• Title/Summary/Keyword: DNA 염기 서열

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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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Cloning of SNAS-25 Gene from Rat Brain cDNA Library (Rat Brain cDNA Library로부터 SNAP-25 유전자의 클로닝)

  • Cho, Ae-Ri;Ji, Young-Mi;Yoo, Min;Lee, Soon-Chul;Yoo, Kwan-Hee
    • Biomedical Science Letters
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    • v.6 no.1
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    • pp.11-17
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    • 2000
  • SNAP-25 was first investigated as a neuron-specific protein preferentially expressed in CA3 pyramidal neurons of mouse hippocampus. It is a presynaptic plasma membrane protein in the nerve cell and plays an important role in the synaptic vesicle membrane docking and fusion pathway. We have recently isolated SNAP-25 cDNA from a rat brain cDNA library using a probe of Z2 cDNA. It consisted of 2,101 bp and an open reading frame (ORF) was identified between nucleotides (nt) 209 and 827. The AUG codon (nt 209∼211) was surrounded by CTACCATGG, which corresponded to the consensus sequence of ribosomal binding site. The ORF was terminated by TAA (nt 827∼829) to encode a polypeptide of 206 amino acid residues. The 3'-untranslated region contained two extensive stretches of repeated (CA)28 and (CA)19 at positions 925∼980 and 1645∼1682. It is noteworthy that cysteine residues were clustered in the span of amino acid residues 84∼991 : Cys-Gly-Leu-Cys-Val-Cys-Pro-Cys. Rat SNAP-25 showed 88% and 97% identity in nucleotide sequences to that of human and mouse, respectively. Amino acid sequence of rat SNAP-25 showed 100% identity to that of mouse and human SNAP-21.

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Identification of Arbuscular Mycorrhizal Fungi Colonizing Panax ginseng Using 18S rDNA Sequence (18S rDNA를 이용한 인삼(Panax ginseng)의 내생균근 균의 동정)

  • Eo, Ju-Kyeong;Kim, Dong-Hun;Jeong, Hyeon-Suk;Eom, Ahn-Heum
    • Applied Biological Chemistry
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    • v.47 no.2
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    • pp.182-186
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    • 2004
  • Morphological observation of roots and molecular technique were used to investigate the symbiotic relationships between arbuscular mycorrhizal (AM) fungi and ginseng roots. Korean ginseng, Panax ginseng, was collected from 8 sites in Korea. Colonization pattern of AM fungi in ginseng roots was determined as an Arum type under light microscopes. Nested PCR using AM fungal specific primers was employed to amplify a partial region on 18s rDNA of AM fungi from the root extracted mixed DNA. The amplified DNA was cloned and analyzed by random fragment length polymorphism (RFLP) with restriction enzymes, AluI, HinfI and AsuC21. One from each RFLP pattern was selected for sequencing. A total 16 clones were sequenced and identified as 2 species of AM fungi; Paraglomus brasilianum and Glomus spurcum. Paramglomus brasilianum was found from most of the ginseng roots, in this syudy suggesting that this species of AM fungi could have specific relationship with the ginseng root. Possible roles of AM fungal species in ginseng roots are discussed.

Genetic relationship of Aloe vera 'Saengjang', a new forma, based on cpDNA and ITS sequence variation (cpDNA와 ITS 염기변이에 근거한 신품종 생장알로에 유전적 상관관계)

  • Srikanth, Krishnamoorthy;Jang, Seon Il;Whang, Sung Soo
    • Korean Journal of Plant Taxonomy
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    • v.44 no.4
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    • pp.250-256
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    • 2014
  • This study was carried out to understand the genetic relationship of three Aloe spp. cultivated in Korea, A. saponaria, A. vera and A. arborescens and a new variant in Korea based on three plastid (matK, trnL-F, rbcL) and one nuclear (ITS regions) DNA barcode markers. A total of 2,420 bp sequence was amplified. Two indels were detected in the trnL region, and also several species specific nucleotide loci were detected in all 29 parsimonious informative sites, and 148 variable sites were detected among four taxa studied while 170 variable and 75 parsimonious sites were detected when other Aloe spp. in worldwide were used. An UPGMA phenogram with 10,000 bootstrap replication showed that the new variant was closest to A. vera. The variant was not morphologically and genetically concurrent with any reported species so far. The clustering of Aloe species were broadly in agreement with previously reported results.

