• Title/Summary/Keyword: Pseudorabies virus

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A Restrictive Virus Tropism, Latency and Reactivation of Pseudorabies Virus Following Irreversible Deletion of Bsrl Restriction Site in the Thymidine-kinase Gene

  • Mohd Lila Mohd Azmi;Zeenathul, Nazariah-Allaudin;Abdel-Wahid Saeed Ali;Che Abdul Rahim Mohamed;Kamarudin, Awag-Isa
    • Journal of Microbiology
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    • v.40 no.1
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    • pp.1-10
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    • 2002
  • At the dose of 1000 p.f.u. per mouse,100% mortality occurred in mice inoculated with wild-type pseudorabies virus (PrV). In contrast, upon stable deletion of 10 bp nucleotides at the Bsrl site within the TK gene, PrV was rendered to be completely apathogenic. The deletion also caused the virus to be less capable of replicating in respiratory as well as in nervous system tissues. Although animals were exposed to high titers of TK-deleted PrVs, the virus failed to replicate to a high titer as compared to the pathogenic parental virus. In contrast to previous studies the deletion in the TK gene did not prevent the virus from establishing latency. Upon immunosuppression, the latent virus? however, reactivated but replicated at low titers. Interestingly, TK-deleted virus established latency and reactivation, that are occurred only in trigeminal ganglia and the cerebrums and no other tissues involved. Following reactivation, there was no indication of virus shedding in respiratory tissues as confirmed by virus isolation and polymerase chain reaction (PCR) technique targeting at the gB gene of PrV, The non-pathogenic virus with non-shedding characteristics, upon reactivation of the latent virus, would be the important feature of a live virus vaccine candidate.

Expression of Pseudorabies Virus (PRV) Glycoproteins gB, gC and gD using Bacterial Expression System

  • Yun, Bit-Na-Rae;Bae, Sung-Min;Lee, Jun-Beom;Kim, Hee-Jung;Woo, Soo-Dong
    • International Journal of Industrial Entomology and Biomaterials
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    • v.23 no.1
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    • pp.147-153
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    • 2011
  • The Pseudorabies (PR), also called Aujeszky's disease (AD), is an infectious viral disease caused by an alpha herpes virus and has domestic and wild pigs, as well as a wide range of domestic and wild animals, as the natural host. Pseudorabies virus (PRV) virions contain several envelope glycoproteins. Among them, gB, gC and gD are regarded as the major immunogenic proteins. We expressed these glycoproteins using the bacterial expression system and analyzed recombinant proteins. Expression of glycoproteins gC and gD were observed on SDS-PAGE or Western blot analysis, but gB was not. Optimal concentration of IPTG and inducing time were determined as 1.0 mM and 4 h, respectively, for the expression of both gC and gD in E. coli. A sodium dodecyl sulfate (SDS) was the most efficient detergent in solubilizing insoluble recombinant protein.

Application of Monoclonal Antibody to Develop Diagnostic Techniques for Infectious Bovine Rhinotracheitis Virus I. Production of Monoclonal Antibodies against Infectious Bovine Rhinotracheitis Virus (단(單)클론성 항체(抗體)를 이용한 소전염성비기관염(傳染性鼻氣管炎)바이러스 진단법(診斷法) 개발 I. 소전염성비기관염(傳染性鼻氣管炎)바이러스에 대한 단(單)클론성 항체(抗體) 생산(生産))

  • Jun, Moo Hyung;Kim, Duck Hwan;Lee, Hun Jun;An, Soo Hwan;Kweon, Chang Hee
    • Korean Journal of Agricultural Science
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    • v.14 no.2
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    • pp.401-408
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    • 1987
  • Nine monoclonal antibodies directed against infectious bovine rhinotracheitis virus (IBRV) were prepared by using cell hybridization technique, and the biological properties of the antibodies were investigated by means of immunofluorescence, serum neutralization, and electrophoretic analysis. Eight of 9 monoclonal antibodies reacted specifically with the antigenic constituents of IBRV, infectious laryngotracheitis virus, Marek's disease virus, turkey herpesvirus, hog cholera virus, porcine parvovirus and transmissible gastroenteritis virus. However, the remaining one, 26-2 clone, was found to be cross-reactive with pseudorabies virus. Two monoclonal antibodies, 7-C-2 and 12-A-2, which had neutralizing activity, were reactive with the molecular weights of 72 kilo daltons (72K) and 125K of IBRV proteins electrophoretically separated, respectively. The monoclonal antibody, 3-H-3, which is corresponding to 94K of IBRV proteins, revealed no neutralizing activity. The cross-reactive monoclonal antibody, 26-2, was proved by electrophoretical analysis to be reactive with 100K of IBRV proteins and 40K of pseudorabies virus.

