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1. WO1996004010 - VR-2332 VIRAL NUCLEOTIDE SEQUENCE AND METHODS OF USE

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[ EN ]
Claims:
1. A purified and isolated nucleic acid comprising a fragmentary portion of the VR-2332 genome between ORF 2 and ORF 7 and having a length sufficient to provide a nucleotide sequence that is unique with respect to the LV virus genome.

2. The nucleic acid as set forth in Claim 1 , said portion including a coding region for the expression of a polypeptide capable of inducing an anti-PRRS immune response in swine.

3. The nucleic acid as set forth in Claim 1 , including said portion selected from Sequence ID No. 1 and being sufficiently dissimilar from portions of Sequence ID. No. 14 to prevent PCR amplification of portions of said Sequence ID No. 1.

4. The nucleic acid as set forth in Claim 3, including said portion consisting essentially of a sequence selected from a group consisting of Sequence ID Nos. 2, 4, 6, 8, 10, 12, and combinations thereof, together with all complimentary strands and degenerate amino acid residue coding equivalencies that may be obtained by site-directed mutagenesis.

5. The nucleic acid as set forth in Claim 3, including said portion consisting essentially of a sequence selected from a group consisting of Sequence ID Nos. 2, 4, 6, 8, 10, and 12, as well as inverse complimentary sequences depending from Sequence ID Nos. 2, 4, 6, 8, 10, and 12.

6. The nucleic acid as set forth in Claim 5, said group consisting of Sequence ID No. 1 from positions 2783 to 2801 , the inverse compliment of Sequence ID No. 1 from positions 3271 to 3289, Sequence ID No. 1 from positions 2289 to 2307, the inverse compliment of Sequence ID No. 1 from positions 2862 to 2880, Sequence ID No. 14 from positions 14112 to

14131, the inverse compliment of Sequence ID No. 14 from positions 14551 to 14570, Sequence ID No, 14 from positions 14575 to 14594, and the inverse compliment of Sequence ID NO. 14 from positions 14955 to 14974, sequence ID No. 1 from positions 2814 to 2832, the inverse compliment of Sequence ID No. 1 from positions 3273 to 3291 , Sequence ID No. 1 from positions 2816 to

2834, and the inverse compliment of Sequence ID No. 1 from positions 3181 to 3198.

7. A chimeric vector for use in expressing viral proteins from a host cell, comprising a promoter and a termination sequence connected by a coding region insert including a fragmentary portion of the VR-2332 genome between ORF 2 and ORF 7, said insert having a length sufficient to provide a nucleotide sequence unique with respect to the LV virus genome, together with all degenerate amino acid residue coding equivalencies that may be obtained by site-directed mutagenesis.

8. The vector as set forth in Claim 7, including said insert consisting essentially of a sequence selected from a group consisting of Sequence ID Nos. 2, 4, 6, 8, 10, and 12, as well as inverse complimentary sequences depending from Sequence ID Nos. 2, 4, 6, 8, 10, and 12.

9. A vaccine for immunizing animals against a VR-2332 form of PRRS, comprising a polypeptide-coding region replicating a nucleotide sequence selected as a portion of Sequence ID No. 1 and having a length sufficient to provide a nucleotide sequence unique in comparison with respect to Sequence ID No. 14.

10. The vaccine as set forth in Claim 9, said coding region consisting essentially of a member selected from the group consisting of Sequence ID Nos. 2, 4, 6, 8, 10, and 12, and combinations thereof.

11. A vaccine for immunizing animals against a VR-2332 form of PRRS, comprising a VR-2332 amino acid residue sequence having a length sufficient to provide uniqueness in comparison with respect to LV virus amino acid residue sequences.

12. The vaccine as set forth in Claim 11 , said VR-2332 amino acid residue sequence consisting essentially of a sequence selected from the group consisting of Sequence ID Nos. 3, 5, 7, 9, 11, and 13, and combinations thereof.

13. A diagnostic assay for distinguishing between PRRS-causative viral strains, said assay comprising the steps of.
providing PCR oligonucleotide primers capable of selectively amplifying fragmentary genomic portions of a wild-type PRRS-causative virus;
obtaining a sample including cDNA derived from swine exhibiting PRRS clinical signs; and
using said primers in a polymerase chain reaction under conditions capable of selective amplification of cDNA from said PRRS- causative virus in said sample.

14. The assay as set forth in Claim 13, said PRRS-causative virus being selected from a group consisting of VR-2332 and LV virus.

15. The assay as set forth in Claim 14 including said primers being selected from a group consisting of fragmentary portions of Sequence ID No. 1, complimentary fragments of Sequence ID No. 1, fragmentary portions of Sequence ID No. 14, and complimentary fragments of Sequence ID No. 14, said primers being unique in comparison with respect to Sequence ID No. 14 when said primers derive from Sequence ID No. 1, said primers being unique in comparison with respect to Sequence ID No. 1 when said primers derive from Sequence ID No. 14;

16. A method of vaccinating an animal against VR-2332-caused PRRS, said method comprising the steps of:
providing a vaccine including at least one material selected from a group consisting of VR-2332 based polypeptides and VR-2332 based nucleic acids; and
administering said vaccine to said animal in a manner permitting said animal to develop an immune response to said material.