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        • KCI등재후보

          Cloning of Cytochrome P450 Gene involved in the Pathwayof Capsidiol Biosynthesis in Red Pepper Cells

          권순태,김재성,정도철,정정학,황재문,오세명 한국생명과학회 2003 생명과학회지 Vol.13 No.6

          In order to measure the enzyme activity of 5-epi-aristolochene hydroxylase, one of cytochrome P450 (P450) enzymes in eicitor-treated pepper cell, we used in vivo assay method and demonstrated a dramatic suppression of the activity by P450-inhibitors, ancymidol and ketocornazole. Using RT-PCR method with degenerate primer of the well conserved domains found within most P450-enzymes, and using cDNA library screening method, one distinct cDNA, being designated P450Hy01, was successfully isolated from elicitor-treated pepper cells. P450Hy01 mRNA was all induced in elicitor-treated cells whereas never induced in control cells. Moreover, levels of P450Hy01 expression were highly correlated with the levels of extracellular capsidiol production by different elicitors in cell cultures. P450Hy01 transcript was also induced by several other elicitors such as, cellulase, arachidonic acid, jasmonic acid, yeast extract as well as UV stress. P450Hy01 sequence contained high probability amino acid matches to known plant P450 genes and ORF with a conserved FxxGxRxCxG heme-binding domain. P450Hy01 cDNA showed 98% of homology in sequence of nucleotide as well as amino acid to 5-epi-aristolochene-1,3-hydroxylase (5EA1,3H) which has been isolated in tobacco cells, suggesting that P450Hy01 is prominent candidate gene for P450-enzyme encoding 5EA1,3H in pepper cell.Key words - red pepper, elicitor, heme-binding domain, cytochrome P450, 5-epi-aristolochene hydroxylase Capsidiol 생합성을 유도하는 Cytochrome P450 유전자의 탐색권순태*·김재성1·정도철2·정정학·황재문·오세명(*안동대학교 생명자원과학부, 1한국원자력연구소, 2상주대학교 자원식물학과)

        • KCI등재후보

          가지과식물에서 Capsidiol 생합성에 관여하는Cytochrome P450 유전자의 발현과 효소활성

          권순태,Paul Hasegawa 한국자원식물학회 2008 한국자원식물학회지 Vol.21 No.2

          '스콜라' 이용 시 소속기관이 구독 중이 아닌 경우, 오후 4시부터 익일 오전 7시까지 원문보기가 가능합니다.

          Enzyme activity and expression of cytochrome P450 gene involved in the pathway of capsidiol biosynthesis were compared in five different solanaceae plants such as red pepper, green pepper, tobacco, potato and egg plant. Base on genomic DNA and/or RT-PCR results, four solanaceae plants such as red pepper, green pepper, tobacco and egg plant possess P450 gene in the genome and specifically expressed by elicitor treatment. However, potato was appeared to have neither P450 nor cyclase gene in the genome. P450 genes did not show any expression in the plants under normal condition, but showed highly specific expression under elicitation condition in various organs and tissue such as leaf, root, stem and culture cells.

        • KCI등재

          시호의 약배양의 통한 배발생 및 재분화

          권순태,정형진,김길웅 한국생명과학회 2001 생명과학회지 Vol.11 No.4

          Effects of growth regulators(2,4-D, IAA, NAA, BA and kinetin) and chilling treatment on callus induction. embryogenesis and regeneration through anther cultures of B. falcatum L. were examined. Frequency of callus induction and embryogenesis was effectively increased by the treatment of chilling at 5$^{\circ}C$ for 10 days before anther inoculation. Optimal level of growth regulator for callus induction and embryogenesis from anther was 2,4-D 1.0 mg/L in Murashige and Skoog(MS) basal medium supplemented with 30 g/L sucrose, 8 g/L agar. Frequency of embryogenesis from anther derived callus was increased up to 48% or 45% by addition of IAA 0.1+ kinetin 1.0 mg/L of IAA 0.1+ BA 1.0 mg/L in MS medium, respectively, Optimal medium for obtaining green callus was MS basal supplemented with BA 1.0mg/L. Addition of auxins(IAA or NAA) inhibited the formation of green callus from anther derived callus.

        • KCI등재

          마늘유래 Cytochrome P450 유전자의 변이 분석

          권순태,Juli Kamiya 한국자원식물학회 2011 한국자원식물학회지 Vol.24 No.5

          '스콜라' 이용 시 소속기관이 구독 중이 아닌 경우, 오후 4시부터 익일 오전 7시까지 원문보기가 가능합니다.

