• Title/Summary/Keyword: Virus concentration

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Effect of Fumonisin B1 on the Bacterial Virus Multiplication (세균 바이러스 증식에 대한 Fumonisin B1의 영향)

  • 이길수
    • Toxicological Research
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    • v.12 no.1
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    • pp.17-20
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    • 1996
  • The effect of Fumonisin B1, a mycotoxin produced by Fusarium moniliforme on bacterial viruses P1 and Lambda, was investigated by the virus plaque assay. Fumonisin B1 inhibited the P1 viral multiplication in the concentration range from $100{\mu}g$/ml to $400{\mu}g$/ml. The inhibition was Fumonisin B1 concentration-dependent. Another bacterial virus Lambda multiplication was also inhibited by lower concentration of Fumonisin B1 ($10{\mu}g$/ml~$50{\mu}g$/ml). This inhibition was dependent on Fumonisin B1 and on virus-Fumonisin B1 reaction time. Sensitivity of bacteriophage Lambda to Fumonisin B1 was higher than that of P1 virus. Lambda vital DNA was treated in vitro with Fumonisin B1 at various concentration. Significant DNA fragmentation by Fumonisin 191 was observed in the agarose gel electrophoresis. Lambda viral DNA was partially digested even in the Fumonisin B1 $10{\mu}g$ and the level of its fragmentation was dependent on Fumonisin B1 amount up to $30{\mu}g$ per assay.

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Gene Transfer into Chicken Embryos using Defective Retroviral Vectors Packaged with Vesicular Stomatitis Virus G Glycoprotein Envelopes (Vesicular Stomatitis Virus G Glycoprotein Envelope으로 포장된 Defective Retroviral Vector를 이용한 닭의 배로의 유전자 전이)

  • 권모선;임은정;허영태;이훈택;이영만;김태완
    • Korean Journal of Animal Reproduction
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    • v.25 no.2
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    • pp.171-180
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    • 2001
  • Compared to other gene transfer system, the advantages of retrovirus-mediated gene transfer are technical ease, efficient expression and genetic stability. Despite the high potency of the retrovirus vector system in gene transfer, one of the drawbacks is a difficulty in concentration of virus stock. To overcome this problem, we tested a new retrovirus vector system producing the progeny retrovirus particles encapsidated with VSV-G (vesicular stomatitis virus G glycoprotein). The infectivity of this virus was not sacrificed by ultracentrifugal concentration and the host cell range extended from all mammalian to fish embryos. Virus titer after 1,000 x concentration was more than 10$^{8}$ CFU/ $m\ell$ on most of the target cell lines. We applied this pantropic viruses in transgenic chicken production by injecting the concentrated (100$\times$) stock into subgerminal cavity of stage X chicken embryos. The survival rate of chicken embryos after injection was about 20% and gene integration rate in surviving embryos was scored almost 100%. Analyses of RT-PCR and fluorescence microscopy, however, showed no evidence of the transgene expression.

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Studies on the Pathogenicity of Cytoplasmic Polyhedrosis Virus and the Phenomena of Induction and Interference by Oral Inoculation of Foreign Virus to the Silkworm, Bombyx mori L. (가잠에 있어서 세포질다각체 바이러스의 병원성 및 이형 바이러스 접종에 의한 유발 간섭현상에 관한 연구)

  • 김근영;문재유;임종성;정태암
    • Journal of Sericultural and Entomological Science
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    • v.17 no.2
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    • pp.101-118
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    • 1975
  • The pathogenicity of hexagonal and tetragonal cytoplasmic polyhedron virus to one of present leading silkworm varieties, Jam 103${\times}$Jam 104, and its parents, Jam 103 and Jam 104, was investigated. The activation of occult virus by oral inoculation of foreign virus as well as the interference phenomena between the activated and inoculated cytoplasmic polyhedron viruses was observed and the results obtained are as follows. 1. The pathogenicity of cytoplasmic polyhedrosis viruses. 1) There was a high significant difference in the pathogenicity of hexagonal polyhedron virus between the hybrid and its parents showing the lowest infection rate in the hybrid (Jam 103${\times}$Jam 104), medium infection rate in the Japanese line (Jam 103) and the highest infection rate in the Chinese line (Jam 104). In the pathogenicityof tetragonal polyhedron virus, a significant difference was observed only between the hybrid (Jam 103${\times}$Jam 104) and the Chinese line (Jam 104) by showing a higher infection rate in the Chinese line than in the hybrid. 2) The pathogenicity of both hexagonal and tetragonal polyhedron viruses showed a high significant difference in different silkworm instars inoculated by showing a higher infection rate at the second instar than at the fourth instar. 3) The pathogenicity of both hexagonal and tetragonal polyhedron viruses was increased as the concentration of viruses inoculated increased. 2. The phenomena of induction and interference by oral inoculation of foreign virus. 1) The induction rate of cytoplasmic polyhedrosis virus was higher in the parents than in the hybrid. In the parents. a higher rate in the Chinese line than in the Japanese line was observed. 2) The effect of inoculation at different instars on the induction was studied and the induction rate was higher at the second instar inoculated than at the fourth instar inoculated. 3) The degree of activation of hexagonal polyhedron virus with inoculation of tetragonal polyhedron virus was very high when a lower concentration of virus was inoculated and it was very low when a higher concentration of virus was inoculated 4) The degree of activation of tetragonal polyhedron virus with inoculation of hexagonal polyhedron virus was very low when a lower concentration of virus was inoculated and it was not observed when a higher concentration of virus was inoculated. 5) The mixed infection rate with hexagonal and tetragonal polyhedron viruses was higher at the second instar inoculated than at the fourth instar inoculated. 6) It was observed that the secondary hexagonal pc]yhedron virus activated interfered with the primary tetragonal polyhedron virus inoculated when the inoculated concentration of the primary virus is low and the primary virus inoculated interfered with the secondary virus activated when the inoculated concentration of the primary virus is high.

