• Title/Summary/Keyword: virions

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A Stable Preservation of Extracellular Nonoccluded Virions from Autographa californica Nuclear Polyhedrosis Virus Infection (Autographa californica 핵다면체 바이러스의 세포외 미봉입비리온의 안전한 보존)

  • 이형환;이근광;이건주
    • Microbiology and Biotechnology Letters
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    • v.18 no.6
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    • pp.660-661
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    • 1990
  • A stable preservation method of extracellular non-occluded virion of Autographa californica nuclear polyhedrosis virus (AcNPV) was studied. AcNPVL-1 strain infected to Spodoptera frugiperda cell line and then the culture media were centrifuged. After centrifugation the supernatant containing extracellular nonoccluded virions of the AcNPV was harvested and incubated at $4^{\circ}C$ . Even after the extracellular nonoccluded virions were incubated at $4^{\circ}C$ for about 11 years, the infectivity and multiplication property of the nonoccluded virions in the S. frugiperda cell line were normal. However the titers of the nonoccluded virions in TC-100 medium measured about 11 years ago decreased from $8.9 \times 10^7\; to \;3.8 \times 10^5$ pfu per ml. The AcNPV genome DNA fragment patterns from digestion with Hind11 and EcoRI restriction endonucleases did not change. The AcNPV nonoccluded virions were stable at $4^{\circ}C$ in the cultured medium of more than 10 years and the preservation of AcNPV nonoccluded virions at $4^{\circ}C$ is easy and useful for handling.

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Identification of a novel type of small molecule inhibitor against HIV-1

  • Kim, Byung Soo;Park, Jung Ae;Kim, Min-Jung;Kim, Seon Hee;Yu, Kyung Lee;You, Ji Chang
    • BMB Reports
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    • v.48 no.2
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    • pp.121-126
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    • 2015
  • Here we report a new chemical inhibitor against HIV-1 with a novel structure and mode of action. The inhibitor, designated as A1836, inhibited HIV-1 replication and virus production with a 50% inhibitory concentration ($IC_{50}$) of $2.0{\mu}M$ in an MT-4 cell-based and cytopathic protection antiviral assay, while its 50% cytotoxic concentration ($CC_{50}$) was much higher than $50{\mu}M$. Examination of the effect of A1836 on in vitro HIV-1 reverse transcriptase (RT) and integrase showed that neither were molecular targets of A1836. The characterization and re-infection assay of the HIV-1 virions generated in the presence of A1836 showed that the synthesis of early RT products in the cells infected with the virions was inhibited dose-dependently, due in part to abnormal protein formation within the virions, thus resulting in an impaired infectivity. These results suggest that A1836 might be a novel candidate for the development of a new type of HIV-1 inhibitor.

Inactivation by Chemical Disinfectants in vitro against Tobacco Mosaic Virus (화학적 제어제에 의한 담배모자이크 바이러스의 불활성화)

  • Choi, C.W
    • The Journal of Natural Sciences
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    • v.10 no.1
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    • pp.17-21
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    • 1998
  • Numerous chemicals were tasted to show antiviral activity in vitro against tobacco mosaic virus (TMV). With a brief exposure of TMV to 1 N HCl or 1-0.1 N NaOH, Virions and their encapsidated RNAs were degraded completely and rapidly. When TMV was exposed to 0.1 N HCl, the hydrolysis of viral capsid in 5 min after treatment was observed in the 1% agarose gel. Virions and their encapsidated RNAs were not degraded by 0.01N HCl of 0.01N NaOH. These characteristics indicate that a short exposure to optimal concentration of acid or base is of practical value in eliminating infectious virus. The treatment of 50% isopropanol or UV light did not damage in viral integrity or their encapsidated RNAs. Disinfection of the agricultural tools and laboratory equipments using appropriate disinfectants is necessary to prevent cross contamination if farm and laboratory.

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Characterization of a Cell Line HFH-T2, Producing Viral Particles, from Primary Human Fetal Hepatocytes Infected with Hepatitis B Virus

  • Shim, Jae-Kyoung;Kim, Dong-Wook;Chung, Tae-Ho;Kim, June-Ki;Suh, Jeong-Ill;Park, Chun;Lee, Young-Choon;Chung, Tae-Wha;Song, Eun-Young;Kim, Cheorl-Ho
    • Journal of Microbiology and Biotechnology
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    • v.11 no.2
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    • pp.186-192
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    • 2001
  • A primary culture of human fetal hepatocytes was obtained through a therapeutic abortion process at 26 weeks of gestation period. More than $10^8$ cells were seeded on a plastic plate. These hepatocytes were infected with hepatitis B virus (HBV). The HBV was purified from serum of one chronic HBV carrier. Transformed hepatocytes were subcultured in a 10% FBS-supplemented medium. The morphology of the transformed cell was epithelial-like. The cells from the first pass showed signs of early proliferation and had a latent period of more than 3 months after 6-7 passages. After the rest period, the transformed cell proliferated actively and they were subcultured every three days. Transformed hepatocytes were characterized by detection of the HBV transcript by RT-PCR. The secretion of virions from transformed cells was investigated by PCR with the cell medium. Two types of virions secreted into the culture medium were examined by using the transmission electron microscope. Another approach to study the secretion of virions in to culture medium was carried out with HBV antibody. HBsAg was detected in the culture medium of transformed cells using ELISA and Western blot analyses. These data suggested that the human fetal hepatocyte cell line has been established by infection of HBV, in which this cell line secreted viral particles into the culture medium.

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Comparison of applicability of HVAC and air cleaners in a subway station platform against airborne infection of SARS-CoV-2 (지하역사 승강장에서 코로나19 바이러스 공기감염 대응 공기조화기와 공기청정기의 적용 효과 비교)

  • Lee, Gunhee;Kim, Sang Bok;Park, Inyong;Hong, Kee Jung;Lee, Yeawan;Kim, Hak-Joon;Kim, Yong-Jin;Han, Bangwoo
    • Particle and aerosol research
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    • v.18 no.3
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    • pp.51-59
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    • 2022
  • In this work, virion concentration and its dose changes by HVAC and air cleaners were estimated in a subway station platform to control airborne infection of SARS-CoV-2. Collection efficiencies with particle size were measured for the air filter equipped in a HVAC in one subway station in Daejeon. Indoor PM2.5 changes according to outdoor PM2.5 with time were also measured to estimate air infiltration rate in the subway station platform. When infected persons generate virions by 104, 105, 106, 3 × 106 and 5 × 106 h-1 in a 2,400 m3 volume platform, the concentration and dose were estimated as 9, 92, 275 and 458 virions/m3 and 4, 43, 130 and 217 virions after 1 hour exposure, respectively. The concentration and dose were reduced by 70%, and 64%, respectively by operations of both HVAC (with a flow rate of 16,000 m3/h, MERV 11) and ten air cleaners(with total CADR 10,740 m3/h) compared to those without operation of both HVAC and air cleaners. However, virion dose in the platform was estimated to be too low at the general conditions due to a large space, a high air infiltration (3 h-1) and a short residence time (usually < 10 mins) in the platform irrespective of the operations of HVAC or air cleaners. HVAC with filters and air cleaners would be more necessary in the concourse or shopping areas in the subway stations to reduce the infection dose from a few hundred to several tens virions in a hour.

Study or The Genetic Characteration of Herpes Simplex Virus (Herpes simplex 바이러스의 유전학적 특성에 관한 연구)

  • Kang, Bong-Joo;Choi, Whan-Soo;Choi, Sun-Mi;Shin, Hyun-Kyoo;Cho, Dong-Wuk;Park, Kap-Joo
    • Korean Journal of Oriental Medicine
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    • v.1 no.1
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    • pp.477-493
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    • 1995
  • In order to facilitate the molecular characterization of the Herpes simplex Virus types 1 and types 2 genome DNAs, a gene library of cloned restriction frtgments have been produced. The Vero cells were infected with HSV-1 and HSV-2. 48 hours after infection, the infected cells Ivere Iysed, and multinucleated giant cells were observed approximately at seventy-two hours postinfection. The multiplication of HSV-1 and HSV-2 was observed in Vero cells using electromicroscopy. The nucleocapsids in nuclei were obseryed, and the assembled virions were budded out through the vacuole, and the virions were released from the cells. HSV-1 and HSV-2 was analyzed by digestion of their genome DANs with restriction ensymes. HSV-1 and HSV-2 genome DNAs were digested with BarnHI, Bgfl respectively. The BarnHI rlestriction fragments of HSV-1 and HSV-2 genome DNAs were twenty-seven fragments and thair molecular sizes were ranging $0.70{\sim}15.08$, $4.4{\sim}31.0$ tilobases. The BglII restriction fragments of HSV-1 and HSV-2 genome DNAs were sixteen, eighteen fragments and thair molecular sizes were ranging $4.8{\sim}30.0$, $1.2{\sim}25.0$ kilobases. And then BglII restriction frgments were cloned in Escherichia coli(E.coil) using the plasmid vector pBacPAK9.

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Modification of Turnip yellow mosaic virus coat protein and its effect on virion assembly

  • Shin, Hyun-Il;Chae, Kwang-Hee;Cho, Tae-Ju
    • BMB Reports
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    • v.46 no.10
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    • pp.495-500
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    • 2013
  • Turnip yellow mosaic virus (TYMV) is a positive strand RNA virus. We have modified TYMV coat protein (CP) by inserting a c-Myc epitope peptide at the N- or C-terminus of the CP, and have examined its effect on assembly. We introduced the recombinant CP constructs into Nicotiana benthamiana leaves by agroinfiltration. Examination of the leaf extracts by agarose gel electrophoresis and Western blot analysis showed that the CP modified at the N-terminus produced a band co-migrating with wild-type virions. With C-terminal modification, however, the detected bands moved faster than the wild-type virions. To further examine the effect, TYMV constructs producing the modified CPs were prepared. With N-terminal modification, viral RNAs were protected from RNase A. In contrast, the viral RNAs were not protected with C-terminal modification. Overall, the results suggest that virion assembly and RNA packaging occur properly when the N-terminus of CP is modified, but not when the C-terminus is modified.

Protection of Specific-pathogen-free (Spf) Foals from Severe Equine Herpesvirus Type-1 (Ehv-1) Infection Following Immunization with Non-infectious L-particles

  • Mohd Lila Mohd-Azmi;John Gibson;Frazer Rixon;Lauchlan, John-Mc;Field, Hugh-John
    • Journal of Microbiology
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    • v.40 no.3
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    • pp.183-192
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    • 2002
  • Cells infected With equine herpesvirus type-1 (EHV-1) Produced both infectious and non-infectious Virus-related particles. Compared to the whole virion, non-infectious particles termed L-particles were deter-mined to lack 150 kDa protein, commonly known as nucleocapsid protein. The potential of L-particles to induce immune responses was studied in mice and foals. Intranasal immunization with L-particles or whole virions induced poor IgG antibody responses in mice. Interestingly, despite the poor antibody response, the conferred immunity protected the host from challenge infections. This was indicated by a significant reduction in virus titers in line with recovery towards normal body weight. Subsequently, the test on the usefulness of L-particles as immunizing agents was extended to foals. Immunization of specific-pathogen-free (SPF) foals resulted in similar results. As determined by a complement-fixing-antibody test (CFT), foals seroconverted when they were immunized either with inactivated L-particles or whole virions via intramuscular (i.m.) injections. The presence of the antibody correlated with the degree of protection. Beyond day 1 post challenge infection (p.i.), there was no virus shedding in the nasal mucus of foals immunized with whole EHV-1 virions. Virus shedding was observed in foals Immunized with L-particles but limited to days 6 to 8 p.i. only. In contrast, extended vim shedding was observed in non-immunized foals and it was well beyond day 14 p.i. Viremia was not detected for more than four days except in non-immunized foals. Immunization in mice via intranasal (i.n.) conferred good protection. However, compared to the i.n. route, a greater degree of protection was obtained in foals following immunization via i.m. route. Despite variation in the degree of protection due to different routes of immunization in the two animal species, our results have established significant evidence that immunization with L-particles confers protection in the natural host. It is suggested that non-infectious L-particles should be used as immunizing agents for vaccination of horses against EHV-1 infection.

3' end of putative sequences of the packaging signal in moloney-murine leukemia virus (Moloney murine Leukemia Virus에서 포장신호의 가능한 3' 끝의 염기서열)

  • 박종상
    • Korean Journal of Microbiology
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    • v.26 no.2
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    • pp.101-105
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    • 1988
  • 6M-MuLV mutants containing deldtions around the putative packaging signal were constructed by using recombinant DNA technique and transfected into NIH/3T3 cell. 2 of 6 mutants can not be packaged into virions even in the presence of the wild type helper virus. The boundary between the packagible and the non-packagible genome is located around Pvu I site, 421 nucleotide downstream from the 5' end of M-MuLV genome. 10 base pair inverted repeat sequence (GAGUCCAAAA) which can make stem structure around Pvu Isite could be the putative packaging signal.

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Infection and Pathogenesis Mechanisms of Marek's Disease Virus (마렉병 바이러스 감염과 병원성 발현 기전)

  • Jang, H.K.;Park, Y.M.;Cha, S.Y.;Park, J.B.
    • Korean Journal of Poultry Science
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    • v.35 no.1
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    • pp.39-55
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    • 2008
  • Like the other herpesviruses, the virion of MDV consists of an envelope, which surrounds an amorphous tegument. Within the tegument, and icosahedral capsid encloses a linear double-stranded DNA core. Although the genome structure of MDV indicates that it is an ${\alpha}-herpesvirus$ like herpes simplex and varicella-zoster viruses, biological properties indicate MDV is more akin to the ${\gamma}-herpesvirus$ group, which includes Epstein-Barr and Kaposi's sarcoma herpesviruses. These herpesviruses replicate lytically in lymphocytes, epithelial and fibroblastic cells, and persist in lymphoblastoid cells. MDV has a complex life cycle and uses two means of replication, productive and non-productive, to exist and propagate. The method of reproduction changes according to a defined pattern depending on changes in virus-cell interactions at different stages of the disease, and in different tissues. Productive (lytic) interactions involve active invasion and take-over of the host cell, resulting in the production of infectious progeny virions. However, some herpesviruses, including MDV, can also establish a non-productive (abortive) infection in certain cell types, resulting in production of cell-associated progeny virus. Non-productive interactions represent persistent infection, in which the viral genome is present but gene expression is limited, there is no structural or regulatory gene translation, no replication, no release of progeny virions and no cell death. Reactivation of the virus is rare, and usually the infectious virus can be re-isolated only after cultivation in vitro. MDV establishes latency in lymphoid cells, some of which are subsequently transformed. In this review article, recent knowledges of the pathogenesis mechanisms followed by MDV infection to sensitive cells and chickens are discussed precisely.