• Title/Summary/Keyword: Transcriptase

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CDDP induces conformational changes in BTV ds RNA rather than forming protein-protein and/or protein-RNA crosslink (cis-Diamminedichloroplatinum(II) (CDDP)에 의한 불루텅 바이러스 이중가닥 RNA의 구조변화)

  • Yang, Jai-Myung
    • Applied Biological Chemistry
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    • v.34 no.2
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    • pp.86-93
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    • 1991
  • cis-Diamminedichloroplatinum(II)(CDDP), an antitumor drug, did not generate crosslink between bluetongue virus (BTV) capsid protein at moderate concentration. Cesium chloride density gradient centrifugation study revealed that protein-RNA crosslink was not detectable in CDDP treated BTV. CDDP treated BTV ds RNA showed remarkable change in the migration pattern in polyacrylamide gel electrophoresis. These results suggest that the reduction of BTV core associated transcriptase activity is most likely by the CDDP adduction to the genomic ds RNA rather than by the protein-RNA crosslink and/or protein-protein cross-link.

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CAT 유전자를 지닌 HIV-1을 이용한 시험관내 항 AIDS 약물의 약효 검색

  • 성영철
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1993.04a
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    • pp.80-80
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    • 1993
  • (목적) 본 연구에서 사용된 바이러스는 HIV-1 nef유전자가 일부 삭제되고 대신 Chloramphenicol acetyltransferase(CAT)가 pSVCAT recombinant 바이러스다. 이러한 recombinant 바이러스를 사용하는 이유는 첫째, CAT activity가 매우 민감하므로 바이러스의 복제억제 정도를 정확하게 측정 할 수 있고 둘째, simian immunodeficiency virus(SIV)의 경우 nef 유전가 in vivo에서는 바이러스의 복제에 필수적이므로 HIV가 SIV와 유사한 것으로 미루어 본 연구에서 사용되는 recombinant SVCAT 바이러스가 안전한 것으로 고려되기 때문이다. (방법) 특히 화합물이 HIV-1의 복제에 얼마나 영향이 있는가는 1) 어느정 도의 virus inoculm을 넣었는지 2) 사용하는 cell line 3) 사용한 cell line의 infection kinetics 4) 실험의 지속기간 5) 테스트하는 assay의 sensitivity에 의존한다. 따라서 $10^{5}$ cell의 H9과 sup T1을 24 well plate에 넣고 sup T1 cell line의 경우 3일 후 항 화합물에 의한 syncytia 형성 및 CAT activity의 억제정도를 현재 AIDS drug으로 쓰이고 있는 Zidovudine을 control로 비교 관찰하였다. H9 cell line의 경우 3일 간격으로 media의 3/4을 fresh media로 바꾸어 주고 9일 후 CAT assay를 하였다. 이러한 assay에서 activity를 보이는 화합물을 reverse transcriptase와 P24 ELISA assay를 재확인하였다.다.

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Capsid Protein Gene Sequence Analysis and Development of Diagnostic Method by RT-PCR of Barley Yellow Mosaic Virus

  • Lee, Kui-Jae;So, In-Young
    • Plant Resources
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    • v.2 no.2
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    • pp.69-74
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    • 1999
  • A rapid and sensitive assay for specific detection and identification of barley yellow mosaic virus(BaYMV) was set up using the reverse transcriptase polymerase chain reaction(RT-PCR). A couple of primers was select to discriminate the viruses. PCR fragments of BaYMV(ca.0.9 kb) were obtained by using the method designed for BaYMV capsid protein. RT-PCR fragments were cloned with vector pT7 Blue and the resulting clones were sequenced. Capsid protein of BaYMV consisted of 297 amino acids and 891 nucleotides. The capsid protein sequence of BaYMV showed that 98% of nucleotides and 99% of amino acids homology.

