• 제목/요약/키워드: dsRNA virus

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Characterization of Cucumber mosaic virus Subgroup II Isolated from Paprika (Capsicum annuum var, grossum) in Korea

  • Choi, Gug-Seoun;Kim, Jae-Hyun;Choi, Jang-Kyung
    • The Plant Pathology Journal
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    • 제18권1호
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    • pp.6-11
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    • 2002
  • An isolate of Cucumber mosaic virus (CMV), PaFMl-CMV causing malformation on the fruit of paprika (Capsicum annuum var, grossum) was characterized based on biological reactions, serological relationships, and partial nucleotide sequence analyses. PaFMl-CMV was distinguishable from other isolates of CMYI Mf-(subgroup I) and LS-CMV (subgroup II), in terms of its reactions to some host plants. Polyclonal antibody against PaFMl-CMV showed homologous antigenic relationship with LS-CMV, however, the antibody formed a spur between PaFMl- and Mf-CMV, In the comparison of molecular size of dsRNAs of PaFMl-CMV with Mf- and LS-CMV, PaFMl-CMV had a slightly smaller RNAl and larger RNA2, RNA3, and RNA4. When the CDNA product of PaFMl-CMV coat protein (CP) gene was digested with some restriction enzymes, the fragment pattern was identical with that of LS-CMV The nucleotide and amino acid sequences of PaFMl-CMV CP gene were 99.5% and 98.6% identical with LS-CMV respectively. The data indicate that PaFMl-CMV belongs to subgroup II of CMV, which is the first report in Korea.

Finding and Characterization of Viral Nonstructural Small Protein in Prospect Hill Virus Infected Cell

  • 남기연;정동훈;최재원;이윤성;이평우
    • 대한바이러스학회지
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    • 제29권4호
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    • pp.221-233
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    • 1999
  • Prospect Hill Virus (PHV) is the well known serotype of hantavirus, a newly established genus in family Bunyaviridae. Extensive studies have upheld the original view of PHV genetics with three genes such as nucleocapsid (N) protein, envelope proteins (G1, G2) and RNA dependent RNA polymerase. In this study, we report the existence of additional gene that is encoded in an overlapping reading frame of the N protein gene within S genome segment of PHV. This gene is expected to encode a nonstructural small (NSs) protein and it seems to be only found in PHV infected cell. The presence and synthesis of NSs protein could be demonstrated in the cell infected with PHV using anti-peptide sera specific to the predicted amino acid sequence deduced from the second open reading frame. Ribosomal synthesis of this protein appears to occur at AUG codon at the 83rd base of S genome segment, downstream of N protein initiation codon. This protein is small in size (10.4 KDa) and highly basic in nature. The expression strategy of NSs protein appears that a signal mRNA is used to translate both N and NSs protein in PHV infected cell. 10 KDa protein in virus infected cell lysates can bind to mimic dsRNA. This fact strongly suggests that NSs protein may be involved in virus replication on late phase of viral life cycle.

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옥수수에 발생한 벼검은줄오갈병 바이러스 분절게놈 S8 및 S10 전 염기서열 분석 (Sequence Analysis of Segments 8 and 10 of Rice black-streaked dwarf virus from Maize Plants)

  • 이봉춘;조상윤;윤영남;강인정;곽도연;신동범;강항원
    • 식물병연구
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    • 제18권4호
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    • pp.387-390
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    • 2012
  • 2011년에 고창 옥수수 재배지에서 RBSDV에 의한 벼 검은줄오갈병이 발생하였다. 병징은 키가 작아지고 잎에 줄무늬가 형성되면서 심하면 거의 수확이 되지 않는다. 이병주로부터 직접 dsRNA를 추출하여 RT-PCR에 의해 감염을 확인하였다. S8 및 S10 분절의 전 게놈에 특이적인 primer를 이용하여 유전자를 증폭하였다. 이것을 크로닝한 후 전 염기서열을 결정하였다. 염기서열 분석결과 S8 및 S10의 전염기서열은 각각 1,936 nt 및 1,801 nt로 구성되었다. 이는 벼에서 분리된 RBSDV와 같은 크기로 확인되었다. 벼에서 분리된 RBSDV와 상동성 분석결과 S8은 94.9-99.6%, S10은 94.1-98.4%로 나타났다. Phylogenetic tree 분석결과 옥수수에서 분리한 RBSDV S8 및 S10 모두 벼에서 분리된 RBSDV와 같은 그룹으로 분류되었다.

