• Title/Summary/Keyword: CHSE-214 cell line

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A Study on infection symptom of infectious pancreatic necrosis virus(IPNV) in chinook salmon embryo cell line (연어 세포주의 전염성 췌장괴사 바이러스의 감염 증상에 관한 연구)

  • Kim, Young-Gill;Lee, Keun-Kwang;Chung, Ee-Yung
    • Journal of fish pathology
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    • v.5 no.1
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    • pp.1-7
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    • 1992
  • CHSE(Chinook Salmon Embryo)-214 fish cell lines was cultured in Eagle's minimal medium (MEM) supplemented with 10% fetal bovine serum and 2mM-glutamin. Optimum growth temperature of CHSE-214 cell line was $20^{\circ}C$. Infectious pancreatic necrosis virus(IPNV) was successfuly multiplied and showed the cytopathic effect in CHSE-214 cell line. Infection symptom of IPNV was observed with inverted phase contrast microscopy. At 6h-12hrs post-infection, the cells infected with IPNV were similer to normal cells. At 18-24hrs post-infection, the cells were somewhat round form and a little swollen form than normal cells. At 30hs post-infection, the cells were becoming more abnormal cells. At 48-68 post-infection, the infected cells were lysed and showed the severe cytopathic effect.

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Marine birnavirus (MABV)'s 5' terminal region of segment A acts as internal ribosome entry site (IRES)

  • Kim, So Yeon;Kim, Ki Hong
    • Journal of fish pathology
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    • v.34 no.1
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    • pp.17-22
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    • 2021
  • Eukaryotic translation is initiated by either cap-dependent or cap-independent way, and the cap-independent translation can be initiated by the internal ribosomal entry site (IRES). In this study, to know whether the 5'UTR leader sequence of marine birnavirus (MABV) segment A and segment B can act as IRES, bicistronic vectors harboring a CMV promoter-driven red fluorescent gene (mCherry) and poliovirus IRES- or MABV's leader sequence-driven green fluorescent gene (eGFP) were constructed, then, transfected into a mammalian cell line (BHK-21 cells) and a fish cell line (CHSE-214 cells). The results showed that the poliovirus IRES worked well in BHK-21 cells, but did not work in CHSE-214 cells. In the evaluation of MABV's leader sequences, the reporter eGFP gene under the 5'UTR leader sequence of MABV's segment A was well-translated in CHSE-214 cells, indicating 5'UTR of MABV's segment A initiates translation in the cap-independent way and can be used as a fish-specific IRES system. However, the 5'UTR leader sequence of MABV's segment B did not initiate translation in CHSE-214 cells. As the precise mechanism of birnavirid IRES-mediated translation is not known, more elaborate investigations are needed to uncover why the leader sequence of segment B could not initiate translation in the present study. In addition, further studies on the host species range of MABV's segment A IRES and on the screening of other fish-specific IRESs are needed.

Expression of the Heat Shock Proteins in HeLa and Fish CHSE-214 Cells Exposed to Heat Shock (어류 CHSE-214와 인간 HeLa 세포에서의 열충격에 의한 Heat Shock Protein의 발현)

  • 공회정;강호성김한도
    • The Korean Journal of Zoology
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    • v.39 no.2
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    • pp.123-131
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    • 1996
  • In this study, we examined the expression of heat shock proteins (HSPs) in fish cell line CHSE-2lnl and human HeLa cells exposed to heat shock. In fish CHSE-214 cells HSP70 was the major polvpeptide induced by an elevated temperature or an amino acid analog, while in HeLa cells HSP90 as well as HSP70 were prominently enhanced in response to these stresses. Pretreatment of actinomvcin D prior to heat shock completely inhibited the induction of fish HSP70, indicating the transcriptional regulation of fish HSP70 gene expression. In HeLa and CHSE-214 cells either recovering from heat shock or experiencing prolonged heat shock, attenuation in the HSP90 a'nd HSP70 induction occurred but both induction and repression of HSP70 synthesis appear 19 precede those of HSP90. Moreover, attenuation did not occur in the syntheses of 40 kDa and 42 kOto proteins which were only induced in CHSE-214 cells. The enhanced syntheses of these he proteins continued as long as CHSE-214 cells were Siven heat shock. These results suggest that down-regulation of HSP syntheses during prolonged heat shock may be controlled by several different. as vet undefined, mechanisms.

