Generating Recombinant Pseudorabies Virus for Use as a Vaccine Platform

Tan, Feifei; Li, Xiangdong; Tian, Kegong

doi:10.1007/978-1-4939-6869-5_5

Cited by 7 publications

(5 citation statements)

References 20 publications

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“…To construct PRV BAC clones with Fa gI/gE/11K or TK/gI/gE/11K genes replacement in HN1201genome, the plasmid pBAC-HN1201 TK− was used as the backbone. Firstly, a PCR fragment containing an I-SceI site together with the ampicillin (Amp) gene anked by a 50 bp duplicated sequence of HN1201 was ampli ed, and then be electroporated into the DY380 competent cells harboring pBAC-HN1201 TK− [21,24] to obtain the positive clones of pBAC-HN1201 TK−/gI−/gE−/11k− Amp, which were con rmed by digestion of BamHI. After that, a PCR fragment containing an I-SceI site together with the kanamycin (Kan) gene and the gI/gE/11k genes of Fa strain anked by a 500 bp sequence of HN1201 in each end was ampli ed, and then be transformed into E. coli DY380 competent cells to obtain the intermediate plasmid pBAC-HN1201 TK− -Fa gI+/gE+/11K+ -Kan. After digested with I-SceI, the linear plasmid was transformed into DY380 cells to remove the Kan gene, and the bacterial strain of pBAC-HN1201 TK− -Fa gI+/gE+/11K+ was obtained.…”

Section: Generation Of Bac Clonementioning

confidence: 99%

TK/gI/gE/11K Genes Is Not Involved in the Virulence Enhancement of Highly Virulent Pseudorabies Virus

Tan

Guo

Zhang

et al. 2021

Preprint

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The large-scale outbreaks of Pseudorabies in china since 2011 in vaccine-immunized pig farms were caused by the highly virulent Pseudorabies virus (PRV). To investigate the factors involved in the virulence enhancement of the variant PRV, four recombinant viruses with the virulence gene’s replacement, gI/gE/11K and TK/gI/gE/11K, between the variant strain HN1201 and the classical strain Fa were constructed based on bacterial artificial chromosome infectious clones. Compared with their parental strain, the viral titers of the recombinant PRV strains are not strongly influenced by the replacement of TK/gI/gE/11K, while as previously reported, the strain HN1201 and its derived viruses showed the higher mortality, the severer clinical symptoms and tissues damage than that of strain Fa and its derived strains. In summary, the TK/gI/gE/11K genes of variant strain HN1201 showed no contribute to its virulence enhancement compares with the classical strain.

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Section: Generation Of Bac Clonementioning

confidence: 99%

TK/gI/gE/11K Genes Is Not Involved in the Virulence Enhancement of Highly Virulent Pseudorabies Virus

Tan

Guo

Zhang

et al. 2021

Preprint

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“…The PRV genome encodes 16 envelope glycoproteins that function in viral entry, egress and cell-to-cell spread. The gE glycoprotein is the major virulence protein enabling PRV to invade the host nervous system [ 5 ]. Deletion of the gE gene significantly decreases virulence and prevents invasion of the trigeminal and olfactory nerve terminals [ 6 ].…”

Section: Introductionmentioning

confidence: 99%

Transcriptomic analysis reveals impact of gE/gI/TK deletions on host response to PRV infection

Wang,

Li,

Dong

et al. 2023

Virol J

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Background Pseudorabies virus (PRV) causes substantial losses in the swine industry worldwide. Attenuated PRV strains with deletions of immunomodulatory genes glycoprotein E (gE), glycoprotein I (gI) and thymidine kinase (TK) are candidate vaccines. However, the effects of gE/gI/TK deletions on PRV-host interactions are not well understood. Methods To characterize the impact of gE/gI/TK deletions on host cells, we analyzed and compared the transcriptomes of PK15 cells infected with wild-type PRV (SD2017), PRV with gE/gI/TK deletions (SD2017gE/gI/TK) using RNA-sequencing. Results The attenuated SD2017gE/gI/TK strain showed increased expression of inflammatory cytokines and pathways related to immunity compared to wild-type PRV. Cell cycle regulation and metabolic pathways were also perturbed. Conclusions Deletion of immunomodulatory genes altered PRV interactions with host cells and immune responses. This study provides insights into PRV vaccine design.

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“…Pseudorabies (PR), also called Aujeszky's disease, is caused by the infection of an alpha-herpesvirus Pseudorabies virus (PRV) (1). The double-stranded DNA genomic sequence of PRV is ∼145 kb in size, containing almost 70 open reading frames (ORFs) that encode 70-100 viral proteins (2).…”

Section: Introductionmentioning

confidence: 99%

Construction and Immunogenicity of a Recombinant Pseudorabies Virus Variant With TK/gI/gE/11k/28k Deletion

Yan¹,

Huang²,

Bai³

et al. 2022

Front. Vet. Sci.

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In China, the re-emerging pseudorabies virus (PRV) variant has caused large-scale outbreaks of pseudorabies in swine herds with classical PRV vaccine immunization since late 2011. Here, a recombinant PRV with TK/gI/gE/11k/28k deletion was constructed based on variant HN1201 strain isolated in 2012, by the bacterial artificial chromosome infectious clones. Compared with the parental virus, the recombinant PRV rHN1201TK−/gE−/gI−/11k−/28k− showed a similar virus grown curve and exhibited smaller plaques. The vaccination of rHN1201TK−/gE−/gI−/11k−/28k− could elicit an earlier and higher level of gB antibody, and the neutralizing antibodies elicited by rHN1201TK−/gE−/gI−/11k−/28k− were effective against both PRV classical and variant strains. Clinically, the body temperature of the pigs immunized with rHN1201TK−/gE−/gI−/11k−/28k− was significantly lower than that of the classical PRV vaccine immunized pigs, and the recombinant PRV could provide effective protection against the challenge with the PRV variant. These results imply that the rHN1201TK−/gE−/gI−/11k−/28k− could be a promising vaccine candidate for the prevention of the current epidemic of pseudorabies in China.

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Generating Recombinant Pseudorabies Virus for Use as a Vaccine Platform

Cited by 7 publications

References 20 publications

TK/gI/gE/11K Genes Is Not Involved in the Virulence Enhancement of Highly Virulent Pseudorabies Virus

TK/gI/gE/11K Genes Is Not Involved in the Virulence Enhancement of Highly Virulent Pseudorabies Virus

Transcriptomic analysis reveals impact of gE/gI/TK deletions on host response to PRV infection

Construction and Immunogenicity of a Recombinant Pseudorabies Virus Variant With TK/gI/gE/11k/28k Deletion

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