PMID- 23957156
OWN - NLM
STAT- MEDLINE
DCOM- 20131108
LR  - 20171116
IS  - 0001-6209 (Print)
IS  - 0001-6209 (Linking)
VI  - 53
IP  - 5
DP  - 2013 May 4
TI  - [Establishment of a porcine CD151 transgenic PK-15 cell line susceptible to 
      porcine reproductive and respiratory syndrome virus].
PG  - 507-14
AB  - OBJECTIVE: In order to study the role of porcine CD151 in infection of porcine 
      cells by porcine reproductive and respiratory syndrome virus (PRRSV), we 
      established a porcine CD151 transgenic PK-15 cell line. METHODS: The full-length 
      complementary DNA (cDNA) for porcine CD151 was amplified from porcine alveolar 
      macrophages by reverse transcription-polymerase chain reaction (RT-PCR), and 
      subcloned into eukaryotic expression vector pcDNA3. The recombinant vector 
      pcDNA-CD151 was transfected into PK-15 cells and the transgenic cell line was 
      generated after G418 selection. Transcription of the CD151 cDNA in transgenic 
      cell line was detected by RT-PCR and immunofluorescence. The cell line, together 
      with control cell lines PK-15, 3D4-CD163 and MARC-145, was infected with PRRSV, 
      and the viral RNA genome or antigens in the infected cells was detected by RT-PCR 
      or immunofluorescence. At different time points post-infection, the virus was 
      harvested and titrated on MARC-145 cells. RESULTS: The expected size of porcine 
      CD151 cDNA was amplified with a sequence identity of 99.7% to the published 
      sequence. From the pcDNA-CD151-transfected cell culture, a transgenic cell line 
      PK15-CD151 was generated and correct expression of porcine CD151 was confirmed. 
      After PRRSV infection, the viral RNA genome and antigens were detected in the 
      cell-line. Although apparent cytopathic effect was not observed in the virally 
      infected cell line, the infectious virus with a high viral title was detected. 
      The cell line had been passed for more than 30 generations and no significant 
      difference in viral title was observed among generations 10, 20 and 30 after 
      PRRSV infection. CONCLUSION: Transfection of non-permissive PK-15 cells with 
      porcine CD151 cDNA conferred the susceptibility to PRRSV infection, indicating an 
      important role of the porcine CD151 in PRRSV infection of porcine cells.
FAU - Huang, Yanyan
AU  - Huang Y
AD  - College of Veterinary Medicine, Hangzhou University, Yangzhou 225009, China.
FAU - Guo, Rui
AU  - Guo R
FAU - Zhang, Yu
AU  - Zhang Y
FAU - Zhang, Xinyu
AU  - Zhang X
FAU - Xia, Xiaoli
AU  - Xia X
FAU - Sun, Huaichang
AU  - Sun H
LA  - chi
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - China
TA  - Wei Sheng Wu Xue Bao
JT  - Wei sheng wu xue bao = Acta microbiologica Sinica
JID - 21610860R
RN  - 0 (DNA, Complementary)
RN  - 0 (Tetraspanin 24)
RN  - 0 (Viral Envelope Proteins)
RN  - 0 (glycoprotein 5, PRRSV)
SB  - IM
MH  - Animals
MH  - Cell Line
MH  - Cloning, Molecular
MH  - DNA, Complementary/chemistry/genetics
MH  - Gene Expression
MH  - Genetic Predisposition to Disease
MH  - Porcine Reproductive and Respiratory Syndrome/genetics/virology
MH  - Porcine respiratory and reproductive syndrome virus/*physiology
MH  - Sequence Analysis, DNA
MH  - Swine
MH  - Tetraspanin 24/*genetics
MH  - Transcription, Genetic
MH  - Transgenes
MH  - Viral Envelope Proteins/genetics
MH  - Virus Replication
EDAT- 2013/08/21 06:00
MHDA- 2013/11/10 06:00
CRDT- 2013/08/21 06:00
PHST- 2013/08/21 06:00 [entrez]
PHST- 2013/08/21 06:00 [pubmed]
PHST- 2013/11/10 06:00 [medline]
PST - ppublish
SO  - Wei Sheng Wu Xue Bao. 2013 May 4;53(5):507-14.