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Development of a Virus Concentration Method and its Application for the Detection of Noroviruses in Drinking Water in China

  • Liu, Junyi;Wu, Qingping;Kou, Xiaoxia
    • Journal of Microbiology
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    • v.45 no.1
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    • pp.48-52
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    • 2007
  • A new procedure for the concentration of nonoviruses from water samples has been developed. This procedure (calcium flocculation-citrate dissolution method) uses the following steps: virus flocculation formed by treatment with 1 M $CaCl_2$ and 1 M $Na_2HPO_4$, virus release by sodium citrate dissolution (0.3 M Na citrate, pH 3.5), and virus re-concentration by ultrafiltration. When reverse transcription (RT)-PCR was performed after the procedure, the overall detection sensitivity for seeded noroviruses in a one liter drinking water sample was as low as 1 RT-PCR unit, which is equal to a $10^{-6}$ dilution of the virus sample. This approach showed at least a 5-fold-higher sensitivity than the current method with its three steps of adsorption-elution-concentration. The newly developed procedure was used to test different brands of bottled drinking water from China for putative contamination with noroviruses. A total of 144 samples were analyzed; all of the samples were negative for norovirus specific nucleic acids.

Titer Amplification of GALV (Gibbon Ape Leukemia Virus) Pseudotyped Retrovirus Vectors Produced from PG13 Cells (PG13 Cell로부터 생산된 GALV (Gibbon Ape Leukemia Virus)-pseudotyped Retrovirus Vector의 증폭)

  • 김태완;박윤엽;권모선;염행철;김경화;박영식;박세필
    • Korean Journal of Animal Reproduction
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    • v.21 no.4
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    • pp.397-403
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    • 1997
  • For the ultimate goal of efficient retrovirus vector-mediated transgenic animal production, we tried to increase virus titer by employing three methods: boosting virus production by treating virus-producing cells with sodium butyrate, concentration of virus stock by either filtration or ultracentrifugation. Compared to the control, applications of sodium butyrate (5 mM) treatment and filtration resulted in only 3 and 3. 6 folds of titer increases on bovine EBTr target cells, respectively. However, concentration of virus-containing medium by ultracentrifugation showed 12.5 folds of titer increase compared to the control (10${\times}$10$^5$ LacZ$^+$ TU Im), indicating the best method which can enhance retrovirus vector-mediated transgenic animal production.

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Development of a Virus Elution and Concentration Procedure for Detecting Norovirus in Oysters

  • Ha, Sook-Hee;Woo, Gun-Jo;Hwang, In-Gyun;Choi, Weon-Sang
    • Food Science and Biotechnology
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    • v.18 no.5
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    • pp.1150-1154
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    • 2009
  • Low levels of virus contamination and naturally occurring reverse transcription-polymerase chain reaction (RT-PCR) inhibitors restrain virus detection in oysters. A rapid and efficient oyster-processing procedure that can be used for sensitive virus detection in oysters was developed. Poliovirus type 1 Sabin strain was used to evaluate the efficacy of virus recovery. The procedure included (a) acid-adsorption and elution with buffers (0.25M glycine-0.14 M NaCl, pH 7.5; 0.25M threonine-0.14M NaCl, pH 7.5); (b) polyethylene glycol (PEG) precipitation; (c) resuspension in Tween 80/Tris solution and chloroform extraction; (d) the second PEG precipitation; (e) viral RNA extraction with TRIzol and isopropanol precipitation; and (f) RT-PCR combined with semi-nested PCR. The overall recovery of elution/concentration was 19.5% with poliovirus. The whole procedure usually takes 19 hr. The overall detection sensitivity was 4 RT-PCR units of genogroup I norovirus (NoV) and 6.4 RT-PCR units of genogroup II Nov/25 g of oysters initially seeded. The virus-detecting method developed in this study should facilitate the detection of low levels of NoV in oysters.

