• Title/Summary/Keyword: DNA합성

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HIV-1 RT (reverse transcriptase) 저해제에 대한 내성 발현 기전

  • 임광진
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1995.10a
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    • pp.67-69
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    • 1995
  • reverse transcription은 AIDS를 일으킨다고 알려진 바이러스인 HIV-1의 번식에는 필수적이나 인체 세포에는 필수적이 아니기에 이 단계를 표적으로 하는 AIDS 치료제가 우선적으로 개발되었다. 그 단계에 필요한 효소가 바이러스에 의해 만들어진 RT이며 이 효소의 작용을 저해하는 nucleoside 유도체들인 AZT, DDC, DDI 들이 현재 AIDS 환자의 치료에 사용되고 있다. 이들 nucleoside 유도체들은 세포안으로 들어가 triphosphate 형태로 변화된 후 dNTP와 상경적으로 경쟁하며 합성 중인 바이러스의 DNA에 들어가 DNA의 합성을 정지시켜 바이러스의 증식을 억제한다. 그러나, 이들 nucleoside 유도체들은 치료용량에서 심한 독성을 나타낼 뿐만 아니라 장기 투여시 내성을 나타내는 바이러스가 생겨나 AIDS의 치료를 불가능하게 하고 있다.

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1984~1985 미국 100대 발명, 발견

  • Korean Federation of Science and Technology Societies
    • The Science & Technology
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    • v.19 no.2 s.201
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    • pp.79-84
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    • 1986
  • - 환경내의 유전물질 빈도를 모니터 하기 위한 DNA 탐지장치 - RRV1 및 RRV2 로보트 - 기능적 특성을 예견하기 위한 비천연 단백질의 설계와 합성 - 고분해능의 주사 이온 미세탐침 - 에크타켐 DT60 혈액분석기 - 뇌일혈 치료시스템 - 레일건 - 소프트스트립 - 고수준 핵폐기물 저장용 납-철-인산염 유리공정 - 단백질과 DNA의 합성 및 분석 - 물의 광전해용 전극 - 애버트 비루스용 고급휴지 - 3480 자기테이프 서브시스템

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Campylobacter jejuni 의 열충격 반응과 그유전자에 관한 연구

  • 김치경;임채일;이길재
    • Korean Journal of Microbiology
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    • v.30 no.3
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    • pp.232-238
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    • 1992
  • Canz~~j~lohuc;tc.~jurn i werc studied for their heat shock responses at several elevated temperatures and their heat shock genes were detected by the technique of Southern hybridization. (.. ,jc\ulcorneruni sy~>thesized the major heat shock proteins of hsp90. hsphh. and hsphO at 48$^{\circ}$C . ant1 their w~u.ival rates were maintained as the same level at optimal temperature. '1-hc heat shock genes in chromosome of C ,jc:jutii werc determined to be homologous to the heat shock genes or E. t,oli. by showing strong signals in Southern hybridization analysis using clnaK and groESL- as DNA probe But the restriction sites for thc fragmcnts including heat shock genes were different betueen E. c,oli and C ,jtjuni.

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Cloning of $\alpha$-Amylase Gene from Zea mays (옥수수 $\alpha$-amylase 유전자의 클로닝)

  • 김용욱;강신혜
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.38 no.3
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    • pp.275-282
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    • 1993
  • The objective of this study was to clone a partial fragment of $\alpha$-amylase from Korean maize. We designed and synthesized an oligonucleotide probe and two kinds of PCR primers based on cDNA conserved region of $\alpha$-amylase sequences from other plants. Total RNA from 3-day-old maize seedling was used as template for 1st strand cDNA synthesis and RNA-DNA hybrid was used as template for polymerase chain reaction(PCR). The product of PCR was about 0.5 kb long and inserted into pUC19. We named this recombinant plasmid as pZM$\alpha$'. The cloned fragment was certified by Southern blot analysis using labeled synthetic oligonucleotide as probe.

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The Effect of TGF-{\beta}_1 on Cellular Activity of Periodontal Ligament Cells activated by PDGF-BB (PDGF-BB에 의한 치주인대세포활성에 대한 TGF-{\beta}의 효과)

