PMID- 22513707
OWN - NLM
STAT- MEDLINE
DCOM- 20150407
LR  - 20191210
IS  - 1873-2542 (Electronic)
IS  - 0378-1135 (Linking)
VI  - 159
IP  - 1-2
DP  - 2012 Sep 14
TI  - Stable expression of foreign gene in nonessential region of nonstructural protein 
      2 (nsp2) of porcine reproductive and respiratory syndrome virus: applications for 
      marker vaccine design.
PG  - 1-10
LID - 10.1016/j.vetmic.2012.03.015 [doi]
AB  - The nonstructural protein 2 (nsp2) of porcine reproductive and respiratory 
      syndrome virus (PRRSV) has been shown to be highly heterogeneous and variable 
      among PRRSV strains and some sequences in the middle region of the nsp2 are not 
      essential to viral replication. Recent studies have attempted to insert foreign 
      genes in the nsp2 nonessential regions but the foreign genes were not stably 
      expressed by recombinant viruses in vitro. In the present study, we first 
      constructed an infectious cDNA clone with deletion of 75 nucleotides (25 amino 
      acids) in the nsp2 region (rHuN4-F112-Delta508-532) of the attenuated vaccine virus 
      HuN4-F112 derived from a highly pathogenic PRRSV HuN4 and then inserted a gene 
      fragment encoding a immunodominant B-cell epitope (49 amino acids) of Newcastle 
      disease virus (NDV) nucleoprotein (NP) in-frame into the deletion site. The 
      viable recombinant virus was rescued from the full-length cDNA infectious clone 
      in vitro. The engineered viruses rescued from the cDNA clone indicated that the 
      deletions of 75 nucleotides and insertion of NDV NP gene in the nsp2 region did 
      not affect viral replication; they had similar growth kinetics to its parental 
      virus. The inserting gene could be expressed consistently when the recombinant 
      virus was passaged up to twenty times in cell cultures as determined by 
      immunofluorescence assay (IFA) and genomic sequencing. To investigate the 
      potential application of the NDV NP gene-inserted PRRSV as a marker vaccine, 
      piglets were immunized with the recombinant virus and then challenged with lethal 
      dose of highly pathogenic PRRSV. The immunized piglets produced specific 
      antibodies against both the NDV NP and PRRSV, and lacked antibodies against the 
      deleted 25aa nsp2 epitope. After challenge, all immunized piglets were protected 
      from clinical disease or death, while all piglets in control group died (5/5) by 
      ten days post challenge. The results of the present study indicated that the 
      recombinant PRRSV (rHuN4-F112-Delta508-532) could be used as a potential marker 
      vaccine against PRRS.
CI  - Copyright (c) 2012 Elsevier B.V. All rights reserved.
FAU - Xu, Yan-Zhao
AU  - Xu YZ
AD  - Division of Swine Infectious Diseases, Shanghai Veterinary Research Institute, 
      CAAS, Shanghai 200241, China.
FAU - Zhou, Yan-Jun
AU  - Zhou YJ
FAU - Zhang, Shan-Rui
AU  - Zhang SR
FAU - Jiang, Yi-Feng
AU  - Jiang YF
FAU - Tong, Wu
AU  - Tong W
FAU - Yu, Hai
AU  - Yu H
FAU - Tong, Guang-Zhi
AU  - Tong GZ
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20120330
PL  - Netherlands
TA  - Vet Microbiol
JT  - Veterinary microbiology
JID - 7705469
RN  - 0 (Antibodies, Viral)
RN  - 0 (Recombinant Proteins)
RN  - 0 (Vaccines, Marker)
RN  - 0 (Viral Nonstructural Proteins)
RN  - 0 (Viral Vaccines)
SB  - IM
MH  - Animals
MH  - Antibodies, Viral/blood
MH  - Body Temperature
MH  - Cell Line
MH  - Chlorocebus aethiops
MH  - Cricetinae
MH  - *Gene Expression
MH  - Mutagenesis, Insertional
MH  - Porcine Reproductive and Respiratory Syndrome/immunology
MH  - Porcine respiratory and reproductive syndrome virus/*genetics/*immunology
MH  - Random Allocation
MH  - Recombinant Proteins/genetics
MH  - Swine/immunology
MH  - Vaccines, Marker/genetics/immunology
MH  - Viral Nonstructural Proteins/*genetics
MH  - Viral Vaccines/*genetics/*immunology
EDAT- 2012/04/20 06:00
MHDA- 2015/04/08 06:00
CRDT- 2012/04/20 06:00
PHST- 2011/12/06 00:00 [received]
PHST- 2012/03/03 00:00 [revised]
PHST- 2012/03/09 00:00 [accepted]
PHST- 2012/04/20 06:00 [entrez]
PHST- 2012/04/20 06:00 [pubmed]
PHST- 2015/04/08 06:00 [medline]
AID - S0378-1135(12)00182-4 [pii]
AID - 10.1016/j.vetmic.2012.03.015 [doi]
PST - ppublish
SO  - Vet Microbiol. 2012 Sep 14;159(1-2):1-10. doi: 10.1016/j.vetmic.2012.03.015. Epub 
      2012 Mar 30.