PMID- 17553585
OWN - NLM
STAT- MEDLINE
DCOM- 20071101
LR  - 20191210
IS  - 0168-1702 (Print)
IS  - 0168-1702 (Linking)
VI  - 128
IP  - 1-2
DP  - 2007 Sep
TI  - Expression and stability of foreign tags inserted into nsp2 of porcine 
      reproductive and respiratory syndrome virus (PRRSV).
PG  - 106-14
AB  - The nonstructural protein 2 (nsp2) of porcine reproductive and respiratory 
      syndrome virus (PRRSV) is the single largest protein produced during infection. 
      The cDNA of the pCMV-129 infectious PRRSV clone was modified for accepting 
      foreign tags by first creating unique Mlu I and SgrA I restrictions sites at 
      nucleotide (nt) positions 3219 and 3614, respectively, within the C-terminal 
      region of nsp2. cDNAs encoding oligo- and polypeptide tags, including FLAG, 
      enhanced green fluorescent protein (EGFP) and luciferase were inserted into the 
      newly created restriction sites. The results showed that only the EGFP-containing 
      genomes were properly expressed and produced virus. EGFP fluorescence, but not 
      EGFP immunoreactivity, was lost during passage of recombinant EGFP viruses in 
      culture. Sequencing of a fluorescent-negative EGFP virus showed that the EGFP 
      remained intact, except for the appearance of arginine to cysteine mutation at 
      position 96, which may interfere with chromophore formation or function.
FAU - Kim, Dal-Young
AU  - Kim DY
AD  - Department of Diagnostic Medicine and Pathobiology, Kansas State University, 
      Manhattan, KS 66506, USA.
FAU - Calvert, Jay G
AU  - Calvert JG
FAU - Chang, Kyeong-Ok
AU  - Chang KO
FAU - Horlen, Kyle
AU  - Horlen K
FAU - Kerrigan, Maureen
AU  - Kerrigan M
FAU - Rowland, Raymond R R
AU  - Rowland RR
LA  - eng
GR  - P20 RR17708/RR/NCRR NIH HHS/United States
GR  - P20-RR017686/RR/NCRR NIH HHS/United States
PT  - Evaluation Study
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, U.S. Gov't, Non-P.H.S.
DEP - 20070605
PL  - Netherlands
TA  - Virus Res
JT  - Virus research
JID - 8410979
RN  - 0 (Bacterial Proteins)
RN  - 0 (Oligopeptides)
RN  - 0 (Peptides)
RN  - 0 (enhanced green fluorescent protein)
RN  - 147336-22-9 (Green Fluorescent Proteins)
RN  - 98849-88-8 (FLAG peptide)
RN  - EC 1.13.12.- (Luciferases)
RN  - EC 3.1.21.- (endodeoxyribonuclease MluI)
RN  - EC 3.1.21.- (endodeoxyribonuclease SgrAI)
RN  - EC 3.1.21.4 (Deoxyribonucleases, Type II Site-Specific)
RN  - EC 3.4.22.- (Cysteine Endopeptidases)
RN  - EC 3.4.22.- (nsP2 proteinase)
SB  - IM
MH  - Animals
MH  - Bacterial Proteins/metabolism
MH  - Cell Line
MH  - Cysteine Endopeptidases/genetics/*metabolism
MH  - Deoxyribonucleases, Type II Site-Specific/metabolism
MH  - Gene Expression Regulation, Viral
MH  - Green Fluorescent Proteins/genetics/*metabolism
MH  - Luciferases/genetics/*metabolism
MH  - Microscopy, Confocal
MH  - Mutagenesis, Site-Directed
MH  - Oligopeptides
MH  - Peptides/genetics/*metabolism
MH  - Porcine respiratory and reproductive syndrome 
      virus/genetics/metabolism/*physiology
MH  - Recombination, Genetic
MH  - Reverse Transcriptase Polymerase Chain Reaction
MH  - Swine/virology
MH  - Virus Replication
EDAT- 2007/06/08 09:00
MHDA- 2007/11/02 09:00
CRDT- 2007/06/08 09:00
PHST- 2006/05/26 00:00 [received]
PHST- 2007/04/03 00:00 [revised]
PHST- 2007/04/16 00:00 [accepted]
PHST- 2007/06/08 09:00 [pubmed]
PHST- 2007/11/02 09:00 [medline]
PHST- 2007/06/08 09:00 [entrez]
AID - S0168-1702(07)00151-7 [pii]
AID - 10.1016/j.virusres.2007.04.019 [doi]
PST - ppublish
SO  - Virus Res. 2007 Sep;128(1-2):106-14. doi: 10.1016/j.virusres.2007.04.019. Epub 
      2007 Jun 5.