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Pathological Studies on Pseudorabies(Aujeszky's Disease) I. Histopathological Findings of Experimentally Infected Pigs (가성광견병에 관한 병리학적 연구 I. 인공감염돈의 병리조직학적 소견)

  • Kim Soon-bok;Kwak Soo-dong;Wittman Georg;Olinger Volker
    • Journal of the korean veterinary medical association
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    • v.22 no.5
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    • pp.294-298
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    • 1986
  • In pigs inoculated with Pseudorabies virus, clinical, anatomical and histopathological findings were observed. Nervous sings, high fever and collapse were main clinical symptoms. Anatomical findings were not prominent except swollen lymph nodes with hemor

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Cloning of Major Capsid Protein Gene of Pseudorabies Virus and Expression by Baculovirus Vector System (Pseudorabies Virus의 Major Capsid Protein 유전자의 클론닝과 Baculovirus Vector System에 의한 발현)

  • An, Dong-Jun;Jun, Moo-Hyung;Song, Jae-Young;Park, Jong-Hyeon;Hyun, Bang-Hun;Chang, Kyung-Soo;An, Soo-Hwan
    • The Journal of Korean Society of Virology
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    • v.26 no.2
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    • pp.151-162
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    • 1996
  • Pseudorabies is caused by Pseudorabies virus (PRV: Aujeszky's disease virus) of Herpesviridae that is characterized by 100 to 150nm in size with a linear double-stranded DNA molecule with of approximately $90{\times}10^6Da$. This disease affects most of domestic animals such as swine, cattle, dog, sheep, cat, chicken, etc. causing high mortality and economic losses. In swine, young piglets show high mortality and pregnant sows, reproductive failures. However the adult swine reveals no clinical signs in general. But they become a carrier state and play an important role for propagation of the disease. In this study, the nucleotide sequence of major casid protein gene of PRV, Yangsan strain isolated from the diseased swine in Korea was analyzed, and the recombinant MCP was produced by expression of the MCP gene in Sf-9 cell using baculovirus transfer vector system. As result, in BamHI digestion, MCP gene locus of PRV YS strain showed different from that of Indiana S strain. The patterns of enzyme mapping were also found to be unidentical each other. The sequence of the MCP gene partially analyzed showed 98.09% identity to Indiana S strain. The expression of MCP in Sf-9 cell cotransfected by pVLMCP-44 baculovirus expression vector was characterized by Southern blot hybridization, immunofluoresent and immunocytochemical tests, SDS-PAGE and Western blotting. The rMCP with M.W. 142kDa was most effectively expressed in Sf-9 cells at the 3-4th days post inoculation of the recombinant baculovirus by 2 moi.

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Epidemiological investigation of porcine pseudorabies virus and its coinfection rate in Shandong Province in China from 2015 to 2018

  • Ma, Zicheng;Han, Zifeng;Liu, Zhaohu;Meng, Fanliang;Wang, Hongyu;Cao, Longlong;Li, Yan;Jiao, Qiulin;Liu, Sidang;Liu, Mengda
    • Journal of Veterinary Science
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    • v.21 no.3
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    • pp.36.1-36.9
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    • 2020
  • Background: Pseudorabies, also known as Aujeszky's disease, is caused by the pseudorabies virus (PRV) and has been recognized as a critical disease affecting the pig industry and a wide range of animals around the world, resulting in great economic losses each year. Shandong province, one of the most vital food animal-breeding regions in China, has a very dense pig population, within which pseudorabies infections were detected in recent years. The data, however, on PRV epidemiology and coinfection rates of PRV with other major swine diseases is sparse. Objectives: This study aimed to investigate the PRV epidemiology in Shandong and analyze the current control measures. Methods: In this study, a total number of 16,457 serum samples and 1,638 tissue samples, which were collected from 362 intensive pig farms (≥ 300 sows/farm) covered all cities in Shandong, were tested by performing enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR). Results: Overall, 52.7% and 91.5% of the serum samples were positive for PRV-gE and -gB, respectively, based on ELISA results. In addition, 15.7% of the tissue samples were PCR positive for PRV. The coinfection rates of PRV with porcine circovirus type 2 (PCV2), porcine reproductive and respiratory syndrome virus, and classical swine fever virus were measured; coinfection with PCV2 was 35.0%, higher than those of the other two viruses. Macroscopic and microscopic lesions were observed in various tissues during histopathological examination. Conclusions: The results demonstrate the PRV prevalence and its coinfection rates in Shandong province and indicate that pseudorabies is endemic in pig farms in this region. This study provides epidemiological data that can be useful in the prevention and control of pseudorabies in Shandong, China.