Differential Protein Expressions in Virus-Infected and Uninfected Trichomonas vaginalis

  • Ding, He;Gong, Pengtao;Yang, Ju;Li, Jianhua;Li, He;Zhang, Guocai;Zhang, Xichen
    • Parasites, Hosts and Diseases
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    • 제55권2호
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    • pp.121-128
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    • 2017
  • Protozoan viruses may influence the function and pathogenicity of the protozoa. Trichomonas vaginalis is a parasitic protozoan that could contain a double stranded RNA (dsRNA) virus, T. vaginalis virus (TVV). However, there are few reports on the properties of the virus. To further determine variations in protein expression of T. vaginalis, we detected 2 strains of T. vaginalis; the virus-infected ($V^+$) and uninfected ($V^-$) isolates to examine differentially expressed proteins upon TVV infection. Using a stable isotope N-terminal labeling strategy (iTRAQ) on soluble fractions to analyze proteomes, we identified 293 proteins, of which 50 were altered in $V^+$ compared with $V^-$ isolates. The results showed that the expression of 29 proteins was increased, and 21 proteins decreased in $V^+$ isolates. These differentially expressed proteins can be classified into 4 categories: ribosomal proteins, metabolic enzymes, heat shock proteins, and putative uncharacterized proteins. Quantitative PCR was used to detect 4 metabolic processes proteins: glycogen phosphorylase, malate dehydrogenase, triosephosphate isomerase, and glucose-6-phosphate isomerase, which were differentially expressed in $V^+$ and $V^-$ isolates. Our findings suggest that mRNA levels of these genes were consistent with protein expression levels. This study was the first which analyzed protein expression variations upon TVV infection. These observations will provide a basis for future studies concerning the possible roles of these proteins in host-parasite interactions.

열무에서 분리한 오이모자이크바이러스 분리주의 특성 (Characterization of an Isolate of Cucumber mosaic virus from Raphanus sativus L.)

  • 이선주;홍진성;최장경;김은지;이긍표
    • 식물병연구
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    • 제17권2호
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    • pp.211-215
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    • 2011
  • 모자이크증상을 나타내는 열무로부터 Cucumber mosaic virus의 한 계통을 분리하고 특성을 조사하였다. Vigna unguiculata, Chenopodium amaranticolor and Gomphrena globosa에서 분리 바이러스의 기주반응과 dsRNA 분석, RT-PCR 검정, PCR-RFLP, 혈청학적 분석을 통하여 CMV의 한 계통임을 확인하였다. 이 CMV 계통을 다양한 지표식물에 접종하였을 때, Nicotiana benthamiana와 N. glutinosa, N. tabacum, 고추, 오이 그리고 멜론에서는 전형적인 강한 모자이크 병징이 나타났으나, 열무, 배추, 적갓은 매우 약한 병징을 나타내는 특징을 보였다. 새롭게 분리된 CMV 계통은 가지과, 박과 및 배추과 등 광범위한 작물에 감염성을 나타내었으며, 배추과에서의 감염성 차이를 근거로 Gn-CMV로 명명하였다. Double-stranded (ds) RNA를 분리한 분석에서 Gn-CMV는 기 보고된 CMV 계통과 마찬가지로 3.3, 3.0, 그리고 2.2 kb의 독립된 게놈으로 구성되어 있었으며, SDS-PAGE와 Western blotting 분석으로 통하여 28 kDa의 외피단백질을 확인할 수 있었다. RT-PCR로 얻어진 증폭산물을 Cac8I, ClaI and MspI을 이용한 PCR-RFLP를 통하여 CMV subgroup I 임을 확인할 수 있었다.

