Construction and immunogenicity of a gE/gI/TK-deleted PRV based on porcine pseudorabies virus variant

Zhao, Yu; Wang, Linqing; Zheng, Huihua; Yang, Yurong; Liu, Fang; Zheng, Lan-Lan; Jin, Yue; Chen, Hongying

doi:10.1016/j.mcp.2020.101605

Cited by 42 publications

(27 citation statements)

References 37 publications

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“…One hundred seventy six-week-old healthy BALB/c female mice were randomly divided into 17 groups of 10. Mice in PRV-injected groups were inoculated subcutaneously (s.c.) with the different isolates of a virus titer of 10 5.0 50% tissue culture infectious doses (TCID 50 )/dose as previously described, respectively ( 21 ). Mice in the DMEM-injected group were inoculated with the same dose of DMEM as control.…”

Section: Methodsmentioning

confidence: 99%

“…The complete gE, gC, and gB genes of PRV were amplified from viral DNA extracted from PRV isolates using specific primers gE-F/R, gC-F/R, and gB-F/R designed in our laboratory (21). The complete gD gene was amplified by PCR from viral DNA using a pair of primers gD-F/R (gD-F: 5 ′ -ATCCACTCCCAGCGGTCCACAAAAT−3 ′ , gD-R: 5 ′ -AAAAACAGCAGCGTCCCGTCTATCG−3 ′ ) with the same PCR volume mixture as that of the partial gE gene, and the cycling protocol was as follows: an initial denaturation at 95 • C for 5 min, 35 cycles of denaturation at 95 • C for 55 s, annealing at 57 • C for 1 min, extension at 72 • C for 100 s, and a final extension at 72 • C for 10 min.…”

Section: Sequencing and Phylogenetic Analysismentioning

confidence: 99%

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Isolation and Phylogenetic Analysis of Reemerging Pseudorabies Virus Within Pig Populations in Central China During 2012 to 2019

Zheng

Bai

et al. 2021

Front. Vet. Sci.

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To understand the biological characteristics of the reemerging pseudorabies virus (PRV) strains, a total of 392 tissue samples were collected from diseased pigs during reemerging PR outbreaks between 2012 and 2019 on farms in central China where swine had been immunized with Bartha-K61 and 51 (13. 01%) were positive for the gE gene by PCR. Sixteen PRV strains were isolated and caused clinical symptoms and death in mice. Subsequently, gE, gC, gB, and gD complete genes were amplified from the 16 PRV isolates and sequenced. Phylogenetic analysis based on these four gene sequences shows that the 16 PRV isolates were more closely related to the Chinese PRV variants (after 2012) but genetically differed from early Chinese PRV isolates (before 2012). Sequence analysis reveals that PRV isolates exhibited amino acid insertions, substitutions, or deletions compared with early Chinese PRV isolates and European–American PRV strains. In addition, this is the first report that eight isolates (8/16) in this study harbor a unique amino acid substitution at position 280 (F to L) of the gC protein, and six isolates have an amino acid substitution at position 338 (A to V) of the gD protein compared with the Chinese PRV variants. The emulsion containing inactivated PRV NY isolate could provide complete protection against the NY isolate. This study might enrich our understanding of the evolution of reemerging PRV strains as well as pave the way for finding a model virus to develop a novel vaccine based on reemerging PRV strains.

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Section: Methodsmentioning

confidence: 99%

Section: Sequencing and Phylogenetic Analysismentioning

confidence: 99%

Isolation and Phylogenetic Analysis of Reemerging Pseudorabies Virus Within Pig Populations in Central China During 2012 to 2019

Zheng

Bai

et al. 2021

Front. Vet. Sci.