Immunogenicity of the Recombinant Pseudorabies Virus Major Capsid Protein Expressed by Baculovirus Vector System (Baculovirus Vector System에 의해 발현된 재조합 Pseudorabies Virus Major Capsid Protein의 면역원성)

  • Jun, Moo-Hyung;An, Dong-Jun;Chang, Kyung-Soo;Cho, Young-Sung;Park, Jong-Hyeon;Song, Jae-Young;Hyun, Bang-Hun;An, Soo-Hwan
    • The Journal of Korean Society of Virology
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    • v.26 no.2
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    • pp.163-171
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    • 1996
  • The recombinant pseudorabies virus major capsid protein (rMCP) was produced by expression of the MCP gene in Sf-9 cell using baculovirus transfer vector system. Following evaluation of the immunochemical properties of the rMCP, the immunogenicity of the recombinant subunit protiens were investigated in guinea pig and swine to obtain the preliminary guide line for the subunit vaccine using rMCP and gP50. It was proved that ultrasonication and 30% ammonium sulfate was most efficient to concentrate and purify the protein. The rMCP was safe in mice, guinea pigs and piglets. In guinea pigs, rMCP mixed with various adjuvants induced substantial degree of serum neutralizing antibody titers, but revealed incomplete protectivity against challenge. In swine, the combination of rMCP and gP50 showed the higher serum neutralizing antibody titers and cellular immune responses than rMCP alone. However, the protectivity was lower in comparison with the commercial gI-deleted inactivated vaccine. We expect these results to contribute to characterization of MCP gene of Korean isolate of PRV and to ultilize as preliminary information for prodution and evaluation of PRV recombinant subunit vaccines.

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Studies on the cloning gp50 and gp63 genes of Pseudorabies virus(Shope strain) (Pseudorabies virus의 gp50과 gp63 유전자 클로닝에 관한 연구)

  • Kweon, Chang-hee;Song, Jae-young;Kim, Byoung-han;Lee, Jung-bok;Lee, jae-chin;An, Soo-hwan;Lee, Yong-soon;Susumu, Maeda
    • Korean Journal of Veterinary Research
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    • v.31 no.3
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    • pp.311-318
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    • 1991
  • The DNA fragment representing for Pseudorabies gp50 and gp60(Shope) was cloned by recombinant techniques. The viral DNA was extracted from the infected cells and digested with Bam HI. The 6.8 Kb of Bam HI fragment was isolated from agarose gel and further digested with Nde I followed by Klenow treatment. The blunt ended 4.9Kb fragment was cloned into pTZ18R plasmid vector. The upstream region of gp50 was further manipulated to remove its 5' promoter region and create EcoRl site for possible eukaryotic expression system. The result of partial sequencing of cloned DNA indicated that Shope strain showed 95% homology with gp50 of Rice strain.

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Expression of Pseudorabies Virus Glycoproteins gB, gC and gD using Insect Expression System

  • Yun, Bit Na Rae;Gwak, Won Seok;Lee, Ji Hoon;Woo, Soo Dong
    • International Journal of Industrial Entomology and Biomaterials
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    • v.35 no.2
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    • pp.118-122
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    • 2017
  • Porcine pseudorabies virus (PRV) causes the Aujeszky's disease (AD) which is economically important disease in the swine industry worldwide. Killed or live vaccines have been used to control this disease, but their efficacy and side effects remain problems to be solved. To solve these problems, in this study, production of recombinant PRV glycoprotein gB, gC and gD was investigated in insect expression system. Glycoprotein gB, gC and gD are regarded as the major immunogenic antigens in PRV. Abundant production and immunogenicity of glycoprotein gB, gC and gD were confirmed by SDS-PAGE and Western blot analysis, respectively. Optimal infection dose and time were also determined for the production of each recombinant PRV glycoprotein. Confirmation of glycosylation of recombinant gB, gC and gD suggested their usefulness as antigens for the development of diagnosis kit or vaccines for Aujeszky's disease.