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Proteins encoded by polynucleic acids of porcine reproductive and respiratory syndrome virus (PRRSV)

a technology of polynucleic acids, which is applied in the field of polynucleic acids isolated from porcine reproductive and respiratory syndrome viruses, can solve the problems of respiratory disease now becoming a serious problem, prrsv causes significant economic losses from pneumonia in nursery pigs, and the information regarding the sg mrnas of prrsv strains is very limited

Inactive Publication Date: 2006-03-23
PAUL PREM +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The vaccine effectively protects pigs against PRRSV infection by inducing antibodies and reducing the severity of clinical symptoms, and the DNA sequence aids in distinguishing the ISU-55 strain, addressing the molecular characterization and vaccine needs.

Problems solved by technology

However, information regarding the sg mRNAs of PRRSV strains, especially the U.S. PRRSV, is very limited.
PRRSV causes significant economic losses from pneumonia in nursery pigs (the exact extent of which are not fully known).
Respiratory disease has now become the main problem associated with PRRSV, due to the increasing prevalence of relatively high virulence strains of PRRSV.

Method used

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  • Proteins encoded by polynucleic acids of porcine reproductive and respiratory syndrome virus (PRRSV)
  • Proteins encoded by polynucleic acids of porcine reproductive and respiratory syndrome virus (PRRSV)
  • Proteins encoded by polynucleic acids of porcine reproductive and respiratory syndrome virus (PRRSV)

Examples

Experimental program
Comparison scheme
Effect test

example 1

Summary:

[0155] The sequences of ORFs 2 to 5 of one low virulence, one “moderate” virulence and one high virulence U.S. PRRSV isolate have been determined and analyzed. Comparisons with known sequences of other PRRSV isolates show that considerable sequence variations at both nucleotide and amino acid levels exist in ORFs 2 to 5 of seven U.S. isolates with differing virulence. However, ORFs 6 and 7 of these seven U.S. isolates are highly conserved (U.S. application Ser. No. 08 / 301,435). Extensive sequence variations were also found in ORFs 2 to 7 between the European LV and the U.S. isolates. The least virulent U.S. PRRSV isolate known (ISU-3927) displayed the most sequence variation, in comparison with other U.S. isolates.

[0156] The phylogenetic relationship of the U.S. isolates was also analyzed. Phylogenetic analysis of the ORFs 2 to 7 of the U.S. isolates indicated that there are at least three groups of PRRSV variants (or minor genotypes) within the major U.S. PRRSV genotype....

example 2

[0194] During the replication of PRRSV, six subgenomic mRNAs (sg mRNAs), in addition to the genomic RNA, are synthesized. These sg mRNAs were characterized in this experiment.

[0195] The sg mRNAs of PRRSV form a 3′-coterminal nested set in PRRSV-infected cells. Each of these sg mRNAs is polycistronic and contains multiple open reading frames, except for sg mRNA 7 (as shown by Northern blot analysis using ORF-specific probes). The sg mRNAs were not packaged into virions, and only the genomic RNA was detected in purified virions, suggesting that the encapsidation signal of PRRSV is likely localized in the ORF 1 region.

[0196] The numbers of sg mRNAs in PRRSV-infected cells varies among PRRSV isolates with differing virulence. An additional species of sg mRNA in some PRRSV isolates was shown in Experiment 1 above to be derived from the sequence upstream of ORF 4, and has been designated as sg mRNA 3-1.

[0197] The leader-mRNA junction sequences of sg mRNAs 3 and 4 of isolates ISU 79 and...

example 3

[0241] Cell line ATCC CRL 11171 was used for the propagation of PRRSV isolates. The maintenance of the cell line and isolation of virus were the same as previously described (Meng et al., J. Gen. Virol. 75:1795-1801 (1994); Meng et al., J. of Veterinary Diagnostic Investigation 8:374-381 (1996). Plasmacytoma cell line SP2 / O was used for cell fusion in MAb preparation. PRRSV ATCC VR 2385 was used as antigen for screening of hybridomas secreting PRRSV specific monoclonal antibodies.

[0242] Indirect Immunofluorescence Assay (IFA). Monolayers of ATCC CRL 11171 cells were inoculated with PRRSV VR 2385 at 0.1 multiplicities of infection, incubated for 48 hrs and fixed with methanol. Hybridoma supernatant was incubated on the fixed-cell monolayer at 37° C. for 30 min. Fluorescein-labeled goat anti-mouse IgG (H+L) conjugate was used to detect the specific reaction. One PRRSV N(ORF 7 products) specific monoclonal antibody, PP7eF11 was used as a positive control and cell culture supernatant f...

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Abstract

The present invention provides an isolated DNA sequence encoding, for example, at least one polypeptide selected from the group consisting of proteins encoded by one or more open reading frames (ORF's) of an Iowa strain of porcine reproductive and respiratory syndrome virus (PRRSV), specifically ISU-12, and the polypeptides encoded by the isolated DNA sequences. The present invention also concerns a vaccine comprising an effective amount of such a protein; methods of producing antibodies which specifically bind to such a protein; and methods of protecting a pig against a PRRSV, and treating a pig infected by a PRRSV.

Description

BACKGROUND OF THE INVENTION [0001] 1. Field of the Invention [0002] The present invention concerns polynucleic acids isolated from a porcine reproductive and respiratory syndrome virus (PRRSV), a protein and / or a polypeptide encoded by the polynucleic acids, a vaccine which protects pigs from a PRRSV based on the protein or polynucleic acids, methods of making the proteins, polypeptides and polynucleic acids, a method of protecting a pig from PRRS using the vaccine, a method of producing the vaccine, a method of treating a pig infected by or exposed to a PRRSV, and a method of detecting a PRRSV. [0003] 2. Discussion of the Background [0004] Porcine reproductive and respiratory syndrome (PRRS), a new and severe disease in swine, was first reported in the U.S.A. in 1987, and was rapidly recognized in many western European countries (reviewed by Goyal, J. Vet. Diagn. Invest., 1993, 5:656-664; and in U.S. application Ser. Nos. 08 / 131,625 and 08 / 301,435). The disease is characterized by ...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61K39/12C07H21/04C12N15/86C07K14/17A61K38/00G01N33/53A61K39/00A61P31/04A61P31/12C07K7/08C07K14/00C07K14/01C07K14/08C07K16/00C12N7/00C12N7/01C12N15/09C12N15/40C12P21/02C12P21/08G01N33/569
CPCA61K38/00A61K39/00A61K39/12A61K2039/51A61K2039/522A61K2039/5254G01N33/56983C12N7/00C12N2770/10021C12N2770/10022C12N2770/10034C12N2770/10064C07K14/005A61K2039/552A61P31/04A61P31/12
Inventor PAUL, PREMZHANG, YANJIN
Owner PAUL PREM
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