Improvement of Virus Productivity by Sodium Butyrate in the Production of Porcine Transmissible Gastroenteritis Virus Vaccine (Sodium butyrate에 의한 돼지 전염성 위장염 바이러스 백신의 생산성 향상)

  • Lee, Chang-Jin;Kim, Cheol-Min;Jeong, Yeon-Ho
    • KSBB Journal
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    • v.26 no.2
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    • pp.107-111
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    • 2011
  • The essential operating parameters in virus vaccine production are multiplicity of infection (MOI), harvest time, and infection time. Stimulating agents also can be applied in order to improve vaccine productivity further. We investigated the optimum operating conditions in porcine transmissible gastroenteritis virus (TGEV) vaccine production and the applicability of sodium butyrate (NaBu) as a stimulating agents for the improvement of vaccine productivity. The optimum MOI, infection time, and harvest time for high production of TGEV by swine testicle (ST) cells were found to be 0.0001 pfu/cell, 3 day after cell inoculation, and 24 hpi, respectively. NaBu is known as a histone deacetylase inhibitor that has been widely used for the high expression of recombinant protein using mammalian cells and for the enhancement of virus propagation. So we tried to examine the potential of NaBu as a stimulating agent and to determine the optimum concentration by comparing TGEV titers with different range of NaBu concentration. TGEV titer with 5 mM NaBu was 1.5 times higher than control. Therefore, we concluded that NaBu can be a promising agent for stimulating various vaccine production including TGEV and the optimum NaBu concentration for TGEV production was determined to be 5 mM.

A tendency of herbal medicines on anti-viral effect (항바이러스 효과에 응용되는 한약의 연구 동향)

  • Lim, Sung-Woo
    • THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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    • v.4 no.1
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    • pp.199-209
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    • 1998
  • In order to find antiviral effect against Human immunodeficiency virus(HIV), Herpes simplex virus type I(HSV-1) and II(HSV-2) from herb medicines, publicated 29 paters on anti-viral effect of herbal medicines and a convenient virus-induced cytopathic effect (CEP) inhibition assay was introduced. The major virus on experiment are HIV, Hepatitis B virus and HSV-1,2. Those of other studies showed inhibition of infected virus DNA replication and screening test of herbal medicines. More than 15 extractions were prepared by pure water boiling from herbal medicines, and their toxicity of infected cell and anti-viral activities were evaluated. Among them, the major part of herbal medicines showed cell stability compared with the contrast. Cytotoxic concentration (CC) of the $H_2O$ extracts of Padoo against HIV was <4.0, Hyungbangpaedoksan against HIV was 9.3, Whangyonhaedoktang against HIV-1 and HSV-2 was 15.3. These are high level cytotoxic concentration compared with the contrast. But antiviral effect was unable to figure out for selective $index(SI)=CC_{50}/EC_{50}$. The other herbal medicines were unable to showed potent anti-HIV and anti-HSV activity. The antiviral activation using herbs in this thesis have unlimited objects, to select research object will help to show the direction of antiviral drug development that have less side effect and more excellent efficiency.

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Effect of Serum on Varicella-Zoster Virus Propagation (수두 바이러스 증식에 미치는 혈청의 영향)

  • 전복환;우규진
    • KSBB Journal
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    • v.9 no.3
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    • pp.272-278
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    • 1994
  • Attenuated varicella-zoster virus(VZV) was cultured in human embryonic lung cells. The effects of serum type and its concentration on the production of VZV were studied. Regardless of cell culture conditions, VZV yield was increased with multiplicity of infection, and the total cell concentration was decreased after virus infection. The newborn calf serum, calf serum, or horse serum was not as good as the fetal bovine serum or calf serum supplemented with iron for the propagation of VZV in the human embryonic lung cells. The yields of total VZV(cell-associated virus and cell-free virus) in the medium with calf serum containing iron were comparable to those in the medium supplemented with fetal bovine serum. Furthermore, some components of serum appear to be important for the maintenance of VZV infectivity.

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Production of Newcastle Disease Virus by Vero Cell Culture

  • Jeon, Ju-Mi;Jeon, Gye-Taek;Kim, Ik-Hwan;Lee, Sang-Jong;Jang, Yong-Geun;Jeong, Yeon-Ho
    • 한국생물공학회:학술대회논문집
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    • 2002.04a
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    • pp.271-272
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    • 2002
  • Newcastle disease virus (NDV) vaccines were produced from Vero cells by using lively attenuated virus strain. The MOI of 0.1.' serum concentration of 2%. initial pH of 8.0. and infection time of 3 days were found to be optimum conditions for vaccine production. The treatment of polycation enhanced the virus production. When ascorbic acid was added as an antioxidant, NDV production was also enhanced. Utilization of $CaCl_2$ showed an inhibitory effect on the propagation of NDV. It was also found the ammonium ion concentration higher than 4mM inhibited virus production. Thus ammonium ion removal system was tried for the efficient production of NDV vaccine.

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