• 제목/요약/키워드: RTase activity

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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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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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Arrest of Cell Growth by Inhibition of Endogenous Reverse Transcription Activity in Cancer and Somatic Cell Lines (사람의 암세포주 및 정상세포주에서 역전사 효소의 억제에 의한 세포 성장의 제한)

  • Mi-Jeong Kim;Sung-Ho Lee;Jong-Kuen Park;Byeong-Gyun Jeon
    • Journal of Life Science
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    • v.34 no.6
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    • pp.365-376
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    • 2024
  • The present study assessed the cytotoxic effects on cell growth and senescence in human cancer (A-549, AGS, HCT-116, MDA-MB-231, and U 87-MG) and normal (MRC-5 and mesenchymal stem cells) cell lines treated with efavirenz (EFA), an inhibitor of non-nucleoside reverse transcriptase (RTase). Following EFA treatment, the half-maximal inhibitory concentration (IC50) values were approximately 15 µM, and the IC50 value was significantly (p<0.05) lower in the cancer cell lines, compared to normal cell lines. After determining the IC50 values against EFA, each cell line was treated with 15 µM EFA for up to one week. Significant (p<0.05) decreases in endogenous RTase and telomerase activity were observed in the cancer cell lines. RTase and telomerase activity were absent or detected at very low levels in both EFA-untreated and treated MRC-5 and MSC normal cells. The cell doubling time (CDT) was also significantly (p<0.05) prolonged by the decreased cell growth rate in the EFA-treated cancer cell lines compared to the untreated cell lines. Furthermore, EFA-treated cancer cells displayed a high number of cells with a high intensity of senescence-associated ß-galactosidase activity (SA-ß-gal activity), compared to the untreated cells. The present study showed that inhibition of RTase activity induces cellular senescence and arrests cell growth in human cancer cell lines; however, normal cell lines showed greater tolerance against EFA. RTase treatment could offer optional chemotherapy for cancer treatment in human cancer cell lines with high RTase activity.

Identification of anti-HIV and anti-Reverse Transcriptase activity from Tetracera scandens

  • Kwon, Hyeok-Sang;Park, Jung-Ae;Kim, Joo-Hwan;You, Ji-Chang
    • BMB Reports
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    • v.45 no.3
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    • pp.165-170
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    • 2012
  • We report here that an ethanol extract of Tetracera scandens, a Vietnamese medicinal plant, has anti-HIV activity and possesses strong inhibitory activity against HIV-1 reverse transcriptase (RTase). Using a MT-4 cell-based assay, we found that the T. scandens extract inhibited effectively HIV virus replication with an $IC_{50}$ value in the range of 2.0-2.5 ${\mu}g$/ml while the cellular toxicity value (CC50) was more than 40-50 ${\mu}g$/ml concentration, thus yielding a minimum specificity index of 20-fold. Moreover, the anti-HIV efficacy of the T. scandens extract was determined to be due, in part, to its potent inhibitory activity against HIV-1 RTase activity in vitro. The inhibitory activity against the RTase was further confirmed by probing viral cDNA production, an intermediate of viral reverse transcription, in virus-infected cells using quantitative DNA-PCR analysis. Thus, these results suggest that T. scandens can be a useful source for the isolation and development of new anti-HIV-1 inhibitor(s).

Biological Activities of Phloroglucinol Derivatives from Eucalyptus Spp.

  • Singh, Inder Pal;Etoh, Hideo
    • Natural Product Sciences
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    • v.3 no.1
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    • pp.1-7
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    • 1997
  • Eucalyptus is a rich source of biologically active compounds. Among these, phloroglucinol compounds such as sideroxylonals, macrocarpals, euglobals, and robustadials are unique to Eucalyptus species. Sideroxylonal A is a very potent attachment-inhibitor. Macrocarpals show very strong antibacterial activity against gram positive bacteria. Macrocarpals also show HIV-RTase inhibitory activity. Euglobals are potent inhibitors of Epstein-Barr virus activation and are developed as skin and antitumor agents. They also show granulation inhibitory activity. In this review we aim to remove the existing confusion in literature on macrocarpals and discuss the biological activities and structure-activity relationships of phloroglucinol compounds.

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