tyr Glu Arg Met Cys Asn 210 215 220

ATT CTC AAA GGA AAA TTT CAG ACA GCT GCC CAG AGG GCA ATG ATG GAT 720 He Leu Lys Gly Lys Phe Gin Thr Ala Ala Gin Arg Ala Met Met Asp 225 230 235 240

CAA GTA AGA GAA AGT CGA AAC CCA GGA AAC GCT GAA ATT GAA GAT CTC 768 Gin Val Arg Glu Ser Arg Asn Pro Gly Asn Ala Glu He Glu Asp Leu 245 250 255

ATT TTC CTG GCA CGG TCA GCA CTT ATT CTA AGG GGG TCA GTT GCA CAT 816 He Phe Leu Ala Arg Ser Ala Leu He Leu Arg Gly Ser Val Ala His 260 265 270

AAG TCC TGC CTG CCT GCT TGT GTG TAT GGG CTT GCA GTA GCA AGT GGG 864 Lys Ser Cys Leu Pro Ala Cys Val Tyr Gly Leu Ala Val Ala Ser Gly 275 280 285

CAT GAC TTT GAA AGA GAA GGA TAT TCA CTG GTC GGG ATA GAC CCC TTC 912 His Asp Phe Glu Arg Glu Gly Tyr Ser Leu Val Gly He Asp Pro Phe 290 295 300

AAA TTA CTT CAA AAC AGT CAA GTG TTC AGC CTG ATC AGA CCA AAT GAA 960 Lys Leu Leu Gin Asn Ser Gin Val Phe Ser Leu He Arg Pro Asn Glu 305 310 315 320

AAC CCA GCT CAC AAG AGT CAA TTG GTG TGG ATG GCA TGC CAT TCT GCT 1008 Asn Pro Ala His Lys Ser Gin Leu Val Trp Met Ala Cys His Ser Ala 325 330 335

GCA TTT GAG GAT TTA AGA ATA TCA AGT TTC ATA AGA GGG AAG AAA GTG 1056 Ala Phe Glu Asp Leu Arg He Ser Ser Phe He Arg Gly Lys Lys Val 340 345 350

GTT CCA AGA GGA AAG CTT TCC ACA AGA GGG GTT CAG ATT GCT TCA AAT 1104 Val Pro Arg Gly Lys Leu Ser Thr Arg Gly Val Gin He Ala Ser Asn 355 360 365

GAG AAT GTG GAA GCT ATG GAC TCT AGT ACC CTA AAA CTA AGA AGC AGA 1152 Glu Asn Val Glu Ala Met Asp Ser Ser Thr Leu Lys Leu Arg Ser Arg 370 375 380

TAT TGG GCC ATA AGG ACC AGA AGT GGA GGA AAT ACC AAC CAA CAG AAG 1200 Tyr Trp Ala He Arg Thr Arg Ser Gly Gly Asn Thr Asn Gin Gin Lys 385 390 395 400

GCA TCT GCG GGC CAG ATC AGT GTG CAA CCT ACA TTC TCA GTG CAA CGG 1248 Ala Ser Ala Gly Gin He Ser Val Gin Pro Thr Phe Ser Val Gin Arg 405 410 415

AAT CTC CCT TTT GAA AGA GCA ACC GTT ATG GCA GCT TTC AGC GGG AAT 1296 Asn Leu Pro Phe Glu Arg Ala Thr Val Met Ala Ala Phe Ser Gly Asn 420 425 430

AAT GAG GGA CGG ACA TCA GAC ATG CGA ACG GAA GTT ATA AGG ATG ATG 1344 Asn Glu Gly Arg Thr Ser Asp Met Arg Thr Glu Val He Arg Met Met 435 440 445

GAA AGT GCA AAG CCA GAA GAT TTG TCC TTC CAG GGG CGG GGA GTC TTC 1392 Glu Ser Ala Lys Pro Glu Asp Leu Ser Phe Gin Gly Arg Gly Val Phe 450 455 460

GAG CTC TCG GAC GAA AAG GCA ACG AAC CCG ATC GTG CCT TCC TTT GAC 1440 Glu Leu Ser Asp Glu Lys Ala Thr Asn Pro He Val Pro Ser Phe Asp 465 470 475 480

ATG AGT AAT GAA GGG TCT TAT TTC TTC GGA GAC AAT GCA GAG GAG TAT 1488 Met Ser Asn Glu Gly Ser Tyr Phe Phe Gly Asp Asn Ala Glu Glu Tyr 485 490 495

GAC AAT TGA ATT C 1501

Asp Asn He 500

(2) INFORMATION FOR SEQ ID NO:231:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 500 amino acids

