• Title/Summary/Keyword: capsid

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First Isolation and Molecular Characterization of Bacteriophages Infecting Acidovorax citrulli, the Causal Agent of Bacterial Fruit Blotch

  • Rahimi-Midani, Aryan;Lee, Yong Seok;Kang, Se-Won;Kim, Mi-Kyeong;Choi, Tae-Jin
    • The Plant Pathology Journal
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    • v.34 no.1
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    • pp.59-64
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    • 2018
  • Bacteriophages of Acidovorax citrulli, the causal agent of bacterial fruit blotch, were isolated from 39 watermelon, pumpkin, and cucumber leaf samples collected from various regions of Korea and tested against 18 A. citrulli strains. Among the six phages isolated, ACP17 forms the largest plaque, and exhibits the morphology of phages in the Myoviridae family with a head diameter of $100{\pm}5nm$ and tail length of $150{\pm}5nm$. ACP17 has eclipse and latent periods of $25{\pm}5min$ and $50{\pm}5min$, respectively, and a burst size of 120. The genome of ACP17 is 156,281 base pairs with a G + C content of 58.7%, 263 open reading frames, and 4 transfer RNA genes. Blast search and phylogenetic analysis of the major capsid protein showed that ACP17 has limited homology to two Stentrophomonas phages, suggesting that ACP17 is a new type of Myoviridae isolated from A. citrulli.

Cytologic Findings of Infectious Mononucleosis Lymphadenitis - A Report of Four Cases - (전염성 단핵구중 림프절염의 세침흡인 세포학적 소견 - 4예 보고 -)

  • Sohn, Jin-Hee;Jung, Eun-Ha;Park, Hye-Rim
    • The Korean Journal of Cytopathology
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    • v.9 no.2
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    • pp.227-231
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    • 1998
  • Infectious mononucleosis(IM) Is an acute self-limiting lymphoproliferative disorder associated with infection by the Epstein-Barr Virus(EBV), with the characteristic triad of fever, sore throat, and cervical or generalized lymphadenopathy. And also there are atypical lymphocytes in the peripheral blood. Cytological findings of IM lymphadenitis are characterized by a florid immunoblastic and atypical lymphoid cell proliferation. However, the small number of cases were studied by fineneedle aspiration cytology(FNAC) even though there was a complexity of lymph node pathology. It is important to recognize the reactive pattern of IM that would initiate EBV study and to avoid unnecessary biopsy We studied findings of lymph node FNAC from 4 patients with EBV infection confirmed by EBV-specific serologic studios. All of the cases were positive for viral capsid antigen(VCA) and one case was positive for anti-EBV nuclear antigen(EBNA). Cytologically, all of the cases exhibited high cellularity and atypia with great numbers of large immunoblastic lymphocytes.

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ORI2 is a Strong Inhibitor of Coxsackievirus B4 Replication (오리방풀로부터 분리된 ORI2의 췌장염 유발 콕사키바이러스B4 증식억제)

  • Lim, Byung-Kwan;Jo, Soyeon;Kim, Jin Hee
    • Korean Journal of Pharmacognosy
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    • v.45 no.4
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    • pp.282-287
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    • 2014
  • The ORI2 (3-[3,4-dihydroxyphenyl]acrylic acid 1-[3,4-dihydroxyphenyl]-2-methoxycarbonylethyl ester) was purified from the extract of Isodon excisus. We confirmed the antiviral effect of ORI2 in a coxsackievirus-induced pancreatitis model. Coxsackievirus B4 (CVB4) is a common cause of pancreatitis and may be reason of the type-1 diabetes. Anti-enteroviral compounds were screened by HeLa cell survival assay. Purified natural compounds were added to HeLa cells cultured 96-well plates after $10^4PFU/ml$ CVB4 pre-incubation for 30 min. ORI2 significantly improved HeLa cell survival in a dose-dependent manner. In addition, ORI2 (1 mM) treatment was dramatically decreased virus protease 2A induced eIF4G-I cleavage and viral VP1 capsid protein production. HeLa cell virus titers and viral RNA replication were significantly decreased in ORI2-treatment in a dose dependent manner (1 mM~0.001 mM). These results demonstrate that ORI2 has a strong antiviral effect. It was significantly decreased virus replication. ORI2 may be developed as a potential therapeutic agent for CVB4.

Analysis of antigenic sites on the VP4 of porcine rotavirus, Gottfried strain (돼지 로타바이러스(Gottfried 주)의 VP4 항원구조분석)

  • Song, Yun-kyung;Kim, Won-yong;Kang, Shien-young
    • Korean Journal of Veterinary Research
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    • v.41 no.3
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    • pp.343-350
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    • 2001
  • The neutralization epitopes of the outer capsid protein VP4 of a porcine rotavirus, Gottfried strain, were studied using neutralizing monocolonal antibodies(N-MAbs). Eight N-MAbs which are specific for the VP4 of Gottfried strain were used for analyzing the antigenic sites of VP4. Three different approaches were used for this analysis; i)testing the serological reactivity of each N-MAb against different G and P types of human and animal rotavirusese ii) analyzing N-MAb-resistant viral escape mutants and iii) performing nucleotide sequence analysis of the VP4 gene of each N-MAb-resistant viral escape mutant. From experimental results, at least four antigenic sites(I, II, III, and IV) were identified. Antigenic site I recognized by N-MAbs 24B9, 23G10, and 26A2 was separated from antigenic site II recognized by N-MAbs 30H5, 32B3, and 29B3. However, these antigenic sites were overlapped with antigenic site III recognized by N-MAb 21A1. The other antigenic site IV recognized by N-MAb 16D2 was separated from antigenic sites I, II, and III.

