• Title/Summary/Keyword: Viral density

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Development of Serum Free Medium and Optimization of Porcine Rotavirus Vaccine Production

  • Ko, Yun-Mi;Kim, Myoung-Hwa;Kim, Min-Young;Jeong, Yeon-Ho
    • 한국생물공학회:학술대회논문집
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    • 2005.04a
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    • pp.207-209
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    • 2005
  • Serum is a potential source of bacterial, mycoplasmal and viral contamination, and it has a possibility of the introduction of serum proteins, prion and pyrogens into the final vaccine product. For porcine Rotavirus vaccine production, it is necessary to develop serum free medium which do not cause those problems. A new serum free medium was developed for porcine Rotavirus vaccine based on DMEM, and the performance of developed serum free medium was evaluated in terms of Vero cell growth and Rotavirus vaccine production. The cell density, gown in serum free medium developed, was similar with that in serum supplemented medium. Also, it was higher than that in other commercially available serum free medium. The productivity of Rotavirus vaccine using serum free medium developed and optimum production strategies will be also discussed.

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Action of Ozone on Bacterial Virus f2 (세균성 Virus f2에 대한 Ozone의 불활성작용)

  • 김치경
    • Korean Journal of Microbiology
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    • v.18 no.3
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    • pp.123-132
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    • 1980
  • Bacterial virus f2 and its RNA were examined to elucidate the mode of ozone utilizing sucrose density gradient analysis and electtron microscopic techniques. the inactivation kinetics of the virus f2 by ozonation showed that the viruses were inactivated during the first 5 sec of the reaction and were further inactivated at a slower rate during the next 10 min at 0.09 and 0.8mg/l ozone concentrations. The virus coat was broken by ozonation into many pieces of protein subunits and the adsorption of the viruses to the host pili was inversely related to the extent of the breakage of the virus. The viral RNA was released from the virus particles during ozone, but ozone inactivation of the RNA enclosed in the protein coat could not ruled out the possibility that the RNA was secondarily sheared by a reaction with the broken coat protein.

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Monitoring of infectious diseases in three grouper species (Hyporthodus septemfasciatus, Epinephelus moara and E. moara ♀× E. lanceolatus ♂) cultured in recirculating aquaculture systems (순환여과시스템에서 양식되는 바리과 어류 3종[능성어, (Hyporthodus septemfasciatus), 자바리(Epinephelus moara), 교잡종 대왕자바리(E. moara ♀× E. lanceolatus ♂)]의 감염성 질병 모니터링)

  • Hee-Jae Choi;Da-Yeon Choi;SungHyun Jo;JeYun Shin;Jong Yeon Park;In-chul Bang;Yue Jai Kang
    • Journal of fish pathology
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    • v.37 no.1
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    • pp.97-110
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    • 2024
  • The Serranidae is high-quality fish species with good meat quality and is traded at high price, and is attracting attention in South Korea as a cultured species that creates high added value. However, the high-density fish farming for mass production increases the risk of mass mortality due to infectious diseases, leading to enormous economic losses. Therefore, in order to safely prevent and protect farmed fish from serious infectious diseases, it is necessary to conduct disease monitoring on a regular basis. In this study, Hyporthodus septemfasciatus, Epinephelus moara, and the hybrid longtooth grouper (E. moara ♀×E. lanceolatus ♂) were collected once a month from fish farm of Garorim and Aquabiotech Co., Ltd for a total of six months, from July to December 2023. We investigated infections of five species of bacterial diseases, including Flavobacterium columnare, six species of viral diseases, including LCDV (lymphocystis disease virus), and parasitic pathogens in grouper farms. As the result, Vibrio vulnificus and V. harveyi were detected in H. septemfasciatus in August, in the case of viral diseases, NNV was detected in H. septemfasciatus from July to August using RT-PCR or PCR. Finally, In the case of parasitic diseases, Tricodina sp. was detected in E. moara and the hybrid longtooth grouper from August to December.

Molecular biological characterization of the new virus isolated from abnormally swimming salmonid - Reverse transcriptase activity - (이상유영(異常遊泳) 증상(症狀)을 나타내는 연어과 어류(魚類)에서 분리(分離)된 새로운 바이러스의 분자생물학적(分子生物學的) 특성(特性) - 역전사효소 활성 -)

  • Oh, Myung-Joo
    • Journal of fish pathology
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    • v.9 no.2
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    • pp.177-183
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    • 1996
  • A new RNA virus isolated from abnormally swimming behavior has caused mortalities in salmonid fish (Oh et al., 1995 a), A reverse transcriptase (RTase) activity of the virus was determined by using poly r(A) : oligo d(T) as templete : primer. This RTase activity was associated with virus particles of buoyant density of 1.16 g/ml. The virus particles in sucrose fractions were enveloped and were about 85 nm diameter with central electron-dense core. The brain and kidney samples of artificially infected fish showed RTase activity. Virus particle associated proteins about 120, 80, 65, 61, 48, 42, 35, 30, 25, 19, and 15 kDa were observed when examined by polyacrylamide gel electrophoresis analysis. This study showed the presence of a new retrovirus in salmonid fish, which tentatively called RVS (Retrovirus of salmonid).