Analysis of ITS DNA Sequences of the Viola albida Complex (태백제비꽃군 ITS DNA 염기서열 분석)

  • Whang, Sung-Soo
    • Korean Journal of Plant Resources
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    • v.19 no.5
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    • pp.628-633
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    • 2006
  • ITS DNA sequences from five individuals, representative of five groups designated according to the degree of leaf teeth and lobes from simple to palmate compound leaf in the Viola albida complex, established and further analysed in order to solve the taxonomic difficulty. A total 702 bp was sequenced at the 5.8S ribosomal DNA and internal transcribed spacer 1 and 2. The 5.8S coding region is 163 bp, and has no sequence variations. The ITS1 and ITS2 noncoding regions have a little bit sequence variations, and those were further analysed by the methods of the analysis of variance (ANOVA), the analysis of sequence divergence and the phylogenetic analysis. The result of ANOVA showed no significant differences among individuals investigated. The analysis of sequence divergence with Kimura 2-parameter distance revealed that in-groups showed much less than 0.05 in absolute value among individuals, while two out groups more than 0.05, V. grypoceras and V. orientalis. This result appeared that the sequence divergence among in-groups was not yet occurred in the species level but situated at somewhere below the species level. In the phylogenetic analysis, two outgroups formed the basal clades in order. Five individuals in-groups formed a clade. The clade was, however, not very robust as around 50% in bootstrap value, suggesting that this result was not meaningful in the phylogenetic point of views.

Identification of Bacteria Causing Fermentation of Oriental Melon in Korea (참외 발효과를 유발하는 세균의 동정)

  • Choi, Jae-Eul;Cha, Sun-Kyung;Kim, Jin-Hee;Yuk, Jin-Ah;Hwang, Yong-Soo;Kwon, Soon-Wo
    • Research in Plant Disease
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    • v.9 no.4
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    • pp.189-195
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    • 2003
  • Bacteria causing fermentation in Oriental melon were identified as three independent groups on the basis of 16S rDNA sequence analysis. The 16S rDNA sequence of the strain CM2105 showed the highest identity (99.6%) with that of Microbacterium phyllosphaerae, and also indicated high sequence identity to that of M. holiorum (99.5%). The 16S rDNA sequences of the strain CM2101 and CM2121 matched at the high sequence similarity (98.9%, 98.8, respectively), to that of Pseudomonas pavonacea, and the DNA sequence of CM2126 showed high sequence identity to that of P. costantinii (99.5%), and P. grimontii (99.0%). The 16S rDNA sequence of the strain CM2113 showed the highest identity (99.7%) with that of Enterobacter cloacae. The 16S rDNA sequences, the physiological and biochemical analysis suggested that the strain CM2105 belonged to Microbacterium phyllosphaerae, CM2101, CM2121 and CM2126 to Pseudomonas spp., CM2113 to Enterobacter cloacae.

Korea Brassica Genome Project: Current Status and Prospective (배추 유전체열구의 현황과 전망)

  • Choi, Su-Ryun;Park, Jee-Yong;Park, Beom-Seok;Kim, Ho-Il;Lim, Yong-Pyo
    • Journal of Plant Biotechnology
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    • v.33 no.3
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    • pp.153-160
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    • 2006
  • Brassica rape is an important species used as a vegetable, oil, and fodder worldwide. It is related phylogenically to Arabidopsis thaliana, which has already been fully sequenced as a model plant. The 'Multinational Brassica Genome Project (MBGP)'was launched by the international Brassica community with the aim of sequencing the whole genome of B. rapa in 2003 on account of its value and the fact that it has the smallest genome among the diploid Brassica. The genome study was carried out not only to know the structure of genome but also to understand the function and the evolution of the genes comprehensively. There are two mapping populations, over 1,000 molecular markers and a genetic map, 2 BAC libraries, physical map, a 22 cDHA libraries as suitable genomic materials for examining the genome of B. rapa ssp. pekinensis Chinese cabbage. As the first step for whole genome analysis, 220,000 BAC-end sequences of the KBrH and KBrB BAC library are achieved by cooperation of six countries. The results of BAC-end sequence analysis will provide a clue in understanding the structure of the genome of Brassica rapa by analyzing the gene sequence, annotation and abundant repetitive DHA. The second stage involves sequencing of the genetically mapped seed BACs and identifying the overlapping BACs for complete genome sequencing. Currently, the second stage is comprises of process genetic anchoring using communal populations and maps to identify more than 1,000 seed BACs based on a BAC-to-BAC strategy. For the initial sequencing, 629 seed BACs corresponding to the minimum tiling path onto Arabidopsis genome were selected and fully sequenced. These BACs are now anchoring to the genetic map using the development of SSR markers. This information will be useful for identifying near BAC clones with the seed BAC on a genome map. From the BAC sequences, it is revealed that the Brassica rapa genome has extensive triplication of the DNA segment coupled with variable gene losses and rearrangements within the segments. This article introduces the current status and prospective of Korea Brassica Genome Project and the bioinformatics tools possessed in each national team. In the near future, data of the genome will contribute to improving Brassicas for their economic use as well as in understanding the evolutional process.

