PMID- 11602771
OWN - NLM
STAT- MEDLINE
DCOM- 20011204
LR  - 20200217
IS  - 0022-1317 (Print)
IS  - 0022-1317 (Linking)
VI  - 82
IP  - Pt 11
DP  - 2001 Nov
TI  - Viable porcine arteriviruses with deletions proximal to the 3' end of the genome.
PG  - 2607-2614
LID - 10.1099/0022-1317-82-11-2607 [doi]
AB  - In order to obtain attenuated live vaccine candidates of porcine reproductive and 
      respiratory syndrome virus (PRRSV), a series of deletions was introduced at the 
      3' end of the viral genome using an infectious cDNA clone of the Lelystad virus 
      isolate. RNA transcripts from the full-length cDNA clones were transfected into 
      BHK-21 cells. The culture supernatant of these cells was subsequently used to 
      infect porcine alveolar macrophages to detect the production of progeny virus. It 
      is shown that C-terminal truncation of the nucleocapsid (N) protein, encoded by 
      ORF7, was tolerated for up to six amino acids without blocking the production of 
      infectious virus. Mutants containing larger deletions produced neither virus nor 
      virus-like particles containing viral RNA. Deletion analysis of the 3' UTR 
      immediately downstream of ORF7 showed that infectious virus was still produced 
      after removal of seven nucleotides behind the stop codon of ORF7. Deletion of 32 
      nucleotides in this region abolished RNA replication and, consequently, no 
      infectious virus was formed. Serial passage on porcine alveolar macrophages 
      demonstrated that the viable deletion mutants were genetically stable at the site 
      of mutation. In addition, the deletions did not affect the growth properties of 
      the recombinant viruses in vitro, while their antigenic profiles were similar to 
      that of wild-type virus. Immunoprecipitation experiments with the six-residue N 
      protein-deletion mutant confirmed that the truncated protein was indeed smaller 
      than the wild-type N protein. The deletion mutants produced in this study are 
      interesting candidate vaccines to prevent PRRS disease in pigs.
FAU - Verheije, M H
AU  - Verheije MH
AD  - Department of Infectious Diseases and Food Chain Quality, Institute for Animal 
      Science and Health, Lelystad, The Netherlands1.
FAU - Kroese, M V
AU  - Kroese MV
AD  - Department of Infectious Diseases and Food Chain Quality, Institute for Animal 
      Science and Health, Lelystad, The Netherlands1.
FAU - Rottier, P J M
AU  - Rottier PJM
AD  - Virology Division, Department of Infectious Diseases and Immunology, Veterinary 
      Faculty, Utrecht University, Utrecht, The Netherlands2.
FAU - Meulenberg, J J M
AU  - Meulenberg JJM
AD  - Department of Infectious Diseases and Food Chain Quality, Institute for Animal 
      Science and Health, Lelystad, The Netherlands1.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - J Gen Virol
JT  - The Journal of general virology
JID - 0077340
RN  - 0 (3' Untranslated Regions)
RN  - 0 (DNA, Complementary)
RN  - 0 (Nucleocapsid Proteins)
SB  - IM
MH  - 3' Untranslated Regions
MH  - Animals
MH  - Base Sequence
MH  - Cells, Cultured
MH  - Cloning, Molecular
MH  - DNA, Complementary/genetics
MH  - *Genome, Viral
MH  - Macrophages, Alveolar/virology
MH  - Molecular Sequence Data
MH  - Nucleocapsid Proteins/chemistry/genetics
MH  - Open Reading Frames
MH  - Porcine Reproductive and Respiratory Syndrome/virology
MH  - Porcine respiratory and reproductive syndrome virus/*genetics/*growth & 
      development/pathogenicity
MH  - Sequence Analysis, DNA
MH  - *Sequence Deletion
MH  - Swine
MH  - Transfection
EDAT- 2001/10/17 10:00
MHDA- 2002/01/05 10:01
CRDT- 2001/10/17 10:00
PHST- 2001/10/17 10:00 [pubmed]
PHST- 2002/01/05 10:01 [medline]
PHST- 2001/10/17 10:00 [entrez]
AID - 10.1099/0022-1317-82-11-2607 [doi]
PST - ppublish
SO  - J Gen Virol. 2001 Nov;82(Pt 11):2607-2614. doi: 10.1099/0022-1317-82-11-2607.