          Wound inducible P450-Esg cDNA, one of cytochrome P450 gene family, was isolated from shoot of Euiseong garlic cultivar. P450-Esg cDNA possesses highly conserved heme-binding domain in the nucleotide sequence, and 1,419 bp of open reading frame (ORF) coding of 473 amino acids. Based on the nucleotide sequence analysis of P450-Esg homologous from twelve garlic cultivars, two domains, one domain between 472 to 510 bp, and the other between 1,210 to 1,249 bp from start codon (ATG), showed various nucleotide polymorphism among cultivars. Sequence of heme-binding domain in P450-Esg homologous, which is located at the domain between 1,210 to 1,240 bp from start codon, showed various nucleotide polymorphism as well as amino acid sequence polymorphism among twelve garlic cultivars. Anther domain, between 472 to 510 bp from start codon, showed exactly same amino acid sequence in the twelve garlic cultivars, but there were various single nucleotide polymorphism to the cultivars. 의성종 마늘의 유묘로부터 상처(wound)에 특이적으로 발현되는 cytochrome P450 유전자군의 하나인 P450-Esg cDNA를 탐색하였다. P450-Esg는 1,419 bp의 open reading frame(ORF)을 가지고 473개의 아미노산을 가진 polypeptide를 코딩하는 것으로 나타났다. 국내와 몽골로부터 수집한 12개의 재배종으로 부터 P450-Esg 유사 유전자의 염기서열을 비교한 결과 시작코돈(ATG)에서 472~510 bp 및 1,210~ 1,240 bp 부위의 염기에서 재배종간에 차이를 보이는 염기서열을 다수 확인하였다. cDNA 1,210~1,240 bp의 부위는 P450 유전자에서 공통적으로 알려진 heme binding domain으로, 각 지역에서 수집된 재배종은 염기서열뿐만 아니라 아미노산 서열에 있어서도 특이적인 변이를 보였다. cDNA 472~510 bp 부위에서 코딩하는 13개 아미노산의 서열은 12개 재배종에서 모두 동일하였으나, 13개 아미노산 중 7개에서 재배종 마다 각각 다른 DNA 염기로 코딩하는 단일염기다형성(single nucleotide polymorphism)을 보이는 서열을 확인하였다. 이 결과는 다양하게 존재하는 국내외 마늘 재배종을 구분하는 marker로 사용될 것이며, 외국산 마늘에 대한 유전적 우선권을 확보하는 수단으로 사용될 것이다.

        • KCI등재

          방사선처리가 고추의 생육과 항산화 효소의 활성에 미치는 영향

          권순태,정은아,김재성 한국생명과학회 2001 생명과학회지 Vol.11 No.6

          Seedlings of red pepper (Capsicum annum L.) were exposed to various doses of ${\gamma}$-radiation )($^{60}$ Co)and cultivated in the 1/2,000 a pot. Plant hight the number of leaves, flowers and fruits, chlorophyll contents, plant dry weight, ac- tivites of antiozidant enzymes such as superoxide dismutase(SOD) adn peroxidase(POD), were determined. Plant height of seedling treated with 1 Gy of ${\gamma}$-radiation was increased up to 16.6% as compared to untreated control at 80 days after treatment, while those treated with 20 Gy or 50 Gy was decreased 22.0% or 75.0% respectively. The plants treated with 1 Gy or 5 Gy were increased the number of leaves up to 41.2% and 24.9% as well as dry weight of fruit up to 58.4% and 49.4% respectively. However treatment of radiation higher than 20 Gy significantly inhibited the growth, chlorophyll content and yield of pepper fruit. Activities of antioxidant enzymes, SOD and POD were tem-porary increased within one or five days after treatment depending on the doses of treatment , and the activities were gradually decreased to control level of lower thereafter. The SOD and POD activities in the leaves of pepper plant treated with 50 Gy were 5.5 and 6.0folds higher than that of control at five days after treatment, respectively. 고추의 유모에 감마 방사선($^{60}$Co)을 조사하여 식물의 생육, 수량, 엽록소 함량및 항산화효소, superoxide dismutase (SOD) 및 peroxidase(POD)의 활성에 미치는 영향을 조사하였다. 고추의 초장은 1 Gy 처리에 의해 무처리보다 16.6%가 증가하였은 고선량인 20Gy 및 50Gy 처리에 의해서 는 각각 22.0 및 75.0%가 감소하였다. 잎의 수는 1Gy 및 5Gy처리에서 각각 41.2%및 24.9%가 증가하였으나 20Gy이상의 고선량에서는 감소하였고, 과실은 1및 5Gy 처리에의해 착과수가 증가하였고, 건물중 기준으로 각각 58.4및 49.4%가 증가하였다. 저선량의 방사선의 처리는 고추의 엽록소함량에 영향을 미치지 않았으나,증가된 활성은 다시 감소하여 처리후 20일에는 무처리와 비슥하거나 낮았다. 50Gy를 처리한 후 5일째에 측정한 SOD 및 POD의 효소 활성은 무처에 비하여 각각 5.5 및 6.0배가 증가하였다.

