• Title/Summary/Keyword: DNA합성

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Effects of Denaturants on the Conditions of Polymerase Chain Reactions with G+C-rich Primers (G+C 함량이 높은 Primer를 사용하는 중합효소 연쇄반응에서 변성제가 미치는 영향)

  • 김종배;안준환;엄용빈;김영미
    • Biomedical Science Letters
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    • v.2 no.2
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    • pp.241-247
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    • 1996
  • Poor yields of amplified DNAs could be resulted in polymerase chain reaction(PCR) processes with G+C-rich DNA primers because of their high $T_m$ values. To maximize the yields of amplification in PCR processes with G+C-rich primers, we compared the yields of amplified DNA fragments according to the concentrations of specific denaturants added to the reaction mixture of PCR system. With addition of the mixture of 2.5% glycerol and 1.25% formamide, or 2.5% dimethyl sulfoxide to the reaction cocktail, respectively, remarkable increases in the yields of amplified DNA fragments were not observed in the PCR systems with G+C-low primers of Lyl chromosomal gene from Borrelia burgdorferi but observed in the PCR system with G+C- ich primers of Is900 gene from Mycobacterium parahberculosis. Although we were not practically able to discriminate the yields of PCR DNAs according to the concentrations used in this study, addition of the mixture of 5% glycerol and 2.5% formamide, or 5% DMSO tended to increase the production of extra bands.

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cDNA Cloning and Nucleotide Sequence Determination for VP7 Coding RNA Segment of Human Rotavirus Isolated in Korea (한국에서 분리된 사람 로타바이러스의 VP7 코딩 RNA 분절의 cDNA 합성과 염기서열 결정)

  • Kim, Young Bong;Kim, Kyung Hee;Yang Jai Myung
    • Korean Journal of Microbiology
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    • v.30 no.5
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    • pp.397-402
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    • 1992
  • The cDNA of RNA segment coding for VP7 of human rotavirus isolated from patient's stool at Seoul area was synthesized, amplified by polymerase chain reaction, field in with Klenow fragment of DNA polymerase I and cloned into pUC19. The cDNA sequence was determined and compared with that of VP7 coding RNA segments of group A rotaviruses isolates in foreign country. Over 90% sequence homology was found with serotyppe I sepcific WA1 and RE9 strains. Comparative analysis of the deduced amino acid sequences within the two variable regions (amino acid residue 87 through 101 and 208 through 221) with WA1 and RE9 strains also showed high degree of sequence similarity with each other.

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Effect of Extracts from Root Bark of Ulmus parvifolia on Inhibition of Growth and DNA Synthesis of Human Cancer Cells. (느릅나무 근피 추출물에 의한 인체 암세포 증식 및 DNA 합성 억제효과)

  • Lim, Sun-Young
    • Journal of Life Science
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    • v.17 no.9 s.89
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    • pp.1232-1236
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    • 2007
  • Growth and DNA synthesis inhibitory effects of extracts from root bark of Ulmus parvifolia on MG-63 human osteosarcoma cells, HT-29 human colon cancer cells and K-562 leukemia cancer cells were studied. The root bark extract of Ulmus parvifolia was extracted with methanol, hot water and juice. The methanol extract showed the highest inhibitory effect on growth of MG-63, HT-29 and K-562 cancer cells by >85%. The treatment of hot water and juice extracts from root bark of Ulmus parvifolia also inhibited growth of the above cancer cells with increasing concentration. DNA synthesis of MG-63 and HT-29 cancer cells was significantly inhibited by adding methanol, hot water and juice extracts from root bark of Ulmus parvifolia with increasing concentration, showing that the inhibitory effect of growth was more effective on HT-29 cancer cells. These results suggest that the methanol extract from root bark of Ulmus parvifolia may have specific active com-pounds on anticancer effect. The hot water extract also showed a strong inhibitory effect on growth of cancer cells, indicating that the active compounds may be stable to heat.

Analysis of Lambda Site-specific Recombination Inermediates Generated by Synthetic Substrates (합성 기질에 의해 형성된 Lambda Site-specific Recombination 중간 대사물의 분석)

  • 이나영;유승구
    • Microbiology and Biotechnology Letters
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    • v.23 no.3
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    • pp.282-287
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    • 1995
  • Integrase (Int) carries out the cutting and resealing of attachment (att) site DNA via a covalent Int-DNA intermediate. A family of synthetic substrate DNAs was designed to accumulate Int-DNA intermediate. Int-DNA intermediates accumulated by half substrate was analyzed by SDS- KCI precipitation and restriction digestion. The results showed that Int-half DNA intermediate was circular and contained covalently bound Int molecule. Int-DNA intermediates were also trapped with three other kinds of synthetic substrates.

