• Title/Summary/Keyword: Rhabdovirus

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Optimization of a Diagnostic DNA Chip for Fish Rhabdovirus

  • Kim Young Ju;Kang Ji Hee;Kim Su Mi;Park Soo Il;Kim Sang Bong;Lee Myung Suk
    • Fisheries and Aquatic Sciences
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    • v.8 no.3
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    • pp.122-127
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    • 2005
  • A DNA chip that rapidly and accurately detects the viral genes in rhabdovirus-infected fish was developed. The N, Ml, and G proteins of three rhabdovirus strains, infectious hematopoietic necrosis virus (IHNV), viral hemorrhagic septicemia virus (VHSV), and flounder rhabdovirus (HIRRV), were selected for use as probes. The sequences of the corresponding genes were obtained, and probes were prepared by PCR using specific primer sets. The specificity of the probes was confirmed by cross PCR. The prepared probes were spotted on poly-L-lysine- or aminosilane-coated glass slides and hybridized with target DNA under several different conditions in order to determine the optimal hybridization temperature, glass-slide coating, and target cDNA concentration.

Isolation of rhabdovirus-like from fry of the fry of the snakehead fish, Channa arga (가물치, Channa arga 자어에서 분리한 Rhabdovirus 유사 병원체)

  • Kim, Su-Mi;Hong, Mi-Ju;Park, Su-Il
    • Journal of fish pathology
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    • v.19 no.1
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    • pp.55-64
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    • 2006
  • Rhabdovirus-like virus were isolated from the fry (15~30 days post hatching, dph) and rearing water of the snakehead fish Channa arga exhibiting mass mortality in spring of 2003 and 2004 in Korea. The isolates were propagated in EPC and SSN-1 cells but not replicated in FHM cells. The bullet-shaped viral particles (45×100 nm) appeared to be compact and a similar morphology to those of the rhabdoviruses in the infected EPC cells. The optimum temperature for virus replication was 20 to 25℃ but they could not replicate at 15℃. The isolates ShFRV-3 and ShFRV-5 from snakehead fish showed high pathogenicity against the fry (15 dph) and fingering (40 dph) of snakehead fish but did not in the larger size (90 dph).

Apoptosis in CHSE-214 Chinook Salmon Embryo Cells Infected with Hirame Rhabdovirus (HIRRV)

  • Sung Han-Gi;Kim Yeong-Jin;Jung Sung-Ju;Choi Won-Chul;Jung Tae-Sung;Choi Tae-Jin;Oh Myung-Joo
    • Fisheries and Aquatic Sciences
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    • v.5 no.1
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    • pp.32-35
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    • 2002
  • In this study, we investigated the mechanism of cell death in rhabdovirus-infected cells, chinook salmon embryonic cell line (CHSE-2l4) infected with hirame rhabdovirus (HIRRV). Studies using light microscopy, fluorescence microscopy, TUNEL method, electron microscopy, and agarose gel electrophoresis revealed changes in the cell morphology and DNA fragmentation indicative of apoptosis in early infection. It was observed that HIRRV induced apoptosis as well as necrosis in infected cells.

Generation of heterologous proteins-expressing recombinant snakehead rhabdoviruses (rSHRVs) using reverse genetics

  • Kwak, Jun Soung;Ryu, Sujeong;Kim, Ki Hong
    • Journal of fish pathology
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    • v.33 no.2
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    • pp.163-169
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    • 2020
  • Snakehead rhabdovirus (SHRV) is different from other fish novirhabdoviruses such as viral hemorrhagic septicemia virus (VHSV), infectious hematopoietic necrosis virus (IHNV), and hirame rhabdovirus (HIRRV) in that it replicates at high temperatures. Therefore, the delivery of foreign proteins to fish living at high water temperature would be possible by using recombinant SHRVs. In the present study, to evaluate the possible use of SHRV as a vehicle for foreign proteins delivery, we generated a recombinant SHRV that contains an enhanced-GFP (eGFP) gene between nucleoprotein (N) and phosphoprotein (P) genes (rSHRV-A-eGFP), and another recombinant SHRV expressing two heterologous genes by inserting an eGFP gene between N and P genes, and mCherry gene between P and M genes (rSHRV-AeGFP-BmCherry). Epithelioma papulosum cyprini (EPC) cells infected with the recombinant SHRVs showed strong fluorescence(s), suggesting the possible availability of recombinant SHRVs for the development of combined vaccines by expressing multiple foreign antigens.

A New Rhabdovirus (HRV-like) Isolated in Korea from Cultured Japanese Flounder Paralichthys olivaceus (양식산 넙치로부터 HRV-like Rhabdovirus의 분리)

