• Title/Summary/Keyword: Capsid

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Comparison of lymphocystis disease virus proteins between marine and freshwater fish

  • Hossain, Mosharrof;Jung, Sung-Ju;Kim, Wi-Sik;Kim, Seok-Ryel;Oh, Myung-Joo
    • Journal of fish pathology
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    • v.22 no.2
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    • pp.173-177
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    • 2009
  • Lymphocystis disease virus (LCDV) was detected from olive flounder Paralichthys olivaceus, painted glass fish Chanda baculis, gourami Trichogaster leeri and rockfish Sebastes schlegeli, and proteins of the viruses were compared. The major capsid protein (MCP) gene-specific primer sets successfully amplified approximately 1300 bp nucleotides from the olive flounder and 600 bp nucleotides from painted glass fish, gourami and rockfish isolates, respectively. In western blotting analysis using anti-LCDV mouse polyclonal serum, major antigenic proteins had 21, 26, 45, 50, 80, 110 and 120 kDa in olive flounder, 26, 47 and 80 kDa in painted glass fish, 26, 46, 80 and 92 kDa in gourami, 26, 44, 49, 80 and 105 in rockfish, respectively. All the marine and freshwater isolates showed only common antigens of approximately 26 kDa and 80 kDa. These results suggest that antigenic protein profiles of LCDVs may vary depending upon fish species.

Characterization of rock bream (Oplegnathus fasciatus) fin cells and its susceptibility to different genotypes of megalocytiviruses

  • Jeong, Ye Jin;Kim, Young Chul;Min, Joon Gyu;Jeong, Min A;Kim, Kwang Il
    • Journal of fish pathology
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    • v.34 no.2
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    • pp.149-159
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    • 2021
  • Genus Megalocytivirus cause red sea bream iridoviral disease (RSIVD) and scale drop disease (SDD). Based on the phylogeny of the major capsid protein (MCP) and adenosine triphosphatase (ATPase) genes, megalocytiviruses except for SDD virus (SDDV) could be three different genotypes, red sea bream iridovirus (RSIV), infectious spleen and kidney necrosis (ISKNV), and turbot reddish body iridovirus (TRBIV). In this study, primary cells derived from the caudal fin of rock bream (Oplegnathus fasciatus) grew at 25℃ in Leibovitz's medium supplemented with 10% (v/v) fetal bovine serum and primocin (100 ㎍/mL). Rock bream fin (RBF) cells exhibited susceptibility to infections by different genotypes of megalocytiviruses (RSIV, ISKNV and TRBIV) with the appearance of cytopathic effects with an increase in the viral genome copy number. Furthermore, compared to grunt fin (GF) cells, even though 10 times lower number of RSIV genome copies were inoculated in RBF cells, viral genome copy number produced on RBF cells were 44 times higher than that of GF cells at 7 d post-inoculation. As the isolated RBF cells are sensitive to different genotypes of megalocytiviruses (RSIV, ISKNV and TRBIV), they can be used for future studies regarding in vitro viral infection and subsequent diagnosis.

Characterization of Prophages in Leuconostoc Derived from Kimchi and Genomic Analysis of the Induced Prophage in Leuconostoc lactis

  • Kim, Song-Hee;Park, Jong-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.32 no.3
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    • pp.333-340
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    • 2022
  • Leuconostoc has been used as a principal starter in natural kimchi fermentation, but limited research has been conducted on its phages. In this study, prophage distribution and characterization in kimchi-derived Leuconostoc strains were investigated, and phage induction was performed. Except for one strain, 16 Leuconostoc strains had at least one prophage region with questionable and incomplete regions, which comprised 0.5-6.0% of the bacterial genome. Based on major capsid protein analysis, ten intact prophages and an induced incomplete prophage of Leu. lactis CBA3626 belonged to the Siphoviridae family and were similar to Lc-Nu-like, sha1-like, phiMH1-like, and TPA_asm groups. Bacterial immunology genes, such as superinfection exclusion proteins and methylase, were found on several prophages. One prophage of Leu. lactis CBA3626 was induced using mitomycin C and was confirmed as belonging to the Siphoviridae family. Homology of the induced prophage with 21 reported prophages was not high (< 4%), and 47% identity was confirmed only with TPA_asm from Siphoviridae sp. isolate ct3pk4. Therefore, it is suggested that Leuconostoc from kimchi had diverse prophages with less than 6% genome proportion and some immunological genes. Interestingly, the induced prophage was very different from the reported prophages of other Leuconostoc species.