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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.

Activity Analysis of Misgurnus mizolepis Experssion Vector (미꾸라지 발현백터의 활성도 조사)

  • 함경훈;임학섭;황지연;박진영;김무상;이형호
    • Journal of Aquaculture
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    • v.11 no.4
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    • pp.457-463
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    • 1998
  • An expression vector, pUC19N6-luc, containing nuclear matrix attachment region(MAR) isolated from Misgurnus mizolepis liver and control expressino vector, pUC19-luc, were constructed. After these vectors were transferred into CHSE-214 cell line by electroporation, the expression rate of luckferase gens, copy number of vectors and chromosome integration of vectors were analyzed by using assay of luciferase activity, PCR and Southern blotting. While the expression pattern of luciferase gene of pUC19-luc was shown in typicla transient ecpression pattern, that of pUC19N6-luc was highly increased at the 5 days after transfectrion. Although the cope number of pUC19N6-luc vector was higher than that of pUC19-luc vector, these vectors were integrated into chromosome at the same time point in the transfected CHSE-214 cells. In conclusion, the increase of luciferase gene expression of pUC19N6-luc was resulted from not the maintaining of the high copy number but the formation of transcription-favorable structure by MAR effect after chromosomal integration.

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A Basical Study on Viral Haemorrhagic Septicemia of Rainbow trout, Salmo gairdneri (무지개송어의 바이러스성 출혈성 패혈증에 관한 기초적 연구)

  • Lee, Keun-Kwang
    • Journal of fish pathology
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    • v.7 no.1
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    • pp.7-11
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    • 1994
  • The viral hemorrhagic septicemia of rainbow trout, Salmo gairdneri was studied. The hematocrit values of diseased fish were very low than those of normal fish. And, the GOT and GPT values of serum of diseased fish were a little high than those of normal fish. Cytopathic effect of viral agents(serum and organs of diseased fish) was observed with inverted phase contrast microscopy. After 24hrs infection, the cells were showed the cytopathic effect.

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Toxicity of Disinfectants in Flounder Paralichthys olivaceus, Black Rockfish Sebastes pachycephalus and Black Sea Bream Acanthopagrus schlegelii (시판 소독제의 넙치 Paralichthys olivaceus, 조피볼락 Sebastes pachycephalus, 감성돔 Acanthopagrus schlegelii에 대한 독성)

  • Park, Kyung-Hee;Kim, Seok-Ryel;Kang, So-Young;Jung, Sung-Ju;Oh, Myung-Joo
    • Journal of Aquaculture
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    • v.21 no.1
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    • pp.7-12
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    • 2008
  • Toxicity of ten commercial disinfectants(hydrogen peroxide, sodium hypochlorite, chlorine dioxide, povidon iodine, formaldehyde, glutaraldehyde, quaternary ammonium compounds(QAC), didecyl dimethyl ammonium chloride(DDAC), ortho-dichlorobenzene, and copper sulfate) was measured by chinook salmon embryo-214 cell line and three fish species: flounder Paralichthys olivaceus, black rockfish Sebastes pachycephalus, and black sea bream Acanthopagrus schlegelii. The $LC_{50}$ levels of 24 hours acute toxicity with the ten disinfectants were tested in three species of fish. Effectiveness of ten chemical disinfectants were varied by each chemical as well as by species. Hydrogen peroxide showed the higest activity at 201, 269, and 139 ppm in the flounder, the black rockfish, and black sea bream, respectively. DDAC showed the lowest activity at 2.1, 1.0, and 1.5 ppm in the flounder, the black rockfish, and black sea bream, respectively. The highest variation was observed in copper sulfate by both the chemicals and the species.

