PMID- 35126342
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20220208
IS  - 1664-302X (Print)
IS  - 1664-302X (Electronic)
IS  - 1664-302X (Linking)
VI  - 13
DP  - 2022
TI  - A Recombinant Porcine Reproductive and Respiratory Syndrome Virus Stably 
      Expressing DsRed Protein Based on Bacterial Artificial Chromosome System.
PG  - 839845
LID - 10.3389/fmicb.2022.839845 [doi]
LID - 839845
AB  - Recombinant viruses possessing reporter proteins as tools are widely applied in 
      investigating viral biology because of the convenience for observation. 
      Previously, we generated a recombinant pathogenic porcine reproductive and 
      respiratory syndrome virus (PRRSV) with enhanced green fluorescent protein (EGFP) 
      reporter for monitoring virus spread and screening of neutralizing antibodies. 
      PRRSV with different kinds of reporters can support more application scenarios. 
      Here, we described a new genetically stable infectious clones of a highly 
      pathogenic PRRSV (HP-PRRSV) harboring the DsRed (a red fluorescent protein 
      isolated from the coral Discosoma) gene. In the recombinant infectious clone, the 
      transcription regulatory sequence 2 (TRS2) of PRRSV was inserted between the open 
      reading frame 7 (ORF7) and 3'UTR to drive the transcription of DsRed gene, which 
      makes it a separate transcription unit in the viral genome. Using the bacterial 
      artificial chromosome (BAC) system and cytomegalovirus (CMV) promoter, the 
      recombinant HP-PRRSV with the DsRed insertion was successfully rescued and showed 
      similar growth and replication patterns compared with the wild-type virus in the 
      MARC-145 cells. In addition, the DsRed protein was stably expressed in the 
      recombinant virus for at least 10 passages with consistent fluorescence intensity 
      and density. Using the recombinant HP-PRRSV with DsRed protein, the virus 
      tracking in MARC-145 was observed by live-cell imaging. Meanwhile, quantification 
      of the DsRed fluorescence positive cells by flow cytometry provides an 
      alternative to standard methods for testing the level of PRRSV infection. This 
      recombinant PRRSV with DsRed fluorescence protein expression could be a useful 
      tool for fundamental research on the viral biology and shows the new design for 
      stable expression of foreign genes in PRRSV.
CI  - Copyright (c) 2022 Li, Zhang, Yao, Shi, Zhao, Huang and Sun.
FAU - Li, Na
AU  - Li N
AD  - Key Laboratory of Ecological Security, Collaborative Innovation Centre of Water 
      Security for Water Source Region of Mid-line of South-to-North Diversion Project 
      of Henan Province, Henan Provincial Engineering and Technology Center of Health 
      Products for Livestock and Poultry, School of Life Sciences and Agricultural 
      Engineering, Nanyang Normal University, Nanyang, China.
FAU - Zhang, Yiyi
AU  - Zhang Y
AD  - Key Laboratory of Ecological Security, Collaborative Innovation Centre of Water 
      Security for Water Source Region of Mid-line of South-to-North Diversion Project 
      of Henan Province, Henan Provincial Engineering and Technology Center of Health 
      Products for Livestock and Poultry, School of Life Sciences and Agricultural 
      Engineering, Nanyang Normal University, Nanyang, China.
FAU - Yao, Lunguang
AU  - Yao L
AD  - Key Laboratory of Ecological Security, Collaborative Innovation Centre of Water 
      Security for Water Source Region of Mid-line of South-to-North Diversion Project 
      of Henan Province, Henan Provincial Engineering and Technology Center of Health 
      Products for Livestock and Poultry, School of Life Sciences and Agricultural 
      Engineering, Nanyang Normal University, Nanyang, China.
FAU - Shi, Yunpeng
AU  - Shi Y
AD  - Shijiazhuang Customs (Huanghua Port), Cangzhou, China.
FAU - Zhao, Qin
AU  - Zhao Q
AD  - College of Veterinary Medicine, Northwest A&F University, Yangling, China.
FAU - Huang, Baicheng
AU  - Huang B
AD  - National Research Center for Veterinary Medicine, Luoyang, China.
FAU - Sun, Yani
AU  - Sun Y
AD  - College of Veterinary Medicine, Northwest A&F University, Yangling, China.
LA  - eng
PT  - Journal Article
DEP - 20220121
PL  - Switzerland
TA  - Front Microbiol
JT  - Frontiers in microbiology
JID - 101548977
PMC - PMC8814527
OTO - NOTNLM
OT  - DsRed
OT  - HP-PRRSV
OT  - bacterial artificial chromosome
OT  - reporter
OT  - transcription regulatory sequence
COIS- YuS was employed by Shijiazhuang Customs (Huanghua Port). The remaining authors 
      declare that the research was conducted in the absence of any commercial or 
      financial relationships that could be construed as a potential conflict of 
      interest.
EDAT- 2022/02/08 06:00
MHDA- 2022/02/08 06:01
PMCR- 2022/01/21
CRDT- 2022/02/07 05:30
PHST- 2021/12/20 00:00 [received]
PHST- 2022/01/03 00:00 [accepted]
PHST- 2022/02/07 05:30 [entrez]
PHST- 2022/02/08 06:00 [pubmed]
PHST- 2022/02/08 06:01 [medline]
PHST- 2022/01/21 00:00 [pmc-release]
AID - 10.3389/fmicb.2022.839845 [doi]
PST - epublish
SO  - Front Microbiol. 2022 Jan 21;13:839845. doi: 10.3389/fmicb.2022.839845. 
      eCollection 2022.