North American Porcine Reproductive and Respiratory Syndrome (PRRS) Virus and Uses Thereof
AssigneeZoetis Services LLC
InventorsJay Gregory CALVERT, David Ewell SLADE, Siao-Kun W. WELCH
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Abstract
The invention provides isolated polynucleotide molecules that comprise a DNA sequence encoding an infectious RNA sequence encoding a genetically-modified North American PRRS virus, methods to make it and related polypeptides, polynucleotides, and various components. Vaccines comprising the genetically modified virus and polynucleotides and a diagnostic kit to distinguish between naturally infected and vaccinated animals are also provided.
Claims8
An isolated Porcine Reproductive and Respiratory Syndrome Virus (PRRS) wherein the protein thereof encoded by ORF1a is selected from a group consisting of those amino acid sequences that contain any of:
- an amino acid N within the amino acid sequence AMANVYD (SEQ ID NO:
- 9);
- an amino acid N within the amino acid sequence IGHNAVM (SEQ ID NO:
- 12);
- an amino acid D within the amino acid sequence TVPDGNC (SEQ ID NO:
- 15);
- an amino acid Y within the amino acid sequence CWWYLFD (SEQ ID NO:
- 18);
- an amino acid H within the amino acid sequence HGVHGKY (SEQ ID NO:
- 21);
- an amino acid V within the amino acid sequence AAKVDQY (SEQ ID NO:
- 24);
- an amino acid T within the amino acid sequence PSATDTS (SEQ ID NO:
- 27);
- an amino acid L within the amino acid sequence LNSLLSK (SEQ ID NO:
- 30).
- an amino acid C within the amino acid sequence APMCQDE (SEQ ID NO:
- 33);
- an amino acid T within the amino acid sequence CAPTGMD (SEQ ID NO:
- 36);
- an amino acid A within the amino acid sequence PKVAKVS (SEQ ID NO:
- 39);
- an amino acid I within the amino acid sequence AGEIVGV (SEQ ID NO:
- 42);
- an amino acid N within the amino acid sequence ADFNPEK (SEQ ID NO:
- 45);
- and
- an amino acid I within the amino acid sequence QTPILGR (SEQ ID NO:
- 48).
A plasmid capable of directly transfecting a suitable host cell and expressing a Porcine Reproductive and Respiratory Syndrome Virus (PRRS) from the suitable host cell so transfected, which plasmid comprises:
- (a) a DNA sequence encoding an infectious RNA molecule encoding the PRRS virus, and (b) a promoter capable of transcribing said infectious RNA molecule, wherein the protein encoded by ORF1a of said virus has an amino acid sequence that contains:
- (1) an amino acid N within the amino acid sequence ANV (see SEQ ID NO:
- 9);
- an amino acid N within the amino acid sequence HNA (see SEQ ID NO:
- 12);
- an amino acid D within the amino acid sequence PDG (see SEQ ID NO:
- 15),
- an amino acid Y within the amino acid sequence WYL (see SEQ ID NO:
- 18);
- an amino acid H within the amino acid sequence VHG (see SEQ ID NO:
- 21), or any subset thereof;
- and/or
- (2) an amino acid V within the amino acid sequence KVD (see SEQ ID NO:
- 24);
- an amino acid T within the amino acid sequence ATD (see SEQ ID NO:
- 27);
- an amino acid L within the amino acid sequence SLL (see SEQ ID NO:
- 30).
- an amino acid C within the amino acid sequence MCQ (see SEQ ID NO:
- 33), or any subset thereof;
- and/or
- (3) an amino acid T within the amino acid sequence PTG (see SEQ ID NO:
- 36);
- an amino acid A within the amino acid sequence VAK (see SEQ ID NO:
- 39), or any subset thereof;
- and/or
- (4) an amino acid I within the amino acid sequence EIV (see SEQ ID NO:
- 42);
- an amino acid N within the amino acid sequence FNP (see SEQ ID NO:
- 45);
- and amino acid I within the amino acid sequence PIL (see SEQ ID NO:
- 48), or any subset thereof.
An isolated polynucleotide molecule comprising a DNA sequence encoding an infectious RNA molecule encoding a North American PRRS virus, wherein said DNA sequence is selected from the group consisting of:
- (a) SEQ ID NO:6;
- (b) a sequence that has at least 85% identity to the DNA sequence of (a) wherein the protein encoded by ORF1a thereof has an amino acid sequence that contains:
- (1) an amino acid N within the amino acid sequence ANV (see SEQ ID NO:
- 9);
- an amino acid N within the amino acid sequence HNA (see SEQ ID NO:
- 12);
- an amino acid D within the amino acid sequence PDG (see SEQ ID NO:
- 15),
- an amino acid Y within the amino acid sequence WYL (see SEQ ID NO:
- 18);
- an amino acid H within the amino acid sequence VHG (see SEQ ID NO:
- 21), or any subset thereof;
- or
- (2) an amino acid V within the amino acid sequence KVD (see SEQ ID NO:
- 24);
- an amino acid T within the amino acid sequence ATD (see SEQ ID NO:
- 27);
- an amino acid L within the amino acid sequence SLL (see SEQ ID NO:
- 30).
- an amino acid C within the amino acid sequence MCQ (see SEQ ID NO:
- 33), or any subset thereof;
- or
- (3) an amino acid T within the amino acid sequence PTG (see SEQ ID NO:
- 36);
- an amino acid A within the amino acid sequence VAK (see SEQ ID NO:
- 39), or any subset thereof;
- or
- (4) an amino acid I within the amino acid sequence EIV (see SEQ ID NO:
- 42);
- an amino acid N within the amino acid sequence FNP (see SEQ ID NO:
- 45);
- and amino acid I within the amino acid sequence PIL (see SEQ ID NO:
- 48), or any subset thereof;
- and
- (c) a DNA sequence that hybridizes to the complement of a DNA sequence of (a) or (b) under highly stringent conditions which comprise hybridization to filter bound DNA in 0.5 M NaHPo4, 7% SDS, 1 mM EDTA at 65 degrees C., and washing in 0.1 SSC/0/1% SDS at 68 degrees C.
Dependencies are inferred from the claim text.