PMID- 24176053
OWN - NLM
STAT- MEDLINE
DCOM- 20140729
LR  - 20211021
IS  - 1297-9716 (Electronic)
IS  - 0928-4249 (Print)
IS  - 0928-4249 (Linking)
VI  - 44
IP  - 1
DP  - 2013 Oct 31
TI  - A novel porcine reproductive and respiratory syndrome virus vector system that 
      stably expresses enhanced green fluorescent protein as a separate transcription 
      unit.
PG  - 104
LID - 10.1186/1297-9716-44-104 [doi]
AB  - Here we report the rescue of a recombinant porcine reproductive and respiratory 
      syndrome virus (PRRSV) carrying an enhanced green fluorescent protein (EGFP) 
      reporter gene as a separate transcription unit. A copy of the transcription 
      regulatory sequence for ORF6 (TRS6) was inserted between the N protein and 3'-UTR 
      to drive the transcription of the EGFP gene and yield a general purpose 
      expression vector. Successful recovery of PRRSV was obtained using an RNA 
      polymerase II promoter to drive transcription of the full-length virus genome, 
      which was assembled in a bacterial artificial chromosome (BAC). The recombinant 
      virus showed growth replication characteristics similar to those of the wild-type 
      virus in the infected cells. In addition, the recombinant virus stably expressed 
      EGFP for at least 10 passages. EGFP expression was detected at approximately 10 h 
      post infection by live-cell imaging to follow the virus spread in real time and 
      the infection of neighbouring cells occurred predominantly through 
      cell-to-cell-contact. Finally, the recombinant virus generated was found to be an 
      excellent tool for neutralising antibodies and antiviral compound screening. The 
      newly established reverse genetics system for PRRSV could be a useful tool not 
      only to monitor virus spread and screen for neutralising antibodies and antiviral 
      compounds, but also for fundamental research on the biology of the virus.
FAU - Wang, Chengbao
AU  - Wang C
FAU - Huang, Baicheng
AU  - Huang B
FAU - Kong, Ning
AU  - Kong N
FAU - Li, Qiongyi
AU  - Li Q
FAU - Ma, Yuping
AU  - Ma Y
FAU - Li, Zhijun
AU  - Li Z
FAU - Gao, Jiming
AU  - Gao J
FAU - Zhang, Chong
AU  - Zhang C
FAU - Wang, Xiangpeng
AU  - Wang X
FAU - Liang, Chao
AU  - Liang C
FAU - Dang, Lu
AU  - Dang L
FAU - Xiao, Shuqi
AU  - Xiao S
FAU - Mu, Yang
AU  - Mu Y
FAU - Zhao, Qin
AU  - Zhao Q
FAU - Sun, Yani
AU  - Sun Y
FAU - Almazan, Fernando
AU  - Almazan F
FAU - Enjuanes, Luis
AU  - Enjuanes L
FAU - Zhou, En-Min
AU  - Zhou EM
AD  - Department of Preventive Veterinary Medicine, College of Veterinary Medicine, 
      Northwest A&F University, No, 22 Xinong Road, Yangling, Shaanxi 712100, China. 
      zhouem@nwsuaf.edu.cn.
LA  - eng
SI  - GENBANK/JX087437
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20131031
PL  - England
TA  - Vet Res
JT  - Veterinary research
JID - 9309551
RN  - 0 (Antibodies, Neutralizing)
RN  - 0 (Antibodies, Viral)
RN  - 0 (Antiviral Agents)
RN  - 0 (Genetic Markers)
RN  - 0 (Recombinant Proteins)
RN  - 0 (enhanced green fluorescent protein)
RN  - 147336-22-9 (Green Fluorescent Proteins)
SB  - IM
MH  - Animals
MH  - Antibodies, Neutralizing/*metabolism
MH  - Antibodies, Viral/*metabolism
MH  - Antiviral Agents/*pharmacology
MH  - Cell Line
MH  - Chromosomes, Artificial, Bacterial/genetics
MH  - *Gene Expression Regulation, Viral
MH  - Genetic Markers
MH  - Genetic Vectors/*genetics/metabolism
MH  - *Genome, Viral
MH  - Green Fluorescent Proteins/genetics/metabolism
MH  - Haplorhini
MH  - Molecular Sequence Data
MH  - Porcine respiratory and reproductive syndrome virus/drug 
      effects/genetics/*physiology
MH  - Recombinant Proteins/genetics/metabolism
MH  - Sequence Analysis, DNA/veterinary
MH  - Transfection/veterinary
MH  - *Virus Replication
PMC - PMC4176086
EDAT- 2013/11/02 06:00
MHDA- 2014/07/30 06:00
PMCR- 2013/01/01
CRDT- 2013/11/02 06:00
PHST- 2013/04/18 00:00 [received]
PHST- 2013/10/21 00:00 [accepted]
PHST- 2013/11/02 06:00 [entrez]
PHST- 2013/11/02 06:00 [pubmed]
PHST- 2014/07/30 06:00 [medline]
PHST- 2013/01/01 00:00 [pmc-release]
AID - 1297-9716-44-104 [pii]
AID - 10.1186/1297-9716-44-104 [doi]
PST - epublish
SO  - Vet Res. 2013 Oct 31;44(1):104. doi: 10.1186/1297-9716-44-104.