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Automatic Acquisition of Local Fuzzy Rules by DNA Coding in new Composition Reasoning Method (새로운 합성 추론법에서 DNA 코딩을 이용한 국소 퍼지 규칙의 자동획득)

  • 박종규;안태천;윤양웅
    • Journal of the Korean Institute of Illuminating and Electrical Installation Engineers
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    • v.13 no.4
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    • pp.56-67
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    • 1999
  • In this paper, the new composition Irethod of global and local fuzzy reasoning concepts is proposed to reduce, optimize and automatically acquire the number of rules, without any lose of the general performances in conventional fuzzy controllers. In order to control the interaction between global reasoning and local reasoning, the DNA coding algorithm is introduced to the local fuzzy reasoning of the proposed composition fuzzy reasoning rrethod. The method is awlied to the real liquid level control system for the purpose of evaluating the performance. The sinru1ation results show that the proposed technique can control the system with higher accuracy and automatical1y acquire the fuzzy rules with rmre feasibility, than the conventional methods.ethods.

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Detection of DNA Sequence Polymorphism by Polymerase Chain Reaction in Fraxinus mandshurica Rupr Growing in Korea (P.C.R 기법(技法)을 이용(利用)한 들메나무 DNA sequence의 변이조사(變異調査))

  • Na, Chun Soo;Noh, Eun Woon;Kim, Young Joong;Shin, Chang Ho;Song, Won Sup;Kim, Sea Hyun
    • Journal of Korean Society of Forest Science
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    • v.81 no.4
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    • pp.320-324
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    • 1992
  • It has been reported that there are two distinct phenotypes in Fraxinus mandshurica Rupr. growing in Korea. Recently developed polymerase chain reaction(PCR) was used to detect DNA sequence polymorphism in the species. Using a thermostable DNA polymerase and synthetic DNA primers, unknown DNA sequences from the species were randomly amplified. The two types of the species produced different DNA amplification pattern with three different primers tested. Although DNA polymorphism was detected among individuals within types, each type has its own distinct pattern. The two types could be easily differentiated by trier characteristic predominant bands.

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Synthesis of 1-$\beta$-D-Arabinofuranosylcytosine-5'-methylthioacetate and Evaluation of Its Inhibitory Effect on DNA Synthesis (1-$\beta$-D-Arabinofuranosylcytosine-5'-methylthioacetate의 합성 및 이의 DNA 합성애 대한 억제작용 평가)

  • 이희주;송민경
    • YAKHAK HOEJI
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    • v.30 no.5
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    • pp.238-244
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    • 1986
  • As one of the starting materials, methylthioacetyl chloride(7) was synthesized in fair yield from mercaptoacetic acid via methyl methylthioacetate(5) prepared by alkylation employing N, N'-dicyclohexyl-O-methylisourea(4). Then 1-$\beta$-D-arabinofuranosylcytosine-5'-methylthioacetate (3) was prepared by esterification of ara-C with obtained methylthioacetyl chloride and tested for inhibitory activity on DNA synthesis in the growing primary hepatocytes and hepatoma strains($H_4$-II-E and HTC cells). In these in vitro cell lines, the inhibitory effect of ara-C-MTA(3) on DNA synthesis was similar to that of its parent ara-C but slightly lower.

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Oligonucleotide의 합성과 이용 <생명공학의 약학적 응용>

  • 나도선
    • YAKHAK HOEJI
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    • v.29 no.6
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    • pp.109-117
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    • 1985
  • 합성 DNA는 분자 생물학의 모든 분야의 연구에 사용될 수 있으며 앞으로도 그 이용도는 더욱 증가할 것으로 전망된다. 합성 기술의 발달로 생리활성을 가진 인조 유전자를 제조하는 것이 가능해졌으며 또한 다른 방법으로는 어려운 유전자의 분리와 cloning, site specific mutagenesis, 질병의 진단, 유전자의 구조 및 기능의 연구등 수많은 분야의 연구에 이용되고 있다. 본 고에서는 현재 여러 군데 분자 생물학 연구실에서 성공적으로 사용되고 있는 자동 합성기에 의한 phosphite합성법의 기초 이론과 합성된 oligonucleotide의 정제법에 대하여 간단히 서술하고 그 응용 방법에 대하여 논하고자 한다.

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