(B) TYPE: amino acid (D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:231:

Met Asn Ser Gin Gly Thr Lys Arg Ser Tyr Glu Gin Met Glu Thr Gly 1 5 10 15

Gly Glu Arg Gin Asp Ala Thr Glu He Arg Ala Ser Val Gly Arg Met 20 25 30

He Gly Gly He Gly Arg Phe Tyr He Gin Met Cys Thr Glu Leu Lys 35 40 45

Leu Ser Asp Tyr Glu Gly Arg Leu He Gin Asn Ser He Thr He Glu 50 55 60

Arg Met Val Leu Ser Ala Phe Asp Glu Arg Arg Asn Lys Tyr Leu Glu 65 70 75 80

Glu His Pro Ser Ala Gly Lys Asp Pro Lys Lys Thr Gly Gly Pro He 85 90 95

Tyr Arg Arg Val Asp Gly Lys Trp Met Arg Glu Leu He Leu Tyr Asp 100 105 110

Lys Glu Glu He Arg Arg Val Trp Arg Gin Ala Asn Asn Gly Glu Asp 115 120 125

Ala Thr Ala Gly Leu Thr His He Met He Trp His Ser Asn Leu Asn 130 135 140

Asp Ala Thr Tyr Gin Arg Thr Arg Ala Leu Val Arg Thr Gly Met Asp 145 150 155 160

Pro Arg Met Cys Ser Leu Met Gin Gly Ser Thr Leu Pro Arg Arg Ser 165 170 175

Gly Ala Ala Gly Ala Ala Val Lys Gly Val Gly Thr He Ala Met Glu 180 185 190 Leu He Arg Met He Lys Arg Gly He Asn Asp Arg Asn Phe Trp Arg 195 200 205

Gly Glu Asn Gly Arg Arg Thr Arg He Ala Tyr Glu Arg Met Cys Asn 210 215 220

He Leu Lys Gly Lys Phe Gin Thr Ala Ala Gin Arg Ala Met Met Asp 225 230 235 240

Gin Val Arg Glu Ser Arg Asn Pro Gly Asn Ala Glu He Glu Asp Leu 245 250 255

He Phe Leu Ala Arg Ser Ala Leu He Leu Arg Gly Ser Val Ala His 260 265 270

Lys Ser Cys Leu Pro Ala Cys Val Tyr Gly Leu Ala Val Ala Ser Gly 275 280 285

His Asp Phe Glu Arg Glu Gly Tyr Ser Leu Val Gly He Asp Pro Phe 290 295 300

Lys Leu Leu Gin Asn Ser Gin Val Phe Ser Leu He Arg Pro Asn Glu 305 310 315 320

Asn Pro Ala His Lys Ser Gin Leu Val Trp Met Ala Cys His Ser Ala 325 330 335

Ala Phe Glu Asp Leu Arg He Ser Ser Phe He Arg Gly Lys Lys Val 340 345 350

Val Pro Arg Gly Lys Leu Ser Thr Arg Gly Val Gin He Ala Ser Asn 355 360 365

Glu Asn Val Glu Ala Met Asp Ser Ser Thr Leu Lys Leu Arg Ser Arg 370 375 380

Tyr Trp Ala He Arg Thr Arg Ser Gly Gly Asn Thr Asn Gin Gin Lys 385 390 395 400

Ala Ser Ala Gly Gin He Ser Val Gin Pro Thr Phe Ser Val Gin Arg 405 410 415

Asn Leu Pro Phe Glu Arg Ala Thr Val Met Ala Ala Phe Ser Gly Asn 420 425 430

Asn Glu Gly Arg Thr Ser Asp Met Arg Thr Glu Val He Arg Met Met 435 440 445

Glu Ser Ala Lys Pro Glu Asp Leu Ser Phe Gin Gly Arg Gly Val Phe 450 455 460

Glu Leu Ser Asp Glu Lys Ala Thr Asn Pro He Val Pro Ser Phe Asp 465 470 475 480

Met Ser Asn Glu Gly Ser Tyr Phe Phe Gly Asp Asn Ala Glu Glu Tyr 485 490 495

Asp Asn He 500

Claims (76)

What is claimed is:

1. A recombinant swinepox virus comprising a foreign DNA sequence inserted into the swinepox virus genomic DNA, wherein the foreign DNA sequence is inserted within a Hindlll M fragment of the swinepox virus genomic DNA and is capable of being expressed in a swinepox virus infected host cell.