Molecular Cloning and Alternative Splicing of Growth Hormone Transcripts in Greenling, Hexagrammos otakii (쥐노래미 (Hexagrammos otakii) 성장호르몬 cDNA유전자의 염기서열 변이 및 발현 특성)

  • Nam Yoon Kwon;Kim Dong Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.35 no.6
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    • pp.676-681
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    • 2002
  • Different types of transcripts encoding growth hormone (GH) were identified from cDNA libraries constructed with pituitaries of a marine fish species, greenling (Hexagrammos otakii). GH-homologous cDNA clones were isolated using the high-density filter hybridization and the expressed sequence tag techniques. Of 39 full-length positive cDNA clones, 31 clones ($79\%$) displayed an identical sequence, however, remaining 8 clones exhibited several polymorphisms in their sequences including (1) the length and sequence variability in the 5' upstream region, (2) insertional sequences in open reading frame, and (3) deletion and/or single nucleotide polymorphism in the untranslated 3' region. Based on RT-PCT and RNA dot blot analyses, these transcripts were proven to be expressed in a pituitary-specific manner.

Code Optimization of DNA Computing for Travelling Salesman Problem (Travelling Salesman Problem을 위한 DNA 컴퓨팅의 코드 최적화)

  • Kim, Eun-Kyoung;Lee, Sang-Yong
    • Annual Conference of KIPS
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    • 2002.11a
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    • pp.323-326
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    • 2002
  • DNA 컴퓨팅은 생체 분자들이 갖는 막대한 병렬성을 이용하여 조합 최적화 문제에 적용하는 연구가 많이 시도되고 있다. 특히 TSP(Travelling Salesman Problem)는 간선에 대한 가중치 정보가 추가되어 있기 때문에 가중치를 DNA 염기 배열로 표현하기 위한 효율저인 방법들이 제시되지 않았다. 따라서 본 논문에서는 DNA 컴퓨팅에 DNA 코딩 방법을 적용하여 정점과 간선을 효율적으로 생성하고 표현된 DNA 염기 배열의 간선에 실제간을 적용하여 가중치 정보를 계산하는 ACO(Algorithm for Code Optimization)를 제안한다. DNA 코딩 방법은 변형된 유전자 알고리즘으로 DNA 기능을 유지하며, 서열의 길이를 줄일 수 있으므로 최적의 서열을 생성할 수 있는 특징을 갖는다. 실험에서 ACO를 TSP에 적용하여 Adleman의 DNA 컴퓨팅 알고리즘과 비교하였다. 그 결과 초기 문제 표현에서 우수한 적합도 값을 생성했으며, 경로의 변화에도 능동적으로 대처하여 최적의 결과를 빠르게 탐색할 수 있었다.

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Phylogenetic Analysis of Ji-Mo (Anemarrhena asphodeloides) on the Basis of Chloroplast DNA Sequences (엽록체 DNA 염기서열을 이용한 한약재 지모의 기원 확인 및 유연관계 분석)

  • Kim, Myung-Kyum;Jigden, Baigalmaa;Sun, Hua;Noh, Jong-Hun;Kim, Se-Young;Yang, Deok-Chun
    • Korean Journal of Medicinal Crop Science
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    • v.16 no.1
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    • pp.20-26
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    • 2008
  • Anemarrhena asphodeloides (Korean name "Ji-Mo") has been used for oriental medicinal purposes in Korea, China and Japan. In this study, 29 A. asphodeloides samples were collected including 3 certified A. asphodeloides plants and many commercially marketed A. asphodeloides products. Chloroplast trnL-F regions of the "Ji-Mo" samples were sequenced and used to identify whether the samples were genuine A. asphodeloides or not. As the result, the trnL-F sequences of all the "Ji-Mo" samples were shown to be identical and it was proven that commercially available medicinal products "Ji-Mo" are genuine A. asphodeloides. Phylogenetic tree of. A. asphodeloides using the trnL-F sequences was constructed and compared with phylogenetic tree using rubisco large subunit (rbcL) gene sequences. In these tree, A. asphodeloides was affiliated in the family Agavaceae in the order Asparagales. It is proven that trnL-F phylogenetic tree is useful to study taxonomic position of A. asphodeloides.