A Novel Strain of Cucumber mosaic virus Isolated from Lilium longiflorum

  • Jung, Hye-Jin;Ueda, Shigenori;Ryu, Ki-Hyun;Lee, Sang-Yong;Choi, Jang-Kyung
    • The Plant Pathology Journal
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    • 제16권6호
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    • pp.306-311
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    • 2000
  • A new strain of Cucumber mosaic virus (CMV) from easter lily (Lilium longiflorum), Ly2-CMV, was identified and compared to the well-characterized Mf-CMV (subgroupⅠ) and LS-CMV (subgroupⅡ) by host reaction in several indicator plants, dsRNA analysis, serological property, RT-PCR analysis, restriction enzyme profile of the PCR products and nucleotide sequence of coat protein (CP) gene. Remarkable differences in symptoms of Ly2-CMV were found between Mf-CMV or LS-CMV in tobacco plants and Datura stramoinium. Ly2-CMV induced small necrotic ringspots on the inoculated leaves of Nicotiana tabacum cvs. Xanthi nc and Burley 21 and D. stramonium, and failed to infect these species systemically. Of the indicator plants tested, N. benthamiana only reacted with systemic infection by inoculation of Lr2-CMV. In experiments of dsRNA analysis, serology and RT-PCR of CP gene, Ly2-CMV was come within subgroupⅠ CMV. However, restriction enzyme analysis of the PCR products using MspⅠ showed that Ly2-CMV was distinct to Mf-CMV. The CP gene of Ly2-CMV contains 657 nucleotides, and the nucleotide sequence is similar to that of Mf-CMV. There is also a high degree of conservation between their putative gene products in Ly2-CMV and Mf-CMV, with five amino acid changes in the 218 amino acids of the CPs.

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PT-PCR 법에 의한 Infectious Pancreatic Necrosis Virus의 조기진단 (Rapid and Sensitive Detection of Infectious Pancreatic Necrosis Virus (IPNV) by Revers Transcription-Polymerase Chain Reaction (RT-PCR))

  • 강호성;공희정;구현나;박정우;손상규;박명애;김한도
    • 한국양식학회지
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    • 제10권2호
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    • pp.171-178
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    • 1997
  • Infectious pancreatic necrosis virus (IPNV)는 치어에 감염되어 치명적인 질병을 유발하는, 양식산업에 있어 중요한 어류 병원체이다. 본 연구에서는 IPNV를 신속, 정확하게 진단하는 방법을 개발하고자 IPNV의 항원성 단백질인 VP2 유전자 부분에서 선택한 primers를 이용하여 역전사-중합효소연쇄반응법(Reverse Transcription-Polymerase Chain Reaction, RT-PCR)을 실시하였다. RT-PCR 증폭법으로 순수분리도니 IPNV dsRNA 40 ng 정도의 적은 양도 확인 할 수 있었으며, IHNV와 같은 다른 어류 병원체의 게놈을 RT-PCR templates로 사용하였을 경우는 어떠한 PCR 산물도 검출되지 않는 특이성을 보였다. 특히 유전자의 분리없이 조직 그 자체를 대상으로 RT-PCR을 행하는 in situ RT-PCR 방법으로 IPNV가 감염된 넙치 (Paralichthy olivaceus) 치어의 조직에서 IPNV 감염을 신속하게 확인할 수 있었다. 따라서 RT-PCR 및 in situ RT-PCR 방법은 IPNV를 신속, 정확하게 진단하는데 활용될 수 있을 것으로 생각된다.

