PMID- 16378975
OWN - NLM
STAT- MEDLINE
DCOM- 20060127
LR  - 20181113
IS  - 0022-538X (Print)
IS  - 1098-5514 (Electronic)
IS  - 0022-538X (Linking)
VI  - 80
IP  - 2
DP  - 2006 Jan
TI  - Identification of 5' and 3' cis-acting elements of the porcine reproductive and 
      respiratory syndrome virus: acquisition of novel 5' AU-rich sequences restored 
      replication of a 5'-proximal 7-nucleotide deletion mutant.
PG  - 723-36
AB  - We here demonstrate the successful engineering of the RNA genome of porcine 
      reproductive and respiratory syndrome virus (PRRSV) by using an infectious cDNA 
      as a bacterial artificial chromosome. Runoff transcription from this cDNA by SP6 
      polymerase resulted in capped synthetic RNAs bearing authentic 5' and 3' ends of 
      the viral genome that had specific infectivities of >5 x 10(5) PFU/microg of RNA. 
      The synthetic viruses recovered from the transfected cells were genotypically and 
      phenotypically indistinguishable from the parental virus. Using our system, a 
      series of genomic RNAs with nucleotide deletions in their 5' ends produced 
      viruses with decreased or no infectivity. Various pseudorevertants were isolated, 
      and acquisition of novel 5' sequences of various sizes, composed predominantly of 
      A and U bases, restored their infectivities, providing a novel insight into 
      functional elements of the 5' end of the PRRSV genome. In addition, our system 
      was further engineered to generate a panel of self-replicating, self-limiting, 
      luciferase-expressing PRRSV viral replicons bearing various deletions. Analysis 
      of these replicons revealed the presence and location of a 3' cis-acting element 
      in the genome that was required for replication. Moreover, we produced enhanced 
      green fluorescent protein-expressing infectious viruses, which indicates that the 
      PRRSV cDNA/viral replicon/recombinant virus can be developed as a vector for the 
      expression of a variety of heterologous genes. Thus, our PRRSV reverse genetics 
      system not only offers a means of directly investigating the molecular mechanisms 
      of PRRSV replication and pathogenesis but also can be used to generate new 
      heterologous gene expression vectors and genetically defined antiviral vaccines.
FAU - Choi, Yu-Jeong
AU  - Choi YJ
AD  - Department of Microbiology, College of Medicine and Medical Research Institute, 
      Chungbuk National University, 12 Gaeshin-Dong, Heungduk-Ku, Cheongju, Korea.
FAU - Yun, Sang-Im
AU  - Yun SI
FAU - Kang, Shien-Young
AU  - Kang SY
FAU - Lee, Young-Min
AU  - Lee YM
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - J Virol
JT  - Journal of virology
JID - 0113724
RN  - 0 (DNA, Complementary)
RN  - 0 (RNA, Viral)
RN  - 0 (enhanced green fluorescent protein)
RN  - 147336-22-9 (Green Fluorescent Proteins)
RN  - EC 1.13.12.- (Luciferases)
SB  - IM
MH  - 3' Flanking Region/*genetics
MH  - 5' Flanking Region/*genetics
MH  - Animals
MH  - Cell Line
MH  - DNA, Complementary/biosynthesis
MH  - Genetic Vectors
MH  - Green Fluorescent Proteins/metabolism
MH  - Luciferases/metabolism
MH  - Mutation
MH  - Porcine respiratory and reproductive syndrome virus/*genetics/growth & 
      development/metabolism
MH  - RNA, Viral/*biosynthesis/genetics
MH  - Replicon
PMC - PMC1346850
EDAT- 2005/12/28 09:00
MHDA- 2006/01/28 09:00
PMCR- 2006/05/01
CRDT- 2005/12/28 09:00
PHST- 2005/12/28 09:00 [pubmed]
PHST- 2006/01/28 09:00 [medline]
PHST- 2005/12/28 09:00 [entrez]
PHST- 2006/05/01 00:00 [pmc-release]
AID - 80/2/723 [pii]
AID - 1478-04 [pii]
AID - 10.1128/JVI.80.2.723-736.2006 [doi]
PST - ppublish
SO  - J Virol. 2006 Jan;80(2):723-36. doi: 10.1128/JVI.80.2.723-736.2006.