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사람 T-임파구의 HPRT유전자에서 방사선 및 환경독성물질에 노출에 의한 돌연변이 빈도의 변화

  • 윤병수;조명행;이영순;김인규;이강석
    • Proceedings of the Korean Nuclear Society Conference
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    • 1996.05d
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    • pp.40-45
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    • 1996
  • 사람 T-임파구에서 방사선 독성물질인 감마선과 화학적 독성물질인 PCP(pentachlorophenol)의 돌연변이 효과를 hprt (hypoxanthine phosphoribosyl transferase) 유전자의 돌연변이빈도로써 측정하였다. 감마선은 $^{137}$Cs원을 사용하여, 0 - 300 rads의 양으로 세포의 초기배양 시기에 조사하였으며, PCP는 역시 세포의 초기배양시 최종농도 0-100 ppm으로 24시간 투여 하였다. 양 돌연변이원은 hprt유전자를 돌연변이 시키어, 300rads의 조사는 대조군에 비하여 약 7.5배의 돌연변이빈도의 증가를 나타내었고, PCP 500 ppm의 처리는 대조군에 비하여 약 5배의 돌연변이빈도 증가를 나타내었다. 본 실험에 사용된 상이한 두 돌연변이원은 모두 비교적 정확한 용량-반응 관계를 보였으나, 환경독성물질들의 혼합효과에서 돌연변이원을 정성하기위하여 개선된 T-cell hprt clonal assay, 즉 reverse transcriptase/polymerase chain reaction 및 direct sequencing에 의한 mutational spectrum의 적용이 요구되었다.

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Cloning and Expression of a cDNA AAPT3 Encoding Aminoalcoholphosphotransferase Isoform from Chinese Cabbage

  • Kim, Kwang-Soo;Park, Jong-Ho;Cho, Sung-Ho
    • Animal cells and systems
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    • v.8 no.2
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    • pp.105-109
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    • 2004
  • Aminoalcoholphosphotransferase catalyzes the synthesis of phosphatidylcholine and phosphatidylethanolamine from diacylglycerol plus a CDP-aminoalcohol such as CDP-choline or CDP-ethanolamine. Previously we suggested the presence of possible isoforms of this enzyme from Chinese cabbage roots and now report the cDNA cloning and expression analysis of AAPT3 encoding a third isoform of aminoalcoholphosphotransferase (AAPT3). AAPT3 contains an open reading frame of 1,176 bp coding for a protein of 392 amino acids. It shares 96 and 95% identity with Chinese cabbage AAPT1 and AAPT2, respectively, at the deduced amino acid level. The results from reverse transcriptase-polymerase chain reaction analysis indicate that expression of AAPT3 is up-regulated by low temperature as well as AAPT1 and AAPT2.

Vitellogenin mRNA Induction in Male African Clawed Frog Treated with di-2-ethylhexyl Phthalate (Di-2-ethylhexyl phthalate 처리 남아프리카산발톱개구리에서의 vitellogenin 발현)

  • 박응로;이철우;류지성;남성숙;전성환;나진균;최덕일;박광식
    • Environmental Analysis Health and Toxicology
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    • v.16 no.1
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    • pp.29-34
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    • 2001
  • The estrogenic potency of di -2-ethylhexyl phthalate (DEHP) using reverse transcriptase-PCR respouse of liver vitellogenin mRNA in male African clawed frog (Xenopus laevis) was studied. Male frogs were injected with DEHP at dose of 300$\mu\textrm{g}$/kg and 300 mg/kg body weight through the dorsal lymph sac. After 4 days, using suitable pair of RT-PCR primers, vitellogenin mRNA induction in the liver was measured and DEHP showed vitellogenin mRNA induction in only the group treated with 300 mg/kg. Any significant histological abnormalities by the exposure of DEHP was not shown in both testis and liver.