Relationship between Viral Propagation and Apoptosis after Marine Birnavirus (MABV) Infection

  • Kim Yeong Jin;Choi Won Chul;Kim Hyeung Rak;Jung Sung Ju;Jung Tae Sung;Kim Jae Ho;Yeo In Kyu;Oh Myung Joo
    • Fisheries and Aquatic Sciences
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    • v.3 no.1
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    • pp.49-51
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    • 2000
  • This study was performed to confirm the relationship between viral propagation and apoptosis by the infection of marine birnavirus strain (MABV NF-4) on chinook salmon embryo (CHSE-214) cells. After 6 hr viral infection, MABV was detected by PCR method. Also, as a result of DNA assay on the cells, MABV infection resulted in a typical feature of apoptosis, DNA fragmentation. The results suggest that MABV replicated to high concentrations during the early stage of infection induces apoptosis.

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Effects of substitution of viral hemorrhagic septicemia virus genotype IVa glycoprotein with vesicular stomatitis virus (VSV) glycoprotein on cell line preference

  • Kim, Min Sun;Choi, Tae-Jin;Kim, Ki Hong
    • Journal of fish pathology
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    • v.30 no.2
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    • pp.71-78
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    • 2017
  • The glycoprotein of novirhabdoviruses is known to play a critical role in the determination of host specificity. Viral hemorrhagic septicemia viruses (VHSVs) in different genotypes have different glycoprotein sequences and show different preferences for specific cell lines. In this study, to know whether the glycoprotein is solely responsible for the host cell preference of VHSV, a recombinant VHSV expressing vesicular stomatitis virus (VSV) glycoprotein instead of VHSV IVa glycoprotein (rVHSV-VSV-G) was generated by reverse genetics and inoculated into several fish cell lines, then, cytopathic effect (CPE) and viral growth caused by rVHSV-VSV-G infection were compared with those caused by rVHSV-wild that was previously generated and has the same genomic sequence with wild-type VHSV except a few nucleotides. The plaque numbers of rVHSV-VSV-G were significantly higher in EPC, BF-2 and GF cells than those of rVHSV-wild. However, in HINAE cells (originated from olive flounder), rVHSV-VSV-G titer was significantly lower than rVHSV-wild titer, and both recombinant VHSVs were not grown well in CHSE-214 cells. Although statistical significances were detected in the titers between rVHSV-wild and rVHSV-VSV-G in several cell lines, the cell line-preference order of rVHSV-VSV-G was not different from that of rVHSV-wild. These results suggest that the replacement of VHSV glycoprotein may not completely change host cell preference, and other regions of VHSV might also involve in the determination of host cell preference.

Studies on Viral Disease of masu salmon, Oncorhynchus masou-II Isolation of infectious hematopoietic necrosis virus form masu salmon fry (산천어의 바이러스성 질병에 관한 연구-II -산천어 치어에서 1HNV 분리-)

  • Sohn, Sang-Gyu;Park, Myoung-Ae;Park, Jeong-Woo
    • Journal of fish pathology
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    • v.6 no.2
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    • pp.87-92
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    • 1993
  • In February of 1990, an epizootic disease to masu salmon. Onchorynchus masou cultured at the hatchery of trout in Samchuk. Kwangwondo have broken out and induced heavy mortality. An infectious hematopoietic necrosis virus(IHNV) was isolated from diseased masu salmon fry by the use of fish cell line, CHSE-214. This IHNV isolated from masu salmon was compared with USA isolates of IHNV, SRCV and RB-76 by analysis of virion proteins in sodium dodecyl sulfate poly-acrylamide gel electrophoresis (SDS-PAGE) and neutralization tests with two monoclonal antibodies raised against SRCV(MAb SRCV/A4) and RB-76(MAb RB/B5). In the antigenicity and the size of structural proteins. this IHNV, SCS atrain was smilar to RB-76 belonged to the electropherotype I proposed by Hsu et al.(1986).

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