Infectious cDNA clone of North American porcine reproductive and respiratory syndrome (PRRS) virus and uses thereof

• US 6,500,662 B1
• Filed: 12/22/1999
• Issued: 12/31/2002
• Est. Priority Date: 12/22/1998
• Status: Expired due to Term

• Current Assignee: Pfizer Inc., Zoetis Services LLC (Zoetis Incorporated)
• Original Assignee: Pfizer Inc.
• Inventors: Welch, Siao-Kun W., Calvert, Jay G., Sheppard, Michael G.
• Primary Examiner(s): MOSHER, MARY
• Application Number:
  US09/470,661
• Time in Patent Office: 1,105 Days
• Field of Search: 536/23.72, 435/235.1, 435/325, 435/320.1, 424/204.1
• US Class Current: 435/235.1
• CPC Class Codes: A61K 2039/525 Virus
• CPC Class Codes: A61K 2039/5254 avirulent or attenuated
• CPC Class Codes: A61K 2039/5256 expressing foreign proteins
• CPC Class Codes: A61K 2039/53 DNA (RNA) vaccination
• CPC Class Codes: A61K 2039/552 Veterinary vaccine
• CPC Class Codes: A61K 39/12 Viral antigens
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 15/00 Drugs for genital or sexual disorders for disorders of sex hormones A61P5/24; Contraceptives
• CPC Class Codes: A61P 15/08 for gonadal disorders or for enhancing fertility, e.g. inducers of ovulation or of spermatogenesis
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 31/14 for RNA viruses
• CPC Class Codes: C07K 14/005 from viruses
• CPC Class Codes: C12N 15/86 Viral vectors
• CPC Class Codes: C12N 2770/10021 Viruses as such, e.g. new isolates, mutants or their genomic sequences
• CPC Class Codes: C12N 2770/10034 Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
• CPC Class Codes: C12N 2770/10043 viral genome or elements thereof as genetic vector
• CPC Class Codes: C12N 2770/10061 Methods of inactivation or attenuation
• CPC Class Codes: C12N 2770/10062 by genetic engineering
• CPC Class Codes: C12N 7/00 Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00

Abstract

The invention provides isolated polynucleotide molecules, including plasmids; viral vectors; and transfected host cells that comprise a DNA sequence encoding an infectious RNA sequence encoding a North American PRRS virus; and also North American PRRS viruses encoded thereby. The invention further provides isolated infectious RNA molecules encoding a North American PRRS virus. The invention also provides isolated polynucleotide molecules, infectious RNA molecules, viral vectors, and transfected host cells encoding genetically-modified North American PRRS viruses; and genetically-modified North American PRRS viruses encoded thereby. The invention also provides vaccines comprising such plasmids, RNA molecules, viral vectors, and North American PRRS viruses, and methods of using these vaccines in swine and in other animals. Also provided are isolated polynucleotide molecules, viral vectors, and transfected host cells that comprise a nucleotide sequence encoding a peptide of a North American PRRS virus. These viral vectors and transfected host cell lines are useful in providing peptides to compensate for mutated peptide coding sequences of DNA sequences encoding genetically-modified North American PRRS viruses so that functional virions can be generated.

Claims

1. An isolated polynucleotide molecule comprising a DNA sequence encoding an infectious RNA molecule encoding a North American PRRS virus, wherein said DNA sequence is SEQ ID NO:

1 or a sequence that hybridizes to the complement of SEQ ID NO;

1 under highly stringent condition, wherein said highly stringent conditions comprise hybridization to filter-bound DNA in 0.5 M NaHPO₄, 7% SDS, 1 mM EDTA at 65°

C., and washing in 0.1×

SSC/0.1% SDS at 68°

C.

2. A transfected host cell comprising a DNA sequence encoding an infectious RNA molecule encoding a North American PRRS virus, wherein said DNA sequence is SEQ ID NO:

1 or a sequence that hybridizes to the complement of SEQ ID NO;

1 under highly stringent condition, wherein said highly stringent conditions comprise hybridization to filter-bound DNA in 0.5 M NaHPO₄, 7% SDS, 1 mM EDTA at 65°

C., and washing in 0.1×

SSC/0.1% SDS at 68°

C., which transfected host cell is capable of expressing the encoded North American PRRS virus.

3. An isolated polynucleotide molecule comprising a DNA sequence encoding an infectious RNA molecule encoding a North American PRRS virus, wherein said DNA sequence is SEQ ID NO:

1.

4. An isolated polynucleotide molecule in the form of a plasmid, wherein said isolated polynucleotide molecule comprises a DNA sequence encoding an infectious RNA molecule encoding a North American PRRS virus, wherein said DNA sequence is SEQ ID NO:

1.

5. An isolated infectious RNA molecule encoded by an isolated polynucleotide molecule comprising SEQ ID NO:

1, which infectious RNA molecule encodes a North American PRRS virus.

6. A recombinant North American PRRS virus encoded by an isolated polynucleotide molecule comprising a DNA sequence encoding an infectious RNA molecule encoding a North American PRRS virus, wherein said DNA sequence is SEQ ID NO:

1.