        • KCI등재

          애기장대 AtSIZ3 변이형의 온도 및 건조 스트레스에 대한 반응과 유전자 발현

          권순태,정형진,Paul Hasegawa 한국자원식물학회 2010 한국자원식물학회지 Vol.23 No.1

          '스콜라' 이용 시 소속기관이 구독 중이 아닌 경우, 오후 4시부터 익일 오전 7시까지 원문보기가 가능합니다.

          This study was carried out to understand the effect of low temperature(4℃), heat shock(37℃) and drought stresses on the growth and gene expression of Arabidopsis ATSIZ3(at1g08910) mutants. The seedling growth of SIZ3-mutants were markedly inhibited by the treatment of heat shock or chilling stresses. However, there was no significant differences between wild type and SIZ3-mutants in seeding fresh weight. As compared to wild type plants, SIZ3-mutants showed 63.9% inhibition of seedling fresh weight by the treatment of 10 days drought stress, suggesting that SIZ3 is involved in the resistance of Arabidopsis to drought stress. Base on RT-PCR analysis, expression of SIZ3 mRNA in the wild type showed 20% inhibition by chilling stress, 3.7 and 4.5 fold increase by the treatment of heat shock or drought stresses, respectively. 애기장대의 AtSIZ3(At1g08910) 유전자에 T-DNA를 삽입한 세 종류의 변이형에 저온(4℃), 고온(37℃) 및 건조스트레스를 처리하여 유묘의 생장반응과 유전자 발현을 조사하였다. 저온과 고온처리에 의해서는 야생형과 변이형간에 유묘생장에 유의한 차이를 보이지 않았다. 야생형과 변이형 식물체에 10일간의 건조스트레스를 처리하면 야생형은 재 관수에 의해 모든 식물체가 재생하였으나 변이형은 모두 고사하였고, 10일간의 건조처리로 변이형은 야생형에 비해 유묘생장이 평균 62.9%가 억제되는 것으로 나타났다. 야생형에서 AtSIZ3 유전자는 4℃의 저온처리에서 무처리를 보다 20%정도 발현이 감소하는 반면, 37℃ 고온처리에서는 3.7배, 건조처리에서는 4.5배가 증가하였다. 이 결과로 보아 AtSIZ3 유전자는 식물의 건조내성과 밀접한 연관이 있을 것으로 판단된다.

        • KCI등재

          고추(Capsicum annuum L.) 배양세포의 Elicitor 유도성 Phytoalexin 생성

          권순태,오세명 한국생명과학회 1999 생명과학회지 Vol.9 No.4

          Extracellular capsidiol, sesquiterpenoid phytoalexin, in the medium of pepper (Capsicum annuum L.) suspension cells was not identified from control cells, but highly accumulated in the elicitor-induced cells within 6 hours after the addition of 0.05$\mu\textrm{g}$/$m\ell$ cellulase. Capsidiol production in elicitor-induced cells was markedly suppressed by cytochrome P450 inhibitors, such as ancymidol and ketoconazole demonstrating that biosynthesis of capsidiol is catalyzed by at least on hydroxylation enzyme in the biochemical pathway. Based on protein electrophoresis, two bands, 23.0kDa and 27.5kDa, were identified as newly synthesized polypeptides in the elicitor-induced suspension cells, suggesting that pepper cells which were subjected to elicitor treatment activate specific gene(s) for capsidiol biosynthesis in cultured cells.

        • KCI등재

          Superoxide dismutase의 활성차이에 따른 식물세포의 paraquat에 대한 반응과 핵 DNA 손상 검정

          권순태,이명현,오세명,정도철,김길웅 한국생명과학회 2004 생명과학회지 Vol.14 No.4

          This study was undertaken to investigate the different responses of cultured plant cells to paraquat treatment and nucleus-DNA damage in relation to enzyme activity of superoxide dismutase (SOD). Furthermore, this study was also carried out to understand the antioxidative mechanism of plant cells to environmental stress. We selected two different species of plant cultured cells, Ipomoea batatas as high-SOD species and Lonicera japonica as low-SOD species. The total activity and specific activity of SOD in a chlorophyllous cell of I. batatas were 3,736 unit/gㆍfresh weight and 547 unit/mgㆍprotein, respectively, and those in L. japonica were 23 unit/gㆍfresh weight and 13 unit/mgㆍprotein, respectively SOD activity in chlorophyllous I. batatas cells reached its maximum level at 10 to 15 days after subculture, whereas that in L. japonica remained at a very low SOD level during the whole period of subculture. In comparison to L. japonica, I. batatas, a high-SOD species, showed high tolerance to paraquat 10 and 50 mg/l treatment in terms of cell viability and electrolyte leakage. Based on the result of comet assay, the nucleus-DNA damage of two species by paraquat 50 mg/l treatment was not significantly different. However, I. batatas cells repaired their damaged DNA more effectively than the cells of the low-SOD species, L. japonica.

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