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Studies on the RK-temperate phage of bacillus cereus (Bacollis cereis의 RK-용원파아지에 관한 연구)

  • 이태우
    • Korean Journal of Microbiology
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    • v.23 no.2
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    • pp.129-137
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    • 1985
  • The RK-temperate phage which infected with Bacillus cereus was isolated and the characters were investigated. The induction of RK-temperate phage from host bacterium attained by ultraviolet light irradiation (15W, 30cm, 30-120sec) and mitomycin C treatment (0.2-2 ug/ml). The host range of RK-temperate phage was not revealed with lysogenic and related strains of B. cereus. But B. cereus(PS) 352 which obtained by N-nitrosoguanidine treatment (1,000{$\mu}g/ml)$ to phage infected with host bacteria was sensitive bacteria of RK-temperate phage. RK-temperate phage was stabilized at the condition of nutrient broth (pH 7-8), Tris-buffer (pH 7-8) and ammonium buffer (pH 8-9) and Sorensen's phosphate buffer (pH 6-7), but unstabilized at other salt solutions and pH range. Also, thermostability was to $45^{\circ}C$ but unstabilized at above $50^{\circ}C$. At RK-temperate phage, the measurment values of head, neck, mid tail and end tail were 59nm, $9{\times}16nm,\;10{\times}189nm,\;and\;10{\times}14nm$ respectively. The morphology of head was regular polyhedron, and the end tail was coneate form. On the one hand, the number of capsid protein layer of tail were consist of 4, 35, and 1 at neck, mid tail, and end tail, respectively. RK-temperate phage was identified with DNA phage and G+C contents were 38.63. The latent time of RK-temperate phage was 30 minutes and the burst size was 70-80. And the host bacteria was lysed in case of multi-infection, above moi 1.

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RT-PCR Detection of Five Quarantine Plant RNA Viruses Belonging to Potyand Tospoviruses

  • Lee, Jong-Seung;Cho, Won-Kyong;Choi, Hong-Soo;Kim, Kook-Hyung
    • The Plant Pathology Journal
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    • v.27 no.3
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    • pp.291-296
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    • 2011
  • In order to detect quarantine plant viruses, we developed reverse transcription-polymerase chain reaction (RT-PCR) primer pairs for five single-stranded (ss) plant RNA viruses that are not currently reported in Korea but could be potential harmful plant viral pathogens. Three viruses such as Chilli veinal mottle virus (ChiVMV), Colombian datura virus (CDV), and Tobacco etch virus (TEV) belong to the genus Potyvirus while Chrysanthemum stem necrosis virus (CSNV) and Iris yellow spot virus (IYSV) are members of the genus Tospovirus. To design RT-PCR primers, we used reported gene sequences corresponding to the capsid protein and polyprotein for ChiVMV, CDV, and TEV while using nucleocapsid protein regions for CSNV and IYSV. At least two different primer pairs were designed for each virus. Fifteen out of 16 primer pairs were successfully applied in detection of individual quarantine virus with high specificity and efficiency. Taken together, this study provides a rapid and useful protocol for detection of five quarantine viruses.

Enhancement of Biocontrol Activity of Antagonistic Chryseobacterium Strain KJ1R5 by Adding Carbon Sources against Phytophthora capsici

  • Kim, Yu-Seok;Jang, Bo-Ra;Chung, Ill-Min;Sang, Mee-Kyung;Ku, Han-Mo;Kim, Ki-Deok;Chun, Se-Chul
    • The Plant Pathology Journal
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    • v.24 no.2
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    • pp.164-170
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    • 2008
  • Carbon utilization by Chryseobacterium strain KJ1R5 was studied to enhance its biocontrol activity against Phytophthora capsid. Chryseobacterium strain KJ1R5 has previously been shown to control Phytophthora blight of pepper (Capsicum annuum L.). Strain KJ1R5 could utilize carbon sources such as L-arabinose, D-cellobiose, ${\beta}-lactose$ and D-galactose well. P. capsici could utilize D-glucose well, showing the absorbencies ranged from 0.577 to 0.767 at 600nm. When 2% L-arabinose, which could only be utilized by the bio-control strain KJ1R5, was amended into the bacterial suspension, the efficacy of biological control increased. Among the amendments of various carbon sources into bacterial suspension, L-arabinose and D-(+)-glucose significantly enhanced biological control activity, resulting in a reduction of disease incidence to 6.9%, compared to 21.9% for the strain KJ1R5 alone and 81.3% for P. capsici inoculation alone, indicating that amendment with specific carbon sources could increase the biological control activity.

