• Title/Summary/Keyword: capsid protein

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Immunohistochemical diagnosis of infectious pancreatic necrosis (어류 전염성훼장괴사증의 면역조직화학적 진단)

  • Kim, Soon-Bok
    • Korean Journal of Veterinary Pathology
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    • v.3 no.1
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    • pp.1-5
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    • 1999
  • This experiment was carried out to establish the immunohistochemical diagnostic method for infectious pancreatic necrosis in the monolayers of CHSE-214 cell cultures and paraffin-embedded tissue sections from rainbow trout infected with infectious pancreatic necrosis virus(IPNV). Specific identification of IPNV antigens was often demonstrated in the pancreatic exocrine cells, and less in the intestinal mucous epithelia and the renal hemopoietic tissues by the use of monoclonal antibodies against capsid protein VP2. The specific reaction was seen as a distinct red cytoplasmic color, often as small granules of various sizes. The result showed that streptavidin alkaline phosphatase immunohistochemisry specifically identified IPNV antigens in both infected cell cultures and tissue sections.

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Purification of Odontoglossum Ringspot Virus by DEAE-Cellulose Chromatography (DEAE 셀루로오즈 컬럼 크로마토그래피 기법에 의한 Odontoglossum 윤문 바이러스의 정제)

  • 이철호;박종오;정효원;나용준
    • Korean Journal Plant Pathology
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    • v.14 no.6
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    • pp.559-562
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    • 1998
  • Odontoglossum ringspot virus (ORSV) was finally purified from ORSV-infected orchid plants by diethylaminoethyl (DEAE) cellulose anion exchange column chromatography. The virus was reliably eluted by potassium chloride at the concentration from 0.1 M to 0.13 M. Partial purification was done by solubilization with Triton X-100 (allkylphenoxypolyethoxy ethanol) and precipitation with polyethylene glycol (PEG; MW 8,000). The finally purified ORSV represented one distinct homogeneous band and the molecular weight of its capsid protein was about 17,500 Dalton in electrophoretic analysis. Electron microscopy showed not only intact particles ranged from 280 nm to 340 nm in length, but also segmented particles that final 140 nm to 220 nm and even disks. Enzyme-linked immunosorbent assay (ELISA) showed that final yield was 12 mg/100 g of the infected leaves. Bioassay demonstrated that the purified ORSV had the normal infectivity to orchid plants and Nicotiana glutionsa. Based on these data, anion exchange column chromatography could be efficiently applied to the purification of ORSV and other viruses similar to ORSV.

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Infectious RNA Viruses in the Edible Mushroom Pleurotus spp.

  • Park, Jeonga-Soo;Kim, Young-Ho
    • Journal of Microbiology
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    • v.34 no.1
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    • pp.61-67
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    • 1996
  • Double-stranded RNA (dsRNA) viruses and single-stranded RNA(ssRNA) viruses were detected in a strain of Pleurotus mushroom cultivated in a farm. Those fungal virsus were purified in the pH 6.0 or pH 7.2 using CsCI or Cs$_{2}$SO$_{4}$ buoyant density centrifugation. Each viral particles were not completely separated at any trials. However, mushroom bacili-form virus contains a single major nucleic acid with 0.7 Kb ssRNA, which might code for 20 Kd viral capsid protein. The dsRNAs are encapsidatred into spherical-form viruses, whereas ssRNA viral genomes are encapsidated into two different sizes of bacili-form particles. A healthy-looking mushroom also contained some spherical-form viruses with dsRNAs. Laboratory strains of Pleurotus ostreatus and a cultivated strain of P. sajor-caju did not show any viral particles. Mushrooms with specific disease symptoms. however, contained at least four different sizes of spherical-form viruses. Thus, we concluded that a bacilli-form virus case a severe disease symptoms of adnormal on mushroom development.

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Jab1 as a Mediator of Nuclear Export and Cytoplasmic Degradation of p53

  • Lee, Eun-Woo;Oh, Wonkyung;Song, Jaewhan
    • Molecules and Cells
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    • v.22 no.2
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    • pp.133-140
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    • 2006
  • Jun activation domain-binding protein 1 (Jab1) is involved in various cellular mechanisms including development in Drosophila and mouse, cell cycle control and signal transduction pathways. Recent studies also determined that Jab1 functions as a nuclear exporter and inducer of cytoplasmic degradation for several proteins including p53, p27, capsid of West Nile virus, and Smad4/7 proteins. In particular, p53 is shown to bind to and to be exported into the cytoplasm by Jab1, which helps to maintain low levels of p53 under normal conditions. This review was undertaken in an effort to understand the biological significance of the homeostasis of p53 as maintained in the presence of Jab1. Based on our observations, we have provided potential mechanistic hypotheses for the nuclear export of p53 in coordination with Jab1 and the role of other factors in these processes.

A Method of Preparing Recombinant Fusion Antigen from Rotavirus and Norovirus

  • Oh, Ho-Kyung;Huh, Chul-Sung;Baek, Young-Jin;Yoo, Dong-Wan
    • Proceedings of the Korean Society for Food Science of Animal Resources Conference
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    • 2004.05a
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    • pp.353-356
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    • 2004
  • Rotavirus and Norovirus are major causative agents of acute diarrhea and gastroenteritis. In our study, Each viral RNA was isolated from the feces of patients for viral diarrhea in Korea, respectively. And cDNA library were constructed using RT-PCR. Also, cDNAs encoding VP8 derived from Rotavirus and Capsid protein derived from norovirus were subesequently cloned and expressed in Echerichia coli as a fusion antigen. Molecular weight of fusion antigen was approximately 60kDa. Also, substantial overexpression was accomplished. We yielded egg yolk lgY which is potentially useful in controlling of Rotavirus and Norovirus which are one of the most prevalent pathogenic viruses.