  • Oh, Myung-Joo;Choi, Tae-Jin
    • Journal of fish pathology
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    • v.11 no.2
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    • pp.129-136
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    • 1998
  • In March 1997, a new rhabdovirus was isolated from moribund cultured Japanese flounder Paralichthys olivaceus in sea water tank and cage culture systems in Kyung-Nam and Chun-Nam province, Korea. At temperature $15^{\circ}C$ the virus replicated and induced cytopathic effects (CPE), which progressed to eventual cytolysis, in susceptible cell lines, including RTG-2 and EPC. The CHES-214 cell line was refractory. Virus particles were bullet-shaped and measured $70nm{\times}100$ to 150 nm in size. The isolate was sensitive to pH 3, to diethyl ether, and to heat ($50^{\circ}C$ 5 min, $60^{\circ}C$ 1 min). Viral replication was not inhibited by $10^{-4}$ M 5-iododeoxyuridine. Virus infectivity was reduced by anti-HRV (8401-H) rabbit serum, but can not reduced by antisera against infectious hematopoietic necrosis virus (IHNV), chum salmon reovirus (CSV), retrovirus of salmonid (RVS) and infectious pancreatic necrosis virus (IPNV). HRV virus antigen was detected by fluorescent antibody test (FAT) in the cytoplasm of infected EPC cell. Purified isolates virions were composed of: polymerase (L), glycoprotein (G), nucleoprotein (N) and 2 matrix proteins (M1 and M2). Based upon their relative mobilities, the estimated molecular weights of the proteins were: L, 160 kDa; G, 55 kDa; N, 45 kDa; M1, 26 kDa; and M2, 22 kDa.

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Protective immunogenicity of the G protein of hirame rhabdovirus (HIRRV) in flounder using DNA vaccine

  • Seo, Ji-Yeon;Kim, Ki-Hong;Kim, Sung-Koo;Kim, Young-Tae;Park, Tae-Jin
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2003.05a
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    • pp.313-314
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    • 2003
  • Antiviral DNA vaccine carrying a gene for a major antigenic viral protein have received considerable attention as a new approach in vaccine development. For fish viruses effects of DNA vaccine encoding viral G gene of infectious hematopoietic necrosis virus(IHNV) and viral hemorrhagic septicemia virus (VHSV)have been demonst.ated previously(Lapatra et al., 2001) Hirame rhabdovirus (HIRRV) causes hemorragic disease on flounder. (omitted)

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Detection of Fish Rhabdoviruses using a Diagnostic Fish Rhabdovirus DNA Chip

  • Kim, Young-Ju;Lee, Myung-Suk
    • Fisheries and Aquatic Sciences
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    • v.8 no.3
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    • pp.185-187
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    • 2005
  • We tested the in vivo ability of a DNA chip to detect virus-specific genes from virus-infected olive flounder Paralichthys olivaceus and rainbow trout Oncorhynchus mykiss. Target cDNA was obtained from total RNA of virus infected cell lines by reverse transcription (RT) and was labeled with fluorescent dye (Cy5-dUTP). The results show the successful detection of infectious hematopoietic necrosis virus (IHNV) and viral hemorrhagic septicaemia virus (VHSV) genes in the virus-infected fishes.

Molecular Cloning and Nucleotide Sequence of the G protein of a Korean Isolate of Infectious Hematopoietic Necrosis Virus (한국에서 분리된 IHNV-PRT의 G protein의 유전자 클로닝과 염기서열 분석)

  • 김영조;허강준;박정우;박정문
    • Korean Journal of Microbiology
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    • v.35 no.3
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    • pp.226-230
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    • 1999
  • To characterize the Korean isolate of infeciious hematopoietic necrosis virus (IHNV-PRT), a partial DNA fragment G gene of the MNV-PRT was amplified by RT-PCR. cloned inlo pGEM-T easy vector and analyzed for nucleotlde sequences. The size of the PCR pmduct was about 442 bp. The nucleotlde sequence homologies ofthe G gene of IHNV-PRT were 95%, 94%, 94% 94%, 93%, 53%. respectively. with those of foreign isolates of IHNV, IHNV-RB-76. IHNV-LR-73, MNV-K, IHNV-WRAC, Im-SRCV, IHNV-Col-85. However, it showed 81% homology with that of other fish rhabdovirus, hisame rhabdovirus (HRV). Frou~ the rcsults of deduced amino acid sequence homology analysis. G protein of IHNV-PRT showed 96% hornologies with those of foreign isolates of IHNV but 89% homology with that of HRV These results indicaled that, even though G gene of IHIW-PRT showed low homology with that of HRY it was highly conserved among different strains of THNV.

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Molecular Cloning and Nucleotide Sequence of the N Protein of a Korean Isolate of Infectious Hematopoietic Necrosis Virus (한국에서 분리된 전염성 조혈괴저바이러스의 N 단백질의 유전자 클로닝과 염기서열 분석)

  • Mun, Chang-Hoon;Kim, Hyun-Ju;Park, Jeong-Min;Cho, Wha-Ja;Cha, Seung-Ju;Yoon, Won-Joon;Park, Jeong-Jae;Lee, Eun-Hee;Kang, Hoe-Sung;Kim, Han-Do;Park, Jeong-Woo
    • Korean Journal of Microbiology
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    • v.34 no.1_2
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    • pp.69-73
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    • 1998
  • We have cloned and analyzed cDNA coding for nucleocapsid protein N from infectious hematopoietic necrosis virus(IHNV), IHNV-PRT, which was isolated in Korea. The N gene had open reading frame of 1,176 bp that encoded a 391 amino acids with a molecular weight of 42.3 kDa. The deduced amino acid sequence of N protein was 75-90% identical to those of foreign isolates, IHNV, but was 43% and 38% identical to those of other species of fish rhabdovirus, hirame rhabdovirus(HRV) and viral hemorrahagic septicemia virus(VHSV), respectively. However, it revealed high levels of sequence identity between 214-265 amino acid sequences among all species of fish rhabdovirus.

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