Pathogenicity and localization of the tobacco mosaic virus 4.8 kDa protein(oral)

  • Palukaitis, P.;Canto, T.;MacFarlane Scottish, S.A.
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.65.1-65
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    • 2003
  • In addition to the five well-characterized genes of Tobacco mosaic virus (TMV), this virus contains a sixth open reading frame (ORF6) that encodes a 4.8 kDa protein. TMV ORF6 overlaps the ORFs encoding the 30 kDa movement protein and the adjacent 17.5 kDa capsid protein. Although the 4.8 kDa protein could not be detected in vivo, alteration of the AUG codons of this ORF resulted in a mutant virus that attenuated the virulence of the mutated TMV in Nicotiana benthamiana, but not N. tabacum (tobacco). These sequence changes did not affect either the replication or movement of the mutated TMV. Expression of TMV ORF6 from the virus expression vector Potato virus X (PVX) intensified the virulence of this virus in N. benthmiana, but not tobacco, while expression of TMV ORF6 from the virus expression vector Tobacco rattle virus enhanced the pathogenicity observed in both N. benthamima and tobacco. Thus, the TMV ORF6 is a host- and virus-specific. virulence factor. However, two separate assays indicated that the TMV 4.8 kDa protein was not a suppression of RNA silencing. A fusion protein formed between the TMV 4.8 kDa protein and the green fluorescent protein was expressed from the PVX vector and localized to plasmodesmata. Possible roles of the 4.8 kDa protein in pathogenicity will be discussed

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Cholic Acid Attenuates ER Stress-Induced Cell Death in Coxsackievirus-B3 Infection

  • Han, Jae-Young;Jeong, Hae In;Park, Cheol-Woo;Yoon, Jisoo;Ko, Jaeyoung;Nam, Sang-Jip;Lim, Byung-Kwan
    • Journal of Microbiology and Biotechnology
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    • v.28 no.1
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    • pp.109-114
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    • 2018
  • Coxsackievirus Type B3 (CVB3) is an enterovirus that belongs to the Picornaviridae and causes various diseases such as myocarditis and hand-foot-mouth disease. However, an effective antiviral drug is still not developed. In this study, we looked for potential inhibitors of CVB3 replication by examining the survival of CVB3-infected HeLa cells. We detected an antiviral effect by cholic acid and identified it as a candidate inhibitor of CVB3 replication. Cholic acid circulates in the liver and intestines, and it helps the digestion and absorption of lipids in the small intestine. HeLa cells were cultured in 12-well plates and treated with cholic acid (1 and $10{\mu}g/ml$) and $10^6PFU/ml$ of CVB3. After 16 h post-infection, the cells were lysed and subjected to western blot analysis and RT-PCR. The production of the viral capsid protein VP1 was dramatically decreased, and translation initiation factor eIF4G1 cleavage was significantly inhibited by treatment with $10{\mu}g/ml$ cholic acid. Moreover, cholic acid inhibited ERK signaling in CVB3-infected HeLa cells. RT-PCR showed that the amounts of the CVB3 RNA genome and mRNA for the ER stress-related transcription factor ATF4 were significantly reduced. These results showed that cholic acid strongly reduced ER stress and CVB3 proliferation. This compound can be developed as a safe natural therapeutic agent for enterovirus infections.