2. The recombinant swinepox virus of claim 1, wherein the foreign DNA sequence is inserted within an approximately 2 Kb Hindlll to Bglll subfragment of the

Hindlll M fragment of the swinepox virus genomic DNA.

3. The recombinant swinepox virus of claim 2, wherein the foreign DNA sequence is inserted within a Bglll site located within the approximately 2 Kb Hindlll to Bglll subfragment of the swinepox virus genomic DNA.

4. The recombinant swinepox virus of claim 1, wherein the foreign DNA sequence is inserted within a larger Hindlll to Bglll subfragment of the Hindlll M fragment of the swinepox virus genomic DNA.

5. The recombinant swinepox virus of claim 4, wherein the foreign DNA sequence is inserted within an AccI site located within the larger Hindlll to Bglll subfragment of the swinepox virus genomic DNA.

6. The recombinant swinepox virus of claim 1, further comprising a foreign DNA sequence inserted into an open reading frame encoding swinepox virus thymidine kinase.

7. The recombinant swinepox virus of claim 6, wherein the foreign DNA is inserted into a Ndel site located within the open reading frame encoding the swinepox virus thymidine kinase.

8. A recombinant swinepox virus comprising a foreign DΝA sequence inserted into the swinepox virus genomic DΝA, wherein the foreign DΝA sequence is inserted within a Hindlll Ν fragment of the swinepox virus genomic DΝA and is capable of being expressed in a swinepox virus infected host cell.

9. The recombinant swinepox virus of claim

8, wherein the foreign DΝA sequence is inserted within an approximately 2.0 kB Hindlll to BamHI subfragment of the

Hindlll Ν fragment of the swinepox virus genomic DΝA.

10. The recombinant swinepox virus of claim 8, wherein the foreign DΝA sequence is inserted within an approximately 1.2 kB BamHI to Hindlll subfragment of the Hindlll Ν fragment of the swinepox virus genomic DΝA.

11. The recombinant swinepox virus of claim

9, wherein the foreign DΝA sequence is inserted into an open reading frame within an approximately 2.0 kB Hindlll to BamHI subfragment of the Hindlll N fragment of the swinepox virus genomic DNA.

12. The recombinant swinepox virus of claim 11, wherein the open reading frame encodes a I7L gene.

13. The recombinant swinepox virus of claim

9, wherein the foreign DNA sequence is inserted within a EcoRV restriction endonuclease site within the approximately 2.0 kB Hindlll to BamHI subfragment of the swinepox virus genomic DNA.

14. The recombinant swinepox virus of claim 9, wherein the foreign DNA sequence is inserted within a SnaBI restriction endonuclease site within the approximately 2.0 kB Hindlll to BamHI subfragment of the swinepox virus genomic DNA.

15. The recombinant swinepox virus of claim

10, wherein the foreign DNA sequence is inserted into an open reading frame within an approximately 2.0 kB Hindlll to

BamHI subfragment of the Hindlll N fragment of the swinepox virus genomic DNA.

16. The recombinant swinepox virus of claim

15, wherein the open reading frame encodes a I4L gene.

17. The recombinant swinepox virus of claim 15, wherein the foreign DNA sequence is inserted within a Bglll restriction endonuclease site within the approximately 1.2 kB Hindlll to BamHI subfragment of the swinepox virus genomic DNA.

18. A recombinant swinepox virus comprising a foreign DNA sequence inserted into the swinepox virus genomic DNA, wherein the foreign DNA sequence is inserted within a Hindlll M fragment of the swinepox virus genomic DNA and is capable of being expressed in a swinepox virus infected host cell.

19. The recombinant swinepox virus of claim

18, wherein the foreign DNA sequence is inserted within an approximately 2.0 kB

Bglll to Hindu subfragment of the Hindlll M fragment of the swinepox virus genomic DNA.

20. The recombinant swinepox virus of claim

19, wherein the foreign DNA sequence is inserted within an approximately 3.6 kB larger Hindlll to Bglll subfragment of the Hindlll M fragment of the swinepox virus genomic DNA.

21. The recombinant swinepox virus of claim

20, wherein the foreign DNA sequence is inserted into an open reading frame within an approximately 2.0 kB Bglll to

Hindlll subfragment of the Hindlll M fragment of the swinepox virus genomic DNA.

22. The recombinant swinepox virus of claim 21, wherein the open reading frame encodes a I4L gene.

23. The recombinant swinepox virus of claim 21, wherein the open reading frame encodes a ol gene.

24. The recombinant swinepox virus of claim 19, wherein the foreign DNA sequence is inserted within a Bglll restriction endonuclease site within the approximately 2.0 kB Bglll to Hindlll subfragment of the swinepox virus genomic DNA.

