The construction and immunogenicity analyses of a recombinant pseudorabies virus with porcine circovirus type 3 capsid protein co-expression

2021 ◽  
pp. 109283
Author(s):  
Lun Yao ◽  
Yufang Cheng ◽  
Hao Wu ◽  
Ahmed. H. Ghonaim ◽  
Shengxian Fan ◽  
...  
Keyword(s):  
Capsid Protein ◽  
Pseudorabies Virus ◽  
Porcine Circovirus Type ◽  
Porcine Circovirus ◽  
Recombinant Pseudorabies Virus ◽  
Type 3

scholarly journals Development of an indirect immunofluorescence assay for PCV3 antibody detection based on capsid protein

2021 ◽  
Vol 1 (1) ◽  
Author(s):  
Lun Yao ◽  
Chang Li ◽  
Junwei Wang ◽  
Yufang Cheng ◽  
Ahmed H. Ghonaim ◽  
...  
Keyword(s):  
Protein Expression ◽  
Capsid Protein ◽  
Nuclear Localization ◽  
Pseudorabies Virus ◽  
Expression System ◽  
Porcine Circovirus Type ◽  
Immunofluorescence Assay ◽  
Antibody Detection ◽  
Porcine Circovirus ◽  
Transfected Cells

AbstractPorcine circovirus type 3 (PCV3) is a novel porcine circovirus associated with porcine dermatitis and nephritis syndrome (PDNS), reproductive failure, and multisystemic inflammation. Capsid protein (Cap) encoded by PCV3 ORF2 gene has been identified as an immunogenic protein. Currently, there is no immunofluorescence assay (IFA) available for serological diagnosis. Here, the N-terminal 33 amino acids of Cap protein were predicted to serve as a PCV3 nuclear localization signal (NLS). Two types of recombinant plasmids were constructed for recombinant protein expression in Sf9 cells by using a baculovirus expression system: plasmid rvBac-Pc for full-length Cap protein expression and rvBac-Sc for Cap protein expression with a honeybee melittin signal peptide in place of the predicted NLS sequence. Expression of the nuclear localization sequences was further analyzed by IFA. Strong and specific fluorescence signals were observed in the nucleus of rvBac-Pc-transfected cells and in the cytoplasm of rvBac-Sc-transfected cells. No cross-reactivity was observed with porcine circovirus type 2, porcine pseudorabies virus, classical swine fever virus, or porcine reproductive and respiratory syndrome virus. In summary, we developed two fluorescence detection modes for Cap protein that can be used to detect PCV3 antibodies. This method is suitable  for the diagnosis and epidemiological investigation of PCV3. This study provides a reliable detection method for monitoring PCV3 antibody level in pigs in the future.

Generation and immunogenicity of a recombinant pseudorabies virus expressing cap protein of porcine circovirus type 2

2007 ◽  
Vol 119 (2-4) ◽  
pp. 97-104 ◽  
Author(s):  
Yunfeng Song ◽  
Meilin Jin ◽  
Songlin Zhang ◽  
Xiaojuan Xu ◽  
Shaobo Xiao ◽  
...  
Keyword(s):  
Pseudorabies Virus ◽  
Porcine Circovirus Type ◽  
Porcine Circovirus Type 2 ◽  
Porcine Circovirus ◽  
Recombinant Pseudorabies Virus

scholarly journals Structural insight into the type-specific epitope of porcine circovirus type 3

2020 ◽  
Vol 40 (6) ◽  
Author(s):  
Mingfang Bi ◽  
Xiangdong Li ◽  
Weifeng Zhai ◽  
Bo Yin ◽  
Kegong Tian ◽  
...  
Keyword(s):  
Capsid Protein ◽  
Self Assembly ◽  
Structural Information ◽  
Porcine Circovirus Type ◽  
Porcine Circovirus ◽  
Icosahedral Symmetry ◽  
Western Blot Assay ◽  
Structural Framework ◽  
Loop Region ◽  
Type 3

