• 제목/요약/키워드: DNA separation

검색결과 121건 처리시간 0.026초

A Molecular Phylogenetic Study on Korean Alexandrium catenella and A. tamarense Isolates (Dinophyceae) Based on the Partial LSD rDNA Sequence Data

  • Kim, Keun-Yong;Kim, Chang-Hoon
    • Journal of the korean society of oceanography
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    • 제39권3호
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    • pp.163-171
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    • 2004
  • Sequences of the large subunit ribosomal (LSD) rDNA D1-D2 region of Alexandrium catenella(=A. sp. cf. catenella) and A. tamarense isolates, which were collected along the Korea coasts, were analyzed to understand their phylogenetic relationships and geographical distributions. All A. catenella and A. tamarense isolates belonged to