25. The recombinant swinepox virus of claim

24, wherein the foreign DNA sequence is inserted into an open reading frame within the 3.6 kB larger Hindlll to Bglll subfragment of the Hindlll M fragment of the swinepox virus genomic DNA.

26. A recombinant swinepox virus comprising a foreign DNA sequence inserted into the swinepox virus genomic DNA, wherein the foreign DNA sequence is inserted within a

Hindlll K fragment of the swinepox virus genomic DNA and is capable of being expressed in a swinepox virus infected host cell.

27. The recombinant swinepox virus of claim 26, wherein the foreign DNA sequence is inserted into an approximately 3.2 kB subfragment of the Hindlll K fragment of the swinepox virus genomic DNA.

28. The recombinant swinepox virus of claim

27, wherein the foreign DNA sequence is inserted into an open reading frame within an approximately 3.2 kB subfragment of the Hindlll K fragment of the swinepox virus genomic DNA.

29. The recombinant swinepox virus of claim

28, wherein the open reading frame encodes a B18R gene.

30. The recombinant swinepox virus of claim 28, wherein the open reading frame encodes a B4R gene .

31. The recombinant swinepox virus of claim

1, wherein the foreign DNA sequence encodes a polypeptide.

32. The recombinant swinepox virus of claim 31, wherein the polypeptide is antigenic.

33. The recombinant fowlpox virus of claim 1, further comprising a foreign DNA sequence which encodes a detectable marker.

34. The recombinant fowlpox virus of claim 33, wherein the detectable marker is E. coli beta-galactosidase .

35. The recombinant fowlpox virus of claim

33, wherein the detectable marker is E. coli beta-glucuronidase.

36. The recombinant swinepox virus of claim

1, wherein the foreign DNA sequence encodes a cytokine.

37. The recombinant swinepox virus of claim

36, wherein the cytokine is chicken myelomonocytic growth factor (cMGF) or chicken interferon (cIFN) .

38. The recombinant swinepox virus of claim

36, wherein the cytokine is selected from a group consisting of interleukin-2, inter1eukin- 6 , inter1eukin- 12 , interferons, granulocyte-macrophage colony stimulating factors, and interleukin receptors.

39. The recombinant swinepox virus of claim

32, wherein the antigenic polypeptide is derived from the group consisting of: human herpesvirus, herpes simplex virus- 1, herpes simplex virus-2, human cytomegalovirus, Epstein-Barr virus, Varicell-Zoster virus, human herpesvirus- 6, human herpesvirus-7, human influenza, human immunodeficiency virus, rabies virus, measles virus, hepatitis B virus and hepatitis C virus.

40. The recombinant swinepox virus of claim

32, wherein the antigenic polypeptide is hepatitis B virus core protein or hepatitis B virus surface protein.

41. The recombinant swinepox virus of claim

32, wherein the antigenic polypeptide is equine influenza virus neuraminidase or hemagglutinin.

42. The recombinant swinepox virus of claim 32, wherein the antigenic polypeptide is selected from the group consisting of: equine influenza virus type A/Alaska 91 neuraminidase, equine influenza virus type A/Kentucky 92 neuraminidase, equine influenza virus type A/Prague 56 neuraminidase, equine influenza virus type A/Miami 63 neuraminidase, equine influenza virus type A/Kentucky 81 neuraminidase, equine herpesvirus type 1 glycoprotein B, and equine herpesvirus type 1 glycoprotein D.

43. The recombinant swinepox virus of claim 32, wherein the antigenic polypeptide is derived from the group consisting of: hog cholera virus gEl, hog cholera virus gE2, swine influenza virus hemagglutinin, neurominidase, matrix and nucleoprotein, pseudorabies virus gB, gC and gD, and PRRS virus 0RF7.

44. The recombinant swinepox virus of claim 32, wherein the antigenic polypeptide is selected from the group consisting of: Infectious bovine rhinotracheitis virus gE, bovine respiratory syncytial virus attachment protein (BRSV G) , bovine respiratory syncytial virus fusion protein (BRSV F) , bovine respiratory syncytial virus nucleocapsid protein

(BRSV N) , bovine parainfluenza virus type 3 fusion protein, and the bovine parainfluenza virus type 3 hemagglutinin neuraminidase.

45. The recombinant swinepox virus of claim 32, wherein the antigenic polypeptide is bovine viral diarrhea virus (BVDV) glycoprotein 48 or glycoprotein 53.