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“…Similarly, a triple gE/gI/TK gene-inactivated HeN1 PRV strain has been demonstrated to be fully attenuated and can provide immune protection against parental PRV challenge [ 109 ]. Based on a PRV variant (NY strain), a triple gE/gI/TK-deleted mutant has been recently constructed through homologous DNA recombination and CRISPR/Cas9-based gene-editing technology, providing a next-generation vaccine candidate for control of potential new variant PRV strains [ 110 ]. Using the CRISPR/Cas9 system, knockout of non-essential gene 8-DR from the genome of ASFV virulent strain Georgia07 in porcine macrophages was also reported to successfully rescue the recombinant virus, which laid a foundation for the follow-up vaccine development for future control of African swine fever [ 111 ].…”

Section: Crispr/cas System In Virology Researchmentioning

confidence: 99%

Latest Advances of Virology Research Using CRISPR/Cas9-Based Gene-Editing Technology and Its Application to Vaccine Development

Teng

Yao

Nair

et al. 2021

Viruses

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In recent years, the CRISPR/Cas9-based gene-editing techniques have been well developed and applied widely in several aspects of research in the biological sciences, in many species, including humans, animals, plants, and even in viruses. Modification of the viral genome is crucial for revealing gene function, virus pathogenesis, gene therapy, genetic engineering, and vaccine development. Herein, we have provided a brief review of the different technologies for the modification of the viral genomes. Particularly, we have focused on the recently developed CRISPR/Cas9-based gene-editing system, detailing its origin, functional principles, and touching on its latest achievements in virology research and applications in vaccine development, especially in large DNA viruses of humans and animals. Future prospects of CRISPR/Cas9-based gene-editing technology in virology research, including the potential shortcomings, are also discussed.

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“…The complete gE and gC genes of PRV were amplified from viral DNA extracted from PRV isolates using two specific primers gE-F/R and gC-F/R designed in our laboratory (Zhao et al, 2020). Then the genes were ligated with the vector pMD18-T (Takara), and separately introduced toEscherichia coli DH-5a cells (Takara).…”

Section: Sequencing and Phylogenetic Analysis Of Ge And Gc Genesmentioning

confidence: 99%

Molecular epidemiology and biological characteristics of swine pseudorabies virus in Henan province of China during 2012 to 2019

Zheng¹,

Bai²,

Zhao³

et al. 2020

Preprint

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Since late 2011, pseudorabies virus (PRV; Suid herpesvirus 1) infection was widely prevalent in vaccinated swine farms in China, and caused tremendous economic losses in the swine industry. To understand the epidemic and biological characteristics of the virus, a total of 1,174 tissue samples were collected from Bartha-K61-immunized swine farms in Henan province of China between 2012 and 2019, and PRV strains were isolated and the complete sequences of gE and gC genes were amplified by PCR. The detection rate of PRV was 15.25% (179/1174), which varied from 6.61% to 25.00% between 2012 and 2019. And 16 PRV isolates were obtained, and could cause clinical symptoms and death in mice. The phylogenetic trees based on the sequences of gE and gC genes showed that the 16 PRV strains in this study at these two phylogenetic trees all clustered to a relatively independent branch altogether with the Chinese variant PRV strains (after 2012), and sequence analysis of the isolates revealed that gE and gC both contained amino acid insertions, substitutions or deletions compared with European-American PRV strains and early Chinese PRV strains (before 2012). In addition, it was the first report that eight strains (8/16) in this study harbored a unique amino acid substitution at site 280 (F to L) of gC gene. In the protection assay, the emulsion containing inactivated PRV NY isolate could provide complete protection against variant NY, and the titer of neutralizing antibodies was 1:82. This study might enrich our understanding of the evolution of variant PRVs as well as pave the way for finding a model virus to develop a novel vaccine based on PRV variants.

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Construction and immunogenicity of a gE/gI/TK-deleted PRV based on porcine pseudorabies virus variant

Cited by 42 publications

References 37 publications

Isolation and Phylogenetic Analysis of Reemerging Pseudorabies Virus Within Pig Populations in Central China During 2012 to 2019

Isolation and Phylogenetic Analysis of Reemerging Pseudorabies Virus Within Pig Populations in Central China During 2012 to 2019

Latest Advances of Virology Research Using CRISPR/Cas9-Based Gene-Editing Technology and Its Application to Vaccine Development

Molecular epidemiology and biological characteristics of swine pseudorabies virus in Henan province of China during 2012 to 2019

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