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Isolation and Characterization of the Smallest Bacteriophage P4 Derivatives Packaged into P4-Size Head in Bacteriophage P2-P4 System

  • Kim, Kyoung-Jin;Song, Jae-Ho
    • Journal of Microbiology
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    • v.44 no.5
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    • pp.530-536
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    • 2006
  • Bacteriophage P4, a satellite phage of coliphage P2, is a very useful experimental tool for the study of viral capsid assembly and cos-cleavage. For an in vitro cos-cleavage reaction study of the P2-P4 system, new shortened and selectable markers containing P4 derivative plasm ids were designed as a substrate molecules. They were constructed by swapping the non-essential segment of P4 DNA for either the kanamycin resistance (kmr) gene or the ampicillin resistance (apr) gene. The size of the genomes of the resulting markers were 82% (P4 ash8 delRI:: kmr) and 79% (P4 ash8 delRI:: apr) of the wild type P4 genome. To determine the lower limit of genome size that could be packaged into the small P4-size bead, these shortened P4 plasmids were converted to phage particles with infection of the helper phage P2. The conversion of plasmid P4 derivatives to bacteriophage particles was verified by the heat stability test and the burst size determination experiment. CsCl buoyant equilibrium density gradient experiments confirmed not only the genome size of the viable phage form of shortened P4 derivatives, but also their packaging into the small P4-size head. P4 ash8 delRI:: apr turned out to be the smallest P4 genome that can be packaged into P4-sized head.

Expression of Hepatitis C Virus Structural Proteins in Saccharomyces cerevisiae

  • LEE JONG-SOO;YU JUNG;SHIN HYUN-JIN;KIM YOUNG-SANG;AHN JEONG-KEUN;LEE CHONG-KIL;POO HARYOUNG;KIM CHUL-JOONG
    • Journal of Microbiology and Biotechnology
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    • v.15 no.4
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    • pp.767-771
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    • 2005
  • Expression in yeast may prove more amenable to generating large amounts of viral antigens for a vaccine candidate. We, therefore, cloned the gene encoding the Hepatitis C virus (HCV) structural proteins (C-El-E2, c740) fused in-frame with, and immediately 3' to, the chicken-lysozyme signal peptide (C-SIG) gene and under the control of the yeast glyceraldehyde-3-phosphate dehydrogenase gene promoter. In yeast, the HCV structural proteins were expressed in two different forms: a processed and a nonprocessed aggregated form. Biophysical characterization by sucrose linear gradient centrifugation revealed that both forms were present in the same fractions with a buoyant density of 1.127-1.176 g/$cm^3$. These findings suggest that the efficient synthesis of HCV structural proteins in yeast may be an important tool to study virus assembly and may lead to the development of an HCV vaccine.

Isolation and characterization of fowl adenovirus serotype 4 from chickens with hydropericardium syndrome in Korea

  • Park, Hong-Su;Lim, Il-Soo;Kim, Sang-Kyu;Kim, Toh-Kyung;Yeo, Sang-Geon
    • Korean Journal of Veterinary Research
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    • v.51 no.3
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    • pp.209-216
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    • 2011
  • Four strains of fowl adenovirus (FAdV) were isolated from 4 flocks of broiler or layer chickens affected by hydropericardium syndrome in Korea. These FAdVs were classified as serotype 4 by restriction fragment length polymorphism patterns of hexon genes and whole genomes. The virus exhibited cytopathic effects consisting of rounding, ballooning and clustering in primary chicken embryo liver cell cultures. In transmission electron microscopy, virus particles in hexagonal shape aggregated exclusively in the nuclei of hepatocytes of the chickens as the typical appearances in adenovirus infections. Buoyant density of the virus in cesium chloride (CsCl) was 1.34 g/mL. The virus was stable to chloroform, ether, 50~70% ethanol, acidic condition at pH 3, 0.25% trypsin (1 : 250), heat at $50^{\circ}C$ for 30 min, but labile to 100% ethanol, heat at $52{\sim}60^{\circ}C$ for 30 min, 1 M $MgCl_2$ at $50^{\circ}C$ for 1 h, 1 : 2,000 formalin (37%). All of the physicochemical properties pertained to the characteristics of adenoviruses. Eight viral polypeptides were determined in CsCl-purified virus by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