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Differentiation of Lymphocystis Disease Virus Genotype by Multiplex PCR

  • Kitamura Shin Ichi;Jung Sung-Ju;Oh Myung-Joo
    • Journal of Microbiology
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    • v.44 no.2
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    • pp.248-253
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    • 2006
  • Lymphocystis disease virus (LCDV) is the causative agent of lymphocystis disease. The viruses have been divided into three genotypes (genotype I for LCDV-1, II for Japanese flounder isolates, and III for rockfish isolates) on the basis of major capsid protein (MCP) gene sequences. In this study, we developed a multiplex PCR primer set in order to distinguish these genotypes. We also analyzed the MCP gene of a new LCDV isolate from the sea bass (SB98Yosu). Comparison of sequence identities between SB98Yosu and eight Japanese flounder isolates, revealed identity of more than 90.1 % at nucleotide level and 96.5% at deduced amino acid level, respectively. Phylogenetic analyses based on the MCP gene showed that SB98Yosu belongs to genotype II, along with Japanese flounder isolates. Multiplex PCR based on the MCP gene allowed us to identify these genotypes in a simple and rapid manner, even in a sample that contained two genotypes, in this case genotypes II and III.

First detection and genetic characterization of porcine parvovirus 7 from Korean domestic pig farms

  • Ouh, In-Ohk;Park, Seyeon;Lee, Ju-Yeon;Song, Jae Young;Cho, In-Soo;Kim, Hye-Ryung;Park, Choi-Kyu
    • Journal of Veterinary Science
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    • v.19 no.6
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    • pp.855-857
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    • 2018
  • Porcine parvovirus 7 (PPV7) was first detected in Korean pig farms in 2017. The detection rate of PPV7 DNA was 24.0% (30/125) in aborted pig fetuses and 74.9% (262/350) in finishing pigs, suggesting that PPV7 has circulated among Korean domestic pig farms. Phylogenetic analysis based on capsid protein amino acid sequences demonstrated that the nine isolated Korean strains (PPV-KA1-3 and PPV-KF1-6) were closely related to the previously reported USA and Chinese PPV7 strains. In addition, the Korean strains exhibit genetic diversity with both insertion and deletion mutations. This study contributes to the understanding of the molecular epidemiology of PPV7 in Korea.

Two putative novel serotypes of Tibet orbivirus isolated from Culicoides spp. in Yunnan, China

  • Ying-Liang Duan;Zhen Xing Yang;Yu Wen He; Le Li
    • Journal of Veterinary Science
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    • v.24 no.2
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    • pp.18.1-18.7
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    • 2023
  • Tibet orbivirus (TIBOV) was identified as a novel orbivirus in 2014. Antibodies against TIBOV were detected in cattle, Asian buffalo, and goats, while all the sequenced TIBOV strains were isolated from mosquitos and Culicoides. The known TIBOV strains have been classified into four putative serotypes. In this study, two TIBOV strains isolated from Culicoides spp. in Shizong County of Yunnan Province, China, were fully sequenced. The phylogenetic analysis of outer capsid protein 2 (VP2) indicated that these two viral strains belong to two novel putative serotypes of TIBOV. The updated putative serotypes may help in an investigation of the distribution and virulence of TIBOV.

Production of the Polyclonal Antibody That Recognizes the Mutant M Protein of Japanese Encephalitis Virus: Role of Its Charged Residues in Virus Production (일본뇌염바이러스의 Mutant M 단백질에 반응하는 다클론항체의 생산: 극성 아미노산 잔기의 바이러스 생산과정에서의 역할)

  • Kim, Jeong-Min;Yun, Sang-Im;Song, Byung-Hak;Kim, Jin-Kyoung;Lee, Young-Min
    • Korean Journal of Microbiology
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    • v.46 no.2
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    • pp.140-147
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    • 2010
  • Japanese encephalitis virus (JEV), a member of the mosquito-borne flaviviruses, causes epidemics of viral encephalitis in the Southeastern Asia. JEV is a small enveloped virus with a positive-sense RNA genome; the infectious virion consists of three structural proteins, namely capsid, membrane (M; a mature form of its prM precursor), and envelope proteins. Here, we investigated a role of the charged residues found at the N-terminus of the JEV M protein in virus production. Using an infectious JEV cDNA, we generated two mutant cDNAs, Mm1 and Mm2, by charged-to-alanine substitution for $E^9$ and $K^{15}K^{16}E^{17}$ residues of the M protein, respectively. By transfection of wild-type or each of the two mutant RNAs transcribed from the corresponding cDNAs, we found that Mm2, but not Mm1, had a ~3-log decrease in virus production, even though a comparable amount of all three structural proteins were produced in transfected cells. Interestingly, the prM protein expressed in Mm2 RNA-transfected cells was not recognized by the polyclonal antiserum raised against the N-terminal 44 amino acids of the wild type M protein, but reacted to the antiserum raised against the corresponding region of the mutant Mm2. Our results indicate that three charged residues ($K^{15}K^{16}E^{17}$) in JEV M protein play a role in virus production. Two polyclonal antisera specifically recognizing the wild-type or Mm2 version of the M protein would provide a useful reagent for the functional study of this protein in the virus life cycle.