Production of Specific Egg Yolk Antibodies in Chicken against Recombinant Fowl Adenovirus Fiber 2 Protein (재조합 가금 아데노바이러스 Fiber 2 단백질을 이용한 특이 난황 항체 생산)

  • Jung, Kyung Min;Lee, Seong;Kim, Jung Woo
    • Korean Journal of Poultry Science
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    • v.41 no.1
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    • pp.15-20
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    • 2014
  • Fowl adenovirus (FAV) is an important cause of several diseases, which result in considerable economic losses to the poultry farm. An outer capsid protein of FAV, fiber 2 is essential for virus growth, assembly or spread. This study was performed to produce about 22 kDa of recombinant fiber 2 protein and to immunize in laying hens to acquire the specific IgY antibody against the recombinant fiber 2. Laying hens were immunized with the recombinant fiber 2 intramuscularly in the breast muscle by injection 4 times at intervals of three weeks. At 12 weeks, serum- and egg yolk-antibody titers of hens against fiber 2 were increased up to 430,000 and 414,000, respectively. The recombinant fiber 2 could be recognized be the anti-His monoclonal antibody. Anti-fiber 2-IgY antibody could recognize the fiber 2 specifically in western blot analysis. These results suggested that the recombinant fiber 2 antigen could be used as an immunogen to elicit IgY antibody against fiber 2 and the anti-fiber 2-IgY could neutralize fowl adenovirus fiber 2 effectively.

Development of the Gene Therapy Vector for Targeting Ovarian Cancer Cells through ErbB Receptors (ErbB 수용체를 이용한 난소암세포 표적 유전자치료 벡터의 개발)

  • Joung, In-Sil;Bang, Seong-Ho
    • Korean Journal of Microbiology
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    • v.47 no.1
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    • pp.1-6
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    • 2011
  • Inefficiency of in vivo gene transfer using currently available vectors reflects a major hurdle in cancer gene therapy. Both viral and non-viral approaches have been described to improve gene transfer efficiency but suffer from a number of limitations. Here we tested an adenovirus carrying the small peptide ligand derived from heregulin${\beta}$ EGF-like domain onto fiber, the adenoviral capsid protein, to deliver transgene to ovarian cancer cells which overexpress ErbB, the cognate receptors for heregulin. The attachement of 53 amino acids to fiber didn't affect on the fiber's trimer structure that is critical for the viral entry to cells. The fiber-modified adenovirus can mediate entry and expression of a ${\beta}$-galactosidase into cancer cells in an increased efficiency compared the unmodified adenovirus. Particularly, the gene transfer efficiency was improved up to 5 times in OVCAR3 cells, an ovarian cancer cell line. Such transduction systems hold promise for delivering genes to ErbB receptor overexpressing cancer cells, and could be used for future cancer gene therapy.

White Sport Syndrome Virus Disease of Shirmp and Diagnostic Methods

  • Zhan, Wen-Bin
    • Journal of Aquaculture
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    • v.15 no.1
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    • pp.7-13
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    • 2002
  • Since 1993, the White Spot Syndrome Virus (WSSV) disease occurred in China among cultured shrimps resulting in mass mortality. Epizootiological surveys undertaken during the outbreak period of 1993-1994 indicated that all stages of Penaeus chinensis, P. japonicus and P. monodon were infected. Consequent to the transport of contaminated shrimp seedlings and seawater, the disease spread all over the farms of China. The disease was more rapidly transmitted at temperatures above $25^{\circ}C$. Challenge experiments showed the causative agent was highly virulent. White spots appeared on the carapace of both span-taneous and experimentally infected shrimps. Moribund shrimps contained turbid hemolymph, hypertrophied Iymphoid organ and a necrotic mid-gut gland. Electron microscopy showed the presence of viral particles in the gills, stomach, lymphoid organ, and epidermal tissue of the infected shrimp. The visions were slightly ovoid with an envelope and averaged 350 $\times$ 150 nm; nucleocapsids measured 375 $\times$ 157 nm. With discontinuous sucrose gradient of 35, 50 and 60% (w/v), the virus was separated from hemolymph of the infected shrimp. The estimated molecular weight of genomic DNA was 237 Kb with EcoR I, 247 Kb with Hind III and 241kb with Pst I. A total of 9 hybridoma colones secreting monoclonal antibodies (MAbs) were produced from mouse myeloma and spleen cells immunized with WSSV. The immunofluorescence assay of gill tissue showed that the MAbs reacted with diseased but not with healthy shrimp. The MAbs belonged to IgGl, IgG2b subclass and IgM class, all with kappa light Immune-electron-microscopy with colloidal gold marker showed the presence of 5 MAbs epitopes on the envelope and one on the capsid of the virus. Baculoviral mid-gut gland necrosis showed the specificity of the MAbs produced. For diagnosis 5 different methods were selected. Using Kimura primers for PCR, or MAbs for immunoblot, ELISA or FAT method, in situ hybridization was carried out to show the gene. All these methods detected WSSV in the organ samples of the diseased shrimp but not in healthy one.