46. The recombinant swinepox virus of claim 32, wherein the foreign DNA sequence encodes an antigenic polypeptide which is derived or derivable from a group consisting of: feline immunodeficiency virus gag, feline immunodeficiency virus env, infectious laryngotracheitis virus glycoprotein B, infectious laryngotracheitis virus gl, infectious laryngotracheitis virus gD, infectious bovine rhinotracheitis virus glycoprotein G, infectious bovine rhinotracheitis virus glycoprotein E, pseudorabies virus glycoprotein 50, pseudorabies virus II glycoprotein B, pseudorabies virus III glycoprotein C, pseudorabies virus glycoprotein E, pseudorabies virus glycoprotein H, marek's disease virus glycoprotein A, marek's disease virus glycoprotein B, marek's disease virus glycoprotein D, newcastle disease virus hemagglutinin or neuraminadase, newcastle disease virus fusion, infectious bursal disease virus VP2 , infectious bursal disease virus VP3 , infectious bursal disease virus VP4, infectious bursal disease virus polyprotein, infectious bronchitis virus spike, infectious bronchitis virus matrix, and chick anemia virus.

47. The homology vector of claim 1, wherein the foreign DNA sequence is under control of a promoter.

48. The recombinant swinepox virus of claim 47, wherein the foreign DNA sequence is under control of an endogenous upstream poxvirus promoter.

49. The recombinant swinepox virus of claim 47, wherein the foreign DNA sequence is under control of a heterologous upstream promoter.

50. The recombinant swinepox virus of claim 47, wherein the promoter is selected from a group consisting of: synthetic pox viral promoter, pox synthetic late promoter 1, pox synthetic late promoter 2 early promoter 2, pox OIL promoter, pox I4L promoter, pox I3L promoter, pox I2L promoter, pox I1L promoter, and pox E10R promoter.

51. The recombinant swinepox virus of claim 1, which is designated S-SPV-042.

52. The recombinant swinepox virus of claim 1, which is designated S-SPV-043.

53. The recombinant swinepox virus of claim 1, which is designated S-SPV-041.

54. The recombinant swinepox virus of claim

1, which is designated S-SPV-045.

55. The recombinant swinpox virus of claim 1, designated S-SPV-046.

56. The recombinant swinpox virus of claim 1, designated S-SPV-047.

57. The recombinant swinpox virus of claim 1, designated S-SPV-048

58. The recombinant swinepox virus of claim 8, which is designated S-SPV-049.

59. The recombinant swinepox virus of claim 8, which is designated S-SPV-050.

60. The recombinant swinpox virus of claim 1, designated S-SPV-052.

61. The recombinant swinpox virus of claim 1, designated S-SPV-053.

62. The recombinant swinpox virus of claim 1, designated S-SPV-054.

63. The recombinant swinpox virus of claim 1, designated S-SPV-055.

64. The recombinant swinpox virus of claim 8, designated S-SPV-060.

65. The recombinant swinpox virus of claim 8, designated S-SPV-061.

66. The recombinant swinpox virus of claim 8, designated S-SPV-062.

67. A homology vector for producing a recombinant swinepox virus by inserting foreign DNA into the viral genome of a swinepox virus which comprises a double- stranded DNA molecule consisting essentially of:

a) double stranded foreign DNA not usually present within the swinepox virus viral genome;

b) at one end the foreign DNA, double- stranded swinepox virus DNA homologous to the viral genome located at one side of the Hindlll N fragment of the coding region of the swinepox virus viral genome,- and

c) at the other end of the foreign DNA, double-stranded swinepox virus DNA homologous to the viral genome located at the other side of the

Hindlll N fragment of the coding region of the swinepox virus viral genome.

68. The homology vector of claim 67, wherein the foreign DNA sequence encodes a cytokine.

69. The homology vector of claim 68, wherein the cytokine is chicken myelomonocytic growth factor (cMGF) or chicken interferon (cIFN) .

70. The homology vector of claim 67, wherein the foreign DNA sequence encodes a polypeptide.

71. A homology vector of claim 70, wherein the polypeptide is antigenic.

72. The homology vector of claim 67, wherein the foreign DNA sequence is under control of a promoter.

73. A vaccine useful for immunizing an animal against swinepox virus which comprises an effective immunizing amount of the recombinant swinepox virus of claims 1 and a suitable carrier.

74. A method of immunizing an animal against a human pathogen which comprises administering to the animal an effective immunizing dose of the vaccine of claim 73.

75. A method of immunizing an animal against an animal pathogen which comprises administering to the animal an effective immunizing dose of the vaccine of claim 73.

76. A method of enhancing an avian immune response which comprises administering to a person an effective dose of a recombinant swinepox virus of claim 1 and a suitable carrier.