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Isolation and Characterization of Cinnamoyl CoA Reductase Gene from Codonopsis lanceolata

  • Kim Ha-Na;In Jun-Gyo;Lee Eun-Kyung;Yoon Jae-Ho;Lee Mee-Sook;Bae Hyeun-Jong;Yang Deok-Chun
    • Plant Resources
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    • v.8 no.3
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    • pp.181-187
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    • 2005
  • A cinnamoyl CoA reductase (CCR) cDNA (ClCCR) was isolated from tabroot mRNAs of Codonopsis lanceolata by cDNA library construction, and its expression was investigated in relation to abiotic stresses. The ClCCR is 1008 bp in length with an open reading frame (ORF) of 336 amino acids. The deduced amino acid sequence was showed high similarity with cinnamoyl-CoA reductases of P. tremuloides (AAF43141) 87%, F.${\times}$aranassa (AAP46143) 83%, L. album (CAD29427) 80%, E. gunnii (CAA66063) 72%, S. tuberosum (AAN71761) 83%. Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis was revealed that the ClCCR expression was regulated by abiotic stresses.

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Herbicide Resistant Turfgrass(Zoysia japonica cv. 'Zenith') Plants by Particle bombardment-mediated Transformation

  • Lim Sun-Hyung;Kang Byung-Chorl;Shin Hong-Kyun
    • Asian Journal of Turfgrass Science
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    • v.18 no.4
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    • pp.211-219
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    • 2004
  • Transgenic zoysiagrass (Zoysia japonica cv. Zenith) plants have been obtained by particle bombardment of embryogenic callus with the plasmid pSMABuba, which contains hygromycin resistance (hpt) and bialaphos resistance (bar) genes. Parameters on DNA delivery efficiency of the particle bombardment were partially optimized using transient expression assay of a chimeric $\beta-glucuronidase$(gusA) gene driven by the CaMV 35S promoter. Stably transfarmed zoysiagrass plants were recovered with a selection scheme using hygromycin. Transgenic zoysiagrass plants were confirmed by PCR analysis with specific primer for bar gene. Expression of the transgene in transformed zoysiagrass plants was demonstrated by Reverse transcriptase (RT)-PCR analysis. All the tested transgenic plants showed herbicide BastaR resistance at the field application rate of $0.1\%-0.3\%$.

Investigation of gene expression of GX-12, a new DNA vaccine for HIV infection, in reproductive organs in SD rats

  • Kang, Kyung-Koo;Park, Min-Seul;Ahn, Kook-Jun;Baik, Dae-Hyun;Sohn, Yong-Sung;Lee, Dong-Sup;Park, Jae-Hun;Ahn, Byoung-Ok;Kim, Won-Bae
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2002.11b
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    • pp.201-201
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    • 2002
  • GX-12 is a naked DNA vaccine developed by Dong-A pharmaceutical company for the treatment of mv infection. GX-12 consists of four separate plasmids. This study was performed to evaluate the biodistribution and expression of GX-12 mRNA in gonadal tissues, and to investigate the histopathological changes in rats after repeated intramuscular injection.(omitted)

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Effects of Wogonin, a Plant Flavone from Scutellaria Radix, on Skin Inflammation:In Vivo Regulation of Inflammation-associated Gene Expression

  • Chi, Yeon-Sook;Lim, Hyun;Park, Hae-Il;Kim, Hyun-Pyo
    • Proceedings of the PSK Conference
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    • 2003.10b
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    • pp.155.3-156
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    • 2003
  • Flavonoids from plant origin show anti-inflammatory activity in vitro and in vivo. In addition to inhibition of inflammation-associated enzymes such as cyclooxygenases and lipoxygenases, they have been found to regulate the expression of inflammation-associated proteins from in vitro experiments. In order to prove in vivo behavior and the potential for beneficial use against inflammatory skin disorders, the effect of wogonin (5,7-dihydroxy-8-methoxyflavone) on in vivo expression of several inflammation-associated genes was examined in the intact as well as in the inflamed mouse skin by reverse transcriptase-polymerase chain reaction analysis. (omitted)

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