Abstract The recently identified pathogenic Porcine circovirus type 3 (PCV3) may threaten to reduce the pig population dramatically worldwide. In our previous study, a PCV3-specific monoclonal antibody (mAb-1H11) was successfully applied in immune-histochemistry staining and ELISA, which specifically recognize PCV3 capsid protein in PCV3-positive pig tissues. In the present study, we expressed and purified the soluble sole capsid protein of PCV3. The purified capsid protein was capable of self-assembly into virus-like-particles (VLPs), which is validated by transmission electron microscopy and dynamic light scattering assays. Moreover, the epitope of mAb-1H11 was identified in the CD-loop region (a.a. 72-79) on the VLP surface, which is confirmed by PCV2-PCV3 epitope swapping assay. For the first time, we determined the cryo-EM structure of PCV3-VLP at 8.5 Å resolution that reveals the detailed structural information of PCV3-VLP. In our cryo-EM structure, PCV3-VLP is composed of 60 capsid protein subunits assembled with T = 1 icosahedral symmetry. Consistent to our bio-dot Western blot assay, the structural comparison between PCV3 and PCV2 revealed significant structural differences in the surface-exposed loops, including the CD-loop (a.a. 72-79) and the EF-loop (a.a. 109-131). Our work provides a structural framework for engineering future PCV3 vaccine and diagnosis kits development.

scholarly journals High repetitive arginine in the anterior of PCV3 capsid protein is a severe obstacle for its expression in E. coli

AMB Express ◽  
2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Bing Yan Liu ◽  
Bin Gao ◽  
Meng Zhi Liu ◽  
Ting Ting Zhang ◽  
Bao Shan Liu ◽  
...  
Keyword(s):  
Capsid Protein ◽  
Vaccine Candidate ◽  
Clinical Signs ◽  
Porcine Circovirus Type ◽  
Reproductive Failure ◽  
Porcine Circovirus ◽  
Effective Vaccine ◽  
E Coli ◽  
Efficient Expression ◽  
Type 3

AbstractPorcine circovirus type 3 (PCV3) is a novel circovirus identified in sows with PDNS-like clinical signs and reproductive failure. The capsid protein (CAP) of PCV3 is expected to be an effective vaccine candidate. Here, we expressed the original capsid protein, truncated capsid protein without anterior highly repetitive arginine (ΔCAP) and their codon-optimized counterparts in E. coli. These results showed that lots of repeated arginine could severely lower the expression of PCV3 capsid protein in E. coli. At the same time, the recombined truncated PCV3 capsid protein forms typic virions. The efficient expression of capsid protein is expected to serve the development of PCV3 vaccines and other studies of PCV3 capsid protein.

scholarly journals The serine-48 residue of nucleolar phosphoprotein nucleophosmin-1 plays critical role in subcellular localization and interaction with porcine circovirus type 3 capsid protein

2021 ◽  
Vol 52 (1) ◽  
Author(s):  
Jianwei Zhou ◽  
Juan Li ◽  
Haimin Li ◽  
Ying Zhang ◽  
Weiren Dong ◽  
...  
Keyword(s):  
Capsid Protein ◽  
Critical Role ◽  
Porcine Circovirus Type ◽  
Porcine Circovirus ◽  
Nucleolar Localization ◽  
Nucleolar Localization Signal ◽  
Localization Signal ◽  
Charge Property ◽  
Type 3 ◽  
First Time

AbstractThe transport of circovirus capsid protein into nucleus is essential for viral replication in infected cell. However, the role of nucleolar shuttle proteins during porcine circovirus 3 capsid protein (PCV3 Cap) import is still not understood. Here, we report a previously unidentified nucleolar localization signal (NoLS) of PCV3 Cap, which hijacks the nucleolar phosphoprotein nucleophosmin-1 (NPM1) to facilitate nucleolar localization of PCV3 Cap. The NoLS of PCV3 Cap and serine-48 residue of N-terminal oligomerization domain of NPM1 are essential for PCV3 Cap/NPM1 interaction. In addition, charge property of serine-48 residue of NPM1 is critical for nucleolar localization and interaction with PCV3 Cap. Taken together, our findings demonstrate for the first time that NPM1 interacts with PCV3 Cap and is responsible for its nucleolar localization.

Sign in / Sign up

Export Citation Format

Share Document