Clinical features of Epstein-Barr virus-associated infectious mononucleosis in hospitalized Korean children

  • Son, Keun-Hyung;Shin, Mee-Yong
    • Clinical and Experimental Pediatrics
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    • v.54 no.10
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    • pp.409-413
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    • 2011
  • Purpose: Few studies have been conducted on the recent status of infectious mononucleosis (IM) in Korean children. The aim of this study was to evaluate the recent trend in the clinical manifestations of Epstein-Barr virus (EBV)-associated IM as well as the clinical differences according to age. Methods: A retrospective study was performed on 81 children hospitalized with EBV-associated IM who fulfilled the serological criteria for the diagnosis of EBV infection (viral capsid antigen immunoglobulin M positive). The patients were divided into 3 age groups: <5 years, 5 to 9 years, and ${\geq}10$ years. We evaluated the recent trend in clinical manifestations and the differences in clinical and laboratory findings among the 3 age groups. Results: Thirty (37%) children were under 5 years of age, 38 (46.9%) were 5 to 9 years of age, and 13 (16%) were 10 years of age or older. The differences in the symptoms and signs among the 3 age groups were not statistically significant, except for headache. The mean duration of fever was 7.7 days (range, 0 to 18 days). A comparison of liver enzyme elevation among the age groups showed an association with advancing age (26.6%, 63.1%, and 76.9%, respectively, P=0.04) Conclusion: This study showed that EBV-associated IM in Korean children continues to occur mostly in children under 10 years of age. In children with EBV-associated IM, the incidence of headache and liver enzyme elevation, the duration of fever, and the proportion of females to males were all positively associated with advancing age.

Effect of Hydrogel on Survial of Serratia plymuthica A21-4 in Soils and Plant Disease Suppression

  • Shen, Shun-Shan;Kim, Won-Il;Park, Chang-Seuk
    • The Plant Pathology Journal
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    • v.22 no.4
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    • pp.364-368
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    • 2006
  • Survival of biocontrol agents and their effective colonization of rhizhosphere are the essential components for successful disease suppression. The effects of hydrogel supplement on bacterial survival and disease control were evaluated in pot and in the field. Addition of 2% hydrogel material to potting soil resulted in significant enhancement of colonization of biocontrol agent Serratia plymuthica A21-4 both in soil and rhizosphere of pepper plants. Rhizosphere colonization of S. plymuthica A21-4 retrieved from 40 days old pepper seedlings indicated 100 times higher bacterial population in hydrogel treated soil than in ordinary pot soil. The pepper plants sown in hydrogelated potting soil showed higher seed germination rate and the better growth of pepper plant than those in ordinary commercial pot soil. Although the suppression of Phytophthora capsid density in the potting soil by treatment of biocontrol agent A21-4 was not significantly different between in hydrogelated soil and ordinary potting soil, the suppression of Phytophthora blight between two treatments was significantly different. A21-4 treatment in hydrogelated potting soil was completely disease-free while same treatment in ordinary potting soil revealed 36% disease incidence. Our field study under natural disease occurrence also showed significantly less disease incidence(12.3%) in the A21-4 treatment in the hydrogelated soil compared to other treatments. Yield promotion of pepper by the A21-4 treatment in the hydrogelated potting soil was also recognized. Our results indicated that hydrogel amendment with biocontrol agent in pot soil would be a good alternative to protect pepper seedlings and increase plant yield.