  • Baek, Sang-Churl;Park, Jin-Woo;Suh, Jo-Young
    • Journal of Periodontal and Implant Science
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    • v.32 no.3
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    • pp.457-473
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    • 2002
  • The purposes of this study is to evaluate the combination effects of TGF-${\beta}_1$ and PDGF-BB on the periodontal ligament cells to use as a regeneration promoting agent of periodontal tissue. Human periodontal ligament cells were prepared from the first premolar tooth extracted for the orthodontic treatment and were cultured in DMEM/100% FBS at the $37^{\circ}C$, 5% $CO_2$ incubator. Authors measured the DNA synthesis, total protein, collagen and noncollagenous protein synthesis according to the concentration of TGF-${\beta}_1$,(1,5ng/ml) and PDGF-BB (1,10 ng/ml) in combination. To explore further this delayed effect of TGF-${\beta}_1$, we preincubated human periodontal ligament cells with TGF-${\beta}_1$ for 4 or 24 hours before PDGF-BB stimulation. The results were as follows: The DNA synthetic activity was increased dose dependently by TGF-${\beta}_1$, PDGF-BB. The combination of TGF-${\beta}_1$ and PDGF-BB consistently enhanced the DNA synthetic activity to PDGF-BB alone. The ability of TGF-${\beta}_1$ to enhance DNA synthetic activity in PDGF-BB treated periodontal ligament cells was dose dependent. The maximum mitogenic effect was at the 5ng/ml of TGF-${\beta}_1$ and l0ng/ml of PDGF-BB. Preincubation of cell with TGF-${\beta}_1$ resulted in significantly greater response to PDGF-BB at all TGF-${\beta}_1$ concentration studied, and may be useful for clinical application in periodontal regenerative procedures. The total protein, collagen and noncollagen synthesis was increased dose pendently by TGF-${\beta}_1$, PDGF-BB. The % of collagen was slightly decreased according to the concentration of TGF-${\beta}_1$, PDGF-BB. The effect of TGF-${\beta}_1$, PDGF-BB were not specific for collagen synthesis since it also increased noncollagenous protein synthesis. This study demonstrates that PDGF-BB is major mitogens for human periodontal ligament cells in vitro, and supports a role for TGF-${\beta}_1$ as a regulation of the mitogenic and total protein formation to PDGF-BB in these cells.

Characterization of growth hormone-like sequence of loach, Misgurnus mizolepis (미꾸라지 성장 호르몬 염기 서열의 특성에 대하여)

  • Kim, Jin-Kyung;Song, Young-Hwan
    • Journal of fish pathology
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    • v.7 no.2
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    • pp.95-103
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    • 1994
  • We have prepared cDNA libray of loach. M. mizolepis in order to isolate cDNA clone of growth hormone gene. Total RNA was isolated from pituitary of loach, and then mRNA was further purified from total RNA by oligo (dT)-coupled magnetic beads. The purified mRNA was used as substrates to prepare cDNA. The resulting cDNA was ligated into the EcoRV/Smal site of pBlueKS+. The ligation mixture have transformed E. coli JM109 strain with electroporator to obtain high yield of transformation efficiency. All the transformants was screened with DIG-labeled Tilapia growth hormone gene by high density colony hybridization. After isolating 10 putative colonies showing the positive signals, secondary colony hybridization and southern hybridization could confirm it as true clones. The nucleotide sequence of one candidate, pCGHI, was compared with 312 bp DNA fragment used as DNA probe and show 52% relative homology to Tilapia growth hormone gene.

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Structural and Functional Aspects of DNA Polymerase (DNA Polymerase의 구조 및 기능 연구)

  • Kim, Young Tae
    • Journal of Life Science
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    • v.3 no.4
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    • pp.194-208
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    • 1993
  • DNA 복제시 중추적 단백질은 DNA 합성을 수행하는 DNA polymerase이다. 따라서 DNA polymerase의 구조 및 기능에 대한 연구는 DNA polymerase의 중합반응에 대한 기작을 비롯하여 교정 및 수선기능에 대한 정보를 얻게 함으로써 복잡한 DNA 복제 기적을 이해하는 첩경이 된다. Bacteriophage T7의 Gene 5 단백질은 T7 DNA polymerase로 Richardson group에 의해 처음으로 발견되었으며, E. coli의 12 KDa thioredoxin과 tight complex를 형성한다. T7 DNA polymerase의 클로닝은 분자생물학의 새로운 장을 열어준 중요한 의미를 지닌다 . 본 연구에서는 T7 DNA polymerase의 구조적, 기능적 특성을 파악하고 DNA 염기서열 분석에의 응용 및 DNA 염기서열 결정을 위한 새로운 전략 및 최근연구 동향에 대해 기술하였다.