Kaempferol Inhibits Enterovirus Proliferation through MAPK Signal Regulation (Kaempferol의 MAPK 신호 조절을 통한 심근염 유발 엔테로바이러스 증식 억제)

  • Jang, Jin-Hwa;Jeong, Hae-In;Lim, Byung-Kwan;Nam, Sang-Jip
    • Korean Journal of Pharmacognosy
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    • v.48 no.3
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    • pp.173-178
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    • 2017
  • We investigated the efficacy of single compound of plant extract in coxsackievirus B3 (CVB3) infection. CVB3 is a main cause of Hand-foot-mouth diseases (HFMD) and viral myocarditis in children and adult. Several single compounds of plant extract were purified by HPLC and tested as antiviral drug candidate. Among them, kaempferol was selected to effective anti-enterovirus compound by HeLa cells survival assay. CVB3 infected HeLa cells were treated with kaempferol ($100{\mu}g/ml-100ng/ml$) and their antiviral effect was confirmed. After 16 hours of treatment, HeLa cells were lysed and proteins were extracted for western blot analysis. CVB3 viral capsid protein VP1 production and transcription factor eIF4G-1 cleavage was significantly decreased in $100{\mu}g/ml$ kaempferol treatment. Virus replication was observed by virus RNA amplification. Kaempferol strongly reduced virus positive and negative strand RNA amplification. Moreover, MAPK signal induced by CVB3 infection, pERK and pmTOR, kaempferol treatment significantly inhibited the activity. Plant extract single compound, kaempferol, is a strong candidate to be developed non-toxic anti-enterovirus treatment agent.

Identification of antigenic proteins of lymphocystis disease virus (LCDV) by MALDI-TOF mass spectrometry

  • Chung, Chang-Kyun;Kim, Byung-Gwan;Jung, Myung-Hwa;Jung, Sung-Ju
    • Journal of fish pathology
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    • v.28 no.3
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    • pp.133-143
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    • 2015
  • The antigenic proteins of Lymphocystis disease virus (LCDV) from tumors of olive flounder, Paralichthys olivaceus, are described following characterization by mass spectrometry. In SDS-PAGE, predominant protein bands were observed at 114, 88, 70, 54, 52, 47, 42 and 24 kDa. Western blot analysis showed that antisera reacted strongly at molecular weights of 114, 67 and 54 kDa, and reacted weakly at molecular weights of 74, 70, 36, 24 and 22 kDa. In the identification of LCDV antigenic proteins by matrix-assisted laser desorption ionization (MALDI) TOF mass spectrometry, 10 of 14 excised bands consisted mostly of proteins with amino acid sequences that matched LCDV-C (lymphocystis disease virus isolate China) ORFs. Strong antigens with molecular weights of 114, 67 and 54 kDa were identified as LDVICp236 (chromosome segregation ATPase), LDVICp033 (membrane bound metallopeptidase) and LDVICp157 (hypothetical protein), respectively. Minor antigens with molecular weights of 70, 36, 24 and 22 kDa proteins were identified as LDVICp160 (acetyl-coA hydrolase), LDVICp213 (hypothetical protein), LDVICp039 (hypothetical protein) and LDVICp213 (hypothetical protein). However, the major capsid protein (LDVICp043) did not react with the polyclonal antibody.

A Newly Isolated Bacteriophage, PBES 02, Infecting Cronobacter sakazakii

  • Lee, Hyung Ju;Kim, Wan Il;Kwon, Young Chan;Cha, Kyung Eun;Kim, Minjin;Myung, Heejoon
    • Journal of Microbiology and Biotechnology
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    • v.26 no.9
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    • pp.1629-1635
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    • 2016
  • A novel bacteriophage, PBES 02, infecting Cronobacter sakazakii was isolated and characterized. It has a spherical head of 90 nm in diameter and a tail of 130 nm in length, and belongs to Myoviridae as observed under a transmission electron microscope. The major virion protein appears to be 38 kilodaltons (kDa) in size. The latent period of PBES 02 is 30 min and the burst size is 250. Infectivity of the phage remained intact after exposure to temperatures ranging from 4℃ to 55℃ for 1 h. It was also stable after exposure to pHs ranging from 6 to 10 for 1 h. The phage effectively removed contaminating Cronobacter sakazakii from broth infant formula. PBES 02 has a double-stranded DNA genome of 149,732 bases. Its GC ratio is 50.7%. Sequence analysis revealed that PBES 02 has 299 open reading frames (ORFs) and 14 tRNA genes. Thirty-nine ORFs were annotated, including 24 related to replication and regulation functions, 10 related to structural proteins, and 5 related to DNA packaging. The genome of PBES 02 is closely related to that of two other C. sakazakii phages, CR3 and CR8. Comparison of DNA sequences of genes encoding the major capsid protein revealed a wide geographical distribution of related phages over Asia, Europe, and America.