• 한국어병학회지
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• 제22권2호
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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.

• 한국어병학회지
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• 제34권2호
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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.

• Journal of Microbiology and Biotechnology
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• 제32권3호
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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.

• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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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

• Journal of Microbiology and Biotechnology
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• 제28권1호
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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.

• 한국가금학회지
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• 제41권1호
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• pp.15-20
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• 2014

본 연구는 가금에서 호흡기 질환, 심낭수종, 봉입체 간염, 산란저하증 등을 유발하는 가금 아데노바이러스의 fiber 2 유전자에 대한 특이 난황 항체를 개발하고자 실험을 실시하였다. Fiber 2 유전자를 클로닝한 뒤, 대장균 발현 시스템을 이용하여 약 22 kDa의 재조합 fiber 2 단백질을 생산하였다. 이를 산란계에 3주 간격으로 총 4회 면역하여 혈청 및 난황 내 항체가를 측정한 결과, 면역 후 12주경에 항체가가 최고치에 달하였으며, 산란계로부터 획득한 난황 항체를 이용한 Western blot analysis 결과, 가금 아데노바이러스 내 fiber 2 단백과 특이적으로 반응한다는 것을 규명하였다. 결론적으로, 가금 아데노바이러스 fiber 2에 특이적인 난황 항체 생산에 성공하였으며, 이러한 특이적인 난황 항체는 가금 아데노바이러스로 인한 질병의 예방 및 치료에 활용 가능할 것으로 사료된다.

• 미생물학회지
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• 제47권1호
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• pp.1-6
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• 2011

암의 유전자치료에서 암세포로의 선택적 유전자전달 매체의 부족은 치료효과의 감소를 야기하는 문제이다. 본 연구에서는 난소암 유전자치료의 효율을 높이기 위한 목적으로 난소암세포로 선택적인 유전자전달을 하도록 개량된 아데노바이러스 벡터를 제조하고, 그 효율성을 난소암세포주를 이용하여 조사하였다. 난소암세포에 과다발현하는 분자인 ErbB receptor를 표적하도록 아데노바이러스 외피단백질 fiber에 ErbB receptor에 대한 ligand인 heregulin으로부터 유래한 펩티드를 부착하였다. 53개의 아미노산으로 구성된 외부 펩티드를 fiber에 부착하였을 때 바이러스 감염에 중요한 기능을 하는 fiber 삼량체 구조 형성에 영향을 미치지 않았다. Fiber를 조작한 개량 아데노바이러스는 야생형 fiber를 가진 1세대 아데노바이러스 벡터에 비해 선택적으로 난소암으로 유전자를 전달하는 비율이 증가하였다. 특히 항암제에 저항성을 가진 난소암세포주 OVCAR3에서 유전자전달 효율이 약 5배 증가되었다. 따라서 난소암의 유전자치료에서 개량된 아데노바이러스로 치료 유전자를 전달하면 치료의 효율성을 향상시킬 수 있을 것이다.

• 한국양식학회지
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• 제15권1호
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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 and Experimental Pediatrics
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• 제54권10호
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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.

• The Plant Pathology Journal
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• 제22권4호
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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.