PMID- 19048364
OWN - NLM
STAT- MEDLINE
DCOM- 20090416
LR  - 20211020
IS  - 0920-8569 (Print)
IS  - 0920-8569 (Linking)
VI  - 38
IP  - 1
DP  - 2009 Feb
TI  - Insertion and deletion in a non-essential region of the nonstructural protein 2 
      (nsp2) of porcine reproductive and respiratory syndrome (PRRS) virus: effects on 
      virulence and immunogenicity.
PG  - 118-28
LID - 10.1007/s11262-008-0303-4 [doi]
AB  - Developing a vaccine that can differentiate infected and vaccinated animals 
      (DIVA) is a new challenge in the design of a vaccine for porcine reproductive and 
      respiratory syndrome virus (PRRSV). Nonstructural protein 2 (nsp2) is the single 
      largest viral product, and it has multiple roles in polypeptide processing and 
      replication complex formation. Using reverse genetics and an infectious PRRSV 
      cDNA clone, we constructed several deletion mutants in the non-essential region 
      of nsp2. One mutant, which has a 131 amino acid deletion within a relatively 
      conserved region of nsp2, was recovered and found to produce a viable virus. The 
      deleted region was replaced with a peptide tag encoding eight amino acids. A 
      recombinant virus containing the 131 amino acid deletion was found to produce 
      normal virus yields in MARC-145 cells and porcine alveolar macrophages (PAM); 
      however, gross and micro-histopathology showed that the virus was less virulent 
      in pigs. The 131 amino acid peptide was expressed as a recombinant protein and 
      used to coat enzyme-linked immunosorbent assay (ELISA) plates. This peptide was 
      recognized by sera from pigs infected with wild-type virus, but not by sera from 
      pigs infected with the deletion mutant. The results from this study show that 
      nsp2 is an important target for the development of marker vaccines and for virus 
      attenuation.
FAU - Kim, Dal-Young
AU  - Kim DY
AD  - Department of Diagnostic Medicine and Pathobiology, Kansas State University, 
      Manhattan, KS, 66506, USA. dykim@utmb.edu
FAU - Kaiser, Troy J
AU  - Kaiser TJ
FAU - Horlen, Kyle
AU  - Horlen K
FAU - Keith, Marcia L
AU  - Keith ML
FAU - Taylor, Lucas P
AU  - Taylor LP
FAU - Jolie, Rika
AU  - Jolie R
FAU - Calvert, Jay G
AU  - Calvert JG
FAU - Rowland, Raymond R R
AU  - Rowland RR
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, Non-P.H.S.
DEP - 20081202
PL  - United States
TA  - Virus Genes
JT  - Virus genes
JID - 8803967
RN  - 0 (Antibodies, Viral)
RN  - 0 (Vaccines, Marker)
RN  - 0 (Viral Nonstructural Proteins)
RN  - 0 (Virulence Factors)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Antibodies, Viral/blood
MH  - Cell Line
MH  - Cells, Cultured
MH  - Enzyme-Linked Immunosorbent Assay/methods
MH  - Lung/pathology
MH  - Macrophages, Alveolar/virology
MH  - Molecular Sequence Data
MH  - *Mutagenesis, Insertional
MH  - Porcine Reproductive and Respiratory Syndrome/pathology/virology
MH  - Porcine respiratory and reproductive syndrome 
      virus/genetics/*immunology/*pathogenicity
MH  - *Sequence Deletion
MH  - Swine
MH  - Vaccines, Marker/immunology
MH  - Viral Nonstructural Proteins/genetics/*immunology
MH  - Virulence
MH  - Virulence Factors/genetics/*physiology
EDAT- 2008/12/03 09:00
MHDA- 2009/04/17 09:00
CRDT- 2008/12/03 09:00
PHST- 2008/06/29 00:00 [received]
PHST- 2008/11/10 00:00 [accepted]
PHST- 2008/12/03 09:00 [pubmed]
PHST- 2009/04/17 09:00 [medline]
PHST- 2008/12/03 09:00 [entrez]
AID - 10.1007/s11262-008-0303-4 [doi]
PST - ppublish
SO  - Virus Genes. 2009 Feb;38(1):118-28. doi: 10.1007/s11262-008-0303-4. Epub 2008 Dec 
      2.