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Biochemical Characteristics of a Killer Toxin Produced by Ustilago maydis Virus SH14 Isolated in Korea

  • Ha, Eun-Soo;Yie, Se-Won;Choi, Hyoung-Tae
    • Journal of Microbiology
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    • 제35권4호
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    • pp.323-326
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    • 1997
  • Toxin protein from Ustilago maydis virus SH14 isolated in Korea was purified using ethanol precipitation, cation exchange, gel filtration and anion exchange chromatography. The molecular weight of the purified protein was estimated to be 8.3 kDa by SDS-PAGE analysis. The Nterminal sequence of the protein is L-G-I-N-C(K)-R-G-S-S-Q--C(K)-G-L-S-G which is highly homologous with that of P4 toxin, but the amino acid composition and electrophoretic mobility in a native PAGE of the toxin protein were totally different from those of P4 toxin respectively. The SH14 toxin was shown to have immunological cross-reactivity about 50% with P4 toxin when examined by Western hybridization.

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동부(Vigna sinensis)의 국부병반을 이용한 Cucumber mosaic virus와 Broad bean wilt virus 2의 구별 (Discrimination of Cucumber mosaic virus and Broad bean wilt virus 2 Using Local Lesions on Vigna sinensis)

  • 배선화;김미순;정민영;권순배;류기현;김국형;최장경
    • 식물병연구
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    • 제12권3호
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    • pp.221-225
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    • 2006
  • 용담에서 전형적인 모자이크 증상을 나타낸 바이러스 이병주 및 황화증상을 보이는 식물체를 채집하였다. 이들 중, 모자이크 증상의 잎을 접종하여 전신감염된 Nicotiana benthamiana로부터의 접종원을 동부의 초생엽에 접종하였을 때, 2가지 형태의 괴사 국부병반이 형성되었다. 즉 하나는 작은 크기 pinpoint 형태의 국부 괴사병반(SNS)이며, 다른 하나는 SNS보다 약간 큰 halo 형의 국부 괴사 병반(LNS)으로 뚜렷하게 구분되었다. 한편 황화 병징의 용담으로부터 증식된 접종원은 동부의 초생엽에 LNS만 형성하였다. 동부에 형성된 SNS 및 LNS형 병반을 각각 단일병반 분리하여 N. bentamiana에 접종하였다. 그 결과 SNS를 접종한 N. bentamiana에서는 접종엽에 무병징, 상위엽에는 전형적인 모자이크 증상이 발현되었다. 반면 LNS를 접종한 식물체에서는 접종엽에 퇴록 윤문이 형성되었으며, 상위엽에는 괴저를 동반한 모자이크 증상이 발현되었다. 이들 SNS 및 LNS 병반으로부터 분리한 바이러스를 dsRNA, RT-PCR 및 혈청학적 유연관계 분석을 통하여 SNS 병반은 CMV에 의해서, LNS는 BBWV2에 의해서 유도된다는 것을 확인하였다. 이 연구의 결과로부터 자연계에서 CMV와 BBWV2가 복합감염되어 병징의 구분이 어려운 경우, 동부에 접종하여 유도된 병반을 이용하여 두 바이러스의 감염을 쉽게 구분할 수 있는 방법으로 활용할 수 있을 것으로 생각되었다.

Repurposing Screens of FDA-Approved Drugs Identify 29 Inhibitors of SARS-CoV-2

  • Ku, Keun Bon;Shin, Hye Jin;Kim, Hae Soo;Kim, Bum-Tae;Kim, Seong-Jun;Kim, Chonsaeng
    • Journal of Microbiology and Biotechnology
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    • 제30권12호
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    • pp.1843-1853
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    • 2020
  • COVID-19, caused by the novel coronavirus SARS-CoV-2, has spread globally and caused serious social and economic problems. The WHO has declared this outbreak a pandemic. Currently, there are no approved vaccines or antiviral drugs that prevent SARS-CoV-2 infection. Drugs already approved for clinical use would be ideal candidates for rapid development as COVID-19 treatments. In this work, we screened 1,473 FDA-approved drugs to identify inhibitors of SARS-CoV-2 infection using cell-based assays. The antiviral activity of each compound was measured based on the immunofluorescent staining of infected cells using anti-dsRNA antibody. Twenty-nine drugs among those tested showed antiviral activity against SARS-CoV-2. We report this new list of inhibitors to quickly provide basic information for consideration in developing potential therapies.