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Effect of Solvent Fractions from Methanol Extract of Doenjang on Inhibition of Growth and DNA Synthesis of Human Cancer Cells. (인체 암세포 성장 및 DNA 합성 억제에 미치는 된장 분획물의 영향)

  • LIM Sun-Young;Rhee Sook-Hee;Park Kun-Young
    • Journal of Life Science
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    • v.15 no.5 s.72
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    • pp.685-691
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    • 2005
  • Growth and DNA synthesis inhibitory effects of doenjang methanol extract and its solvent fractions on AGS human gastric adenocarcinoma cells, Hep 3B human hepatocellular carcinoma cells, HT-29 human colon cancer cells and MG-63 human osteosarcoma cells were studied. The treatment of doenjang methanol extract ($ 200{\mu}g/ml $) with the AGS, Hep 3B, HT-29 and MG-63 cancer cells after 6 days of incubation inhibited the growth of cancer cells by $32\%$, $51\%$, $84\%$ and $33\%$, respectively. To separate active compounds of doenjang, doenjang methanol extract was fractionated with dichloromethane, ethylacetate, and buthanol. Among the solvent fractions, the dichloromethane and ethylacetate fractions showed the highest growth inhibitory effects on various cancer cells. For example, the dichloromethane and ethylacetate fractions ($200a{\mu}g/ml$) sig-nificantly inhibited the growth of various cancer cells by $89\∼96\%$ and$62\∼86\%$, respectively. DNA synthesis of AGS and Hep 3B cancer cells was significantly inhibited by adding dichloromethane fraction ($200{\mu}g/ml$) up to $94\%$ and $80\%$, respectively. Similarly, the ethylacetate fraction ($ 200\mug/ml $) showed a $ 95\% $ inhibition rate of DNA synthesis in AGS cells. These results suggest that the dichloromethane and ethylacetate fractions have specific active compounds, which will explain this anticancer effect of doenjang.

Characterization of Ovarian Cytochrome $P450_{C17}$ (17 ${\alpha}-hydroxylase$/17,20-lyase) in Rana dybowski (북방산 개구리 난소의 Cytochrome $P450_{C17}$ 유전자 특성)

  • Kang, Hae-Mook
    • Development and Reproduction
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    • v.10 no.2
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    • pp.127-133
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    • 2006
  • [ $17\;{\alpha}-hydroxylase/17,20-lyase(P450_{C17})$ ] is the key enzyme mediating the conversion of progesterone to $17\;{\alpha}-hydroxyprogesterone$, ultimately to androstenedione during steroidogenesis. R. dybowskii's ovarian $P450_{C17}$ cDNA was cloned to understand the regulatory mechanism of ovarian steroidogenic pathway at the molecular level in amphibian. A 2.5kb cDNA clone encoding a single open-reading frame with a 519 deduced amino acid was isolated with the screening of ovarian cDNA library. This sequence contained the three highly conserved domains as seen in $P450_{C17}$ of other species. The comparison of amino acid sequence of Rana $P450_{C17}$ with other animal's $P450_{C17}$ showed relatively high identity with 76% in Xenopus, 63% in chicken, 60% in rainbow trout, and 45% in human. Phylogenic analysis also indicated that Rana $P450_{C17}$ gene was evolutionary well conserved among vertebrate. Northern analysis indicated that the two different sizes of $P450_{C17}$ transcripts with approximately 2.5 and 3.6kb were detected in ovary tissue, but not in other tissues. The expression vector of Rana $P450_{C17}$ clearly showed the $17\;{\alpha}-hydroxylase$ activity converting the exogenous progesterone into $17\;{\alpha}-hydroxyprogesterone$ in the nonsteroidogenic COS-1 cells. Therefore, Rana $P450_{C17}$ cDNA is very useful to investigate the molecular mechanism of the ovarian steroidogenesis in amphibian.

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항암 및 항바이러스성 뉴크레오사이드의 합성

  • 이희주
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1994.04a
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    • pp.228-228
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    • 1994
  • Nucleoside계통의 유도체를 합성하여 체내 DNA 또는 RNA 합성대사에 영향을 줌으로써 궁극적으로 항암성 또는 항 virus성 작용을 나타내는 화합물을 얻고자 하였다. 본 연구에서는 천연의 nucleoside 구조중에서 특히 당부분에 변형을 준 일련의 acyclonucleoside 유도체들을 합성하였다. 이는 그간 많이 연구되어온acyclonucleoside 계열인 acyclovir계와는 달리 ribose 당부분을 $C_1$에서 $C_{5}$까지 거리가 유사한 acycloalkyl 결합형태로 결합하고 3'위치에 변형을 가져온 pyrimidine계 유도체를 합성하고자 일련의 반응을 시도하였다. 목적하는 화합물 계열을 얻지 못하고, 몇종류의 관련 유도체들을 분리하여 구조를 규명하고자 하였고, 이들중 일부에 대해 in vitro L1210 cell 증식억제효과를 검색하여 그 결과를 보고하고자 한다.

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