Production of Monoclonal Antibodies Against Nervous Necrosis Virus (NNV, RGNNV genotype) (신경괴사증바이러스(nervous necrosis virus, RGNNV genotype)에 대한 단클론 항체 생산)

  • Kim, Wi-Sik;Kim, Si-Woo;Oh, Myung-Joo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.51 no.3
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    • pp.328-331
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    • 2018
  • We developed and subsequently characterized mouse monoclonal antibodies (MAbs) against nervous necrosis virus (NNV, RGNNV genotype). We established six hybridoma clones secreting MAbs against NNV antigen: 2B1, 2B11, 2C12, 13C1-1, 13C1-2 and 14D11. All six MAbs belonged to the IgG2a isotype with a kappa light chain and their reactivity recognized against the 41 kDa coat protein of NNV by Western blot analysis. The affinity constants of the six MAbs were measured by enzyme-linked immunosorbent assay (ELISA). All six MAbs reacted with two NNV isolates (SgNag05 and Gemunodo06), while no reactivity was observed with five know fish viruses, namely marine birnavirus, infectious pancreatic necrosis virus, viral hemorrhagic septicemia virus, hirame rhabdovirus, and infectious hematopoietic necrosis virus. Moreover, high ELISA optical density (OD) values (0.87-1.42) were observed in the brain tissues of NNV-infected sevenband grouper, while low OD values (less than 0.12) were recorded in the brain tissues of uninfected fish. These results suggest that these six MAbs are highly competent and useful for the detection of NNV with the RGNNV genotype.

A Inquiry of Tracer Gas for Analysis of Dispersion and Prediction of Infection Possibility according to Airborne Viral Contaminants (건축공간에서 공기 감염균 확산을 해석하기 위한 추적가스 고찰과 농도에 따른 감염 위험성 예측 연구)

  • Lim, Tae-Seob;Kang, Seung-Mo;Kim, Byung-Seon
    • Korean Institute of Interior Design Journal
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    • v.18 no.3
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    • pp.102-113
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    • 2009
  • The SARS virus began to appear and spread in North America and Southeast Asia in the early 2000' s, infecting and harming many people. In the process of examining the causes for the virus, studies on the airborne SARS virus and the way it spread were carried out mainly in the medical field. In the field of architecture, studies were done on the diffusion of air pollutants in buildings using gases such as $CO_2$, $N_2O$, or $SF_6$, but research on virus diffusion was limited. There were also explanations of only the diffusion process without accurate information and discussion on virus characteristics. The aim of this study is to analyze the physical characteristics of airborne virus, consider the possibility of using coupled analysis model and tracer gas for analyzing virus diffusion in building space and, based on reports of how the infection spread in a hospital where SARS patients were discovered, analyze infection risk using tracer gas density and also diffusion patterns according to the location, shape, and volume of supply diffusers and exhaust grilles. This paper can provide standards and logical principles for evaluating various alternatives for making decisions on vertical or horizontal ward placement, air supply and exhaust installation and air volumes in medium or high story medical facilities.

Biochemical and Immunological Characterization of the DNA Polymerase and RNase H in Feline Leukemia Virus (고양이 백혈병 바이러스의 DNA Porymerase와 RNase H의 생화학적 및 면역학적 연구)

  • Park, Hyune-Mo
    • The Korean Journal of Zoology
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    • v.22 no.4
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    • pp.141-152
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    • 1979
  • Feline leukemia virus DNA polymerase was purified by ion-exchange and nucleic acid affinity chromatographies. The enzyme consists of a single polypeptide chain of approximately 72, 000 molecular weight as determined by both of a glycerol density gradient centrifugation and SDS-polyacrylamide gel electrophoresis. The preferred divalent cation for DNA synthesis is $Mn^2+$ on a variety of template-primers, and its optimum concentration appears to be significantly lower than reported results of other mammalian type-C viral enzymes. The divalent cation requirement for maximum activity of RNase H is similar to those of DNA polymerase. Both DNA polymerase and RNase H activities appear to reside on the same molecule as demonstrated by the copurification of both activities through various purification steps. An additional RNase H without detectible polymerase activity was generated by a limited chymotrypsin digestion. This RNase H activity was inhibited equally effectively as RNase H in the intact reverse transcriptase by antisera prepared against reverse transcriptase of feline leukemia virus. Neutralization and binding test showed that antibody binding to reverse transcriptase molecule did not completely inhibit the polymerase activity.

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