• Title/Summary/Keyword: virus replication

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Cloning, Expression and Purification of HIV-1 Reverse Transcriptase

  • Goo, Jae-Hwan;Park, Kwan-Yong
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1995.04a
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    • pp.76-76
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    • 1995
  • Virus-encoded HIV-1 reverse transcriptase (RTase) is one of the major targets for the development of drugs for HIV-1 since it is an essential enzyme-for the replication cycle of HIV-1. We cloned the entire reverse trancriptase gene into an inducible expression vector with tac promotor= RTase was stably overexpressed and induced by IPTG and the highly-expressed RTase was purified partially by use of DEAE cellulose and Mono Q column. The partially purified enzyme (663kDa, 51kDa) as exhibited by SDS-PAGE showed the high specific activity (16,570U/mg) when the assay for the RTase activity was carried out using $^3$H-dTTP and poly(rA): oligo(dT)12-18 as the substrate.

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Analysis of In Vivo Interaction of HCV NS3 Protein and Specific RNA Aptamer with Yeast Three-Hybrid System

  • HWANG BYOUNGHOON;LEE SEONG-WOOK
    • Journal of Microbiology and Biotechnology
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    • v.15 no.3
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    • pp.660-664
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    • 2005
  • We have previously isolated specific RNA aptamers with high affinity against the helicase domain of hepatitis C virus (HCV) nonstructural protein 3 (NS3). The RNA aptamers competitively and efficiently inhibited the helicase activity, partially impeding HCV replicon replication in human hepatocarcinoma cells. In this study, the RNA aptamers were tested for binding to the HCV NS3 proteins in eukaryotic cells, using a yeast three-hybrid system. The aptamers were then recognized by the HCV NS3 proteins when expressed in the cells, while the antisense sequences of the aptamers were not. These results suggest that the in vitro selected RNA aptamers can also specifically bind to the target proteins in vivo. Consequently, they could be potentially utilized as anti-HCV lead compounds.

항바이러스제가 단백질의 구조적 거동에 미치는 영향에 대한 유한요소법 기반 분석

  • Yun, Gi-Seok;Kim, Jae-Hun
    • Proceeding of EDISON Challenge
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    • 2015.03a
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    • pp.212-216
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    • 2015
  • Oseltamivir, also known as Tamifu, is an inhibitor of neuraminidase protein which plays an essential role in proliferation and replication of influenza virus. Binding to the active site of neuraminidase, the oseltamivir prevents the protein from enzyme reaction. Conformational change of the protein(neuraminidase) should be accompanied by the enzyme reaction, but the drug inhibits the protein to deform. In this study, we examine the influence of oseltamivir on protein's conformational change in the structural and mechanical point of view. Finite element analysis of the protein can be an useful approach to investigate the influence of oseltamivir on the deformation of a protein. We suggest the finite element based protein model, and then perform the linear static analysis with the displacement loading condition based on the first two largest motion which can be obtained from the normal mode analysis. The results show that it takes more energy to change shape of the protein with an oseltamivir attached than the protein without an oseltamivir.

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Antiviral Activity of Papaverine and Nucleoside Analogs on the Human Cytomegalovirus Infection (Human Cytomegalovirus 감염에 대한 파파베린과 뉴클레오사이드 유사체의 항바이러스 효과)

  • ;Albrecht, T.
    • Korean Journal of Microbiology
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    • v.29 no.1
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    • pp.25-33
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    • 1991
  • Antiviral activities of papaverine and nucleoside analogs, 9-[(1,3-dihydroxy-2-propoxy)methyl]guanine (DHPG) and acyclovir, against human cytomegalovirus (HCMV) infection were compared in vitro. Papaverine and DHPG were effective in reducing infectious HCMV yields with $ED_{50}{\s}$ (effective dose 50: the concentraion at which 50% of virus yields was obtained) of approximately 1.02 and $0.45{\mu}{\M}$, respectively; while acyclovir was less effective with an $ED_{50}$ of about $10.4{\mu}{\M}$The relative cytotoxicity of these drugs was evaluated under the same conditions used to measure infectious HCMV yields. Papaverine and DHPG demonstrated little cellular toxicity as measured by their effect on the viability of confluent cells at concentrations in the range of those demonstrating potent inhibition of HCMV replication. Similarly, protein synthesis was largely unaffected by these drugs in stationary mock-infected cells as measured by the incorporation of isotopically labelled amino acids. In contrast, cellular DNA synthesis was invariably reduced in the presence of either drug. HCMV-specific DNA synthesis was also strongly inhibited by papaverine and DHPG.

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Cytomegalovirus-associated esophageal ulcer in an immunocompetent infant: When should ganciclovir be administered?

  • Jang, Hyo-Jeong;Kim, Ae Suk;Hwang, Jin-Bok
    • Clinical and Experimental Pediatrics
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    • v.55 no.12
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    • pp.491-493
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    • 2012
  • Cytomegalovirus (CMV)-associated esophageal ulcer is rare in immunocompetent infants. The presence of inclusion bodies and immunohistochemical staining for CMV in biopsy specimens obtained during esophagogastroduodenoscopy (EGD) indicate that such ulcers occur because of CMV infection. A 7-week-old female infant who experienced frequent vomiting and feeding intolerance was diagnosed with a massive CMV-associated ulcer in the distal esophagus. The ulcer improved after conservative treatment using proton-pump inhibitors; however, ganciclovir was not administered. In a follow-up EGD biopsy specimen, no CMV inclusion bodies were present, and immunohistochemical staining results for this virus were negative. The presence of CMV inclusion bodies indicates active viral replication. If persistent inclusion bodies or positive immunohistochemical staining for CMV is observed in follow-up biopsy specimens, ganciclovir may be used to treat CMV-associated esophageal ulcers.

Impact of SV40 T antigen on two multiple fission microalgae species Scenedesmus quadricauda and Chlorella vulgaris

  • Gomaa, Ahmed E.;Yang, Seung Hwan
    • International journal of advanced smart convergence
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    • v.7 no.1
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    • pp.48-63
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    • 2018
  • The combination of Simian Virus40 (SV40)'s large T antigen with its replication origin is commonly used in molecular studies to enhance the expression of heterogeneous genes through multiplying the plasmid copy number. There are no reports related to the impact of the SV40 T antigen on plant, multiple fissional, cell-type. This study explores the response of two multiple-fission microalgal cells, Scenedesmus quadricauda and Chlorella vulgaris, to the expression of the T-antigen, with aim of applying SV40 T-antigen to increase the expression efficiency of foreign genes in the two species. Different levels of low-expression have been constructed to control the expression of SV40 T antigen using three heterogenous promoters (NOS, CaMV35S, and CMV). Chlorella cultures showed slowdown in the growth rate for samples harboring the T antigen under the control of CaMV35S and CMV promoters, unlike Scenedesmus cultures which showed no significant difference between samples and could have silenced the expression.

Construction of an expression vector with SV40 DNA in a mammalian cell (SV 40 DNA를 이용한 포유동물의 유전자 운반체 개발)

  • 정민혜;김상해;전희숙;노현모
    • Korean Journal of Microbiology
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    • v.25 no.3
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    • pp.165-172
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    • 1987
  • An expression vector in a mammalian cell was constructed using the origin of replication (OR) and the promoters of SV40. The plasmid pSVOE was constructed by inserting SV40 DNA fragment (1, 118bp) containing SV40 OR and promoters into pBR322-1, and then a multiple cloning sequence was inserted at the immediate downstream of the late promoter of SV40 in the pSVOE vector. The plasmid was named pSVML. As a selection marker, thymidine kinase gene of herpes simplex virus with its promoter was inserted into EcoRI site of pSVML and the recombinant was named pSVML-TKp. To test the expression capacity of foreigen gene inserted at the multiple cloning site of pSVML, the thymidine kinase gene without its own promoter was inserted at the BamHI site of pSVML. The recombinant was named pSVML-TK. These plasmids, pSVML-TKp and pSVML-TK, were transfected into COS cells with calcium phosphate precipitation method. The thymidine kinase activity was significantly increased in both transfected cells.

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Development of Drug Candidates based on Natural Products Against COVID-19 (천연식물자원 활용 코로나19 억제 치료제 개발)

  • Se Chan Kang
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2021.04a
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    • pp.3-3
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    • 2021
  • The ongoing global pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has not only influenced over 1.26 billion people but also caused 2.77 million deaths worldwide (as of March 28, 2021). The vaccination could be the most efficient strategy to prevent SARS-CoV-2 infection. However, the continuous emergence of novel variants such as VUI-202012/01 (United Kingdom) and 501.V2 (South Africa) raises huge concerns about the effectiveness of the vaccine designed to target the original virus strain. Since ancient times regardless of the East and West, the plants which refered in this presentation have been consumed not only as food but also as a natural medicine to treat diverse diseases including infectious diseases. Importantly, these plants contain secondary metabolites that display antiviral activity involved in the inhibition of viral adsorption, penetration, and replication. Also, plant-derived natural medicines are expected to have a wider range of efficacy and fewer side effects than synthetic medicine, discovering novel plant-based viral agents would be a promising strategy to fight against SARS-CoV-2.

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Poxvirus under the eyes of electron microscope

  • Jaekyung Hyun
    • Applied Microscopy
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    • v.52
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    • pp.11.1-11.9
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    • 2022
  • Zoonotic poxvirus infections pose significant threat to human health as we have witnessed recent spread of monkeypox. Therefore, insights into molecular mechanism behind poxvirus replication cycle are needed for the development of efficient antiviral strategies. Virion assembly is one of the key steps that determine the fate of replicating poxviruses. However, in-depth understanding of poxvirus assembly is challenging due to the complex nature of multi-step morphogenesis and heterogeneous virion structures. Despite these challenges, decades of research have revealed virion morphologies at various maturation stages, critical protein components and interactions with host cell compartments. Transmission electron microscopy has been employed as an indispensable tool for the examination of virion morphology, and more recently for the structure determination of protein complexes. In this review, we describe some of the major findings in poxvirus morphogenesis and the contributions of continuously advancing electron microscopy techniques.