Infectious cDNA of north american porcine reproductive and respiratory syndrome (PRRS) virus and uses thereof
View Patent ↗The invention provides isolated polynucleotide molecules that comprise a DNA sequence encoding an infectious RNA sequence encoding a genetically-modified North American PRRS virus, wherein the polynucleotide molecule lacks at least one detectable antigenic epitope of North American PRRS virus. The invention also provides vaccines comprising genetically modified North American PRRS virus, RNA molecules, plasmids and viral vectors comprising the isolated polynucleotide molecules. Also provided are isolated polynucleotide molecules further comprising at least one nucleotide sequence that encodes a detectable heterologous antigenic epitope, and vaccines comprising North American PRRS virus, RNA molecules, plasmids and viral vectors comprising such isolated polynucleotide molecules.
1. An isolated DNA molecule comprising a DNA sequence encoding an infectious RNA molecule encoding a Porcine Reproductive and Respiratory Syndrome (PRRS) virus, wherein said encoding DNA sequence is selected from the group consisting of:
(a) SEQ ID NO:1;
(b) a sequence that has at least 70% identity to SEQ ID NO:1; and
(c) a sequence that hybridizes to the complement of SEQ ID NO:1 under highly stringent conditions which comprise hybridization to filter bound DNA in 0.5M NaHPO 4 , 7% SDS, 1 mM EDTA at 65° C., and washing in 0.1×SSC/0.1% SDS at 68° C.; and
wherein, in all cases, said isolated DNA molecule (1) lacks an encoding DNA sequence corresponding to all of amino acids 616 through 752 (SEQ ID NO: 1 positions 3186-3596) or all of amino acids 628 through 759 (SEQ ID NO: 1 positions 3222-3617) in the hypervariable region of the nonstructural protein 2-coding region of PRRS ORF1a; (2) wherein said isolated DNA molecule does not comprises a nucleotide sequence, within said PRRS encoding sequence, that encodes a detectable heterologous antigenic epitope; and (3) wherein the DNA sequence than encodes the infectious RNA molecule is operably linked to a promoter.
2. The isolated polynucleotide molecule of claim 1 , wherein said molecule lacks the DNA sequence encoding for amino acids 616 through 752 in the hypervariable region of the nonstructural protein 2 coding region of ORF1a.
3. The isolated polynucleotide molecule of claim 1 , wherein said molecule lacks the DNA sequence encoding for amino acids 628 through 759 in the hypervariable region of the nonstructural protein 2 coding region of ORF1a.
4. An isolated polynucleotide molecule according to claim 1 , that is genetically modified such that when it infects a porcine animal it is unable to produce PRRS disease in the animal, yet is able to elicit an effective immunoprotective response against a PRRS virus in the animal.
5. An isolated polynucleotide molecule according to claim 1 that is at least 75% identical to SEQ ID NO: 1.
6. An isolated polynucleotide molecule according to claim 1 that is at least 85% identical to SEQ ID NO: 1.
7. An isolated PRRS virus encoded by the infectious RNA molecule claim 2 .
8. An isolated PRRS virus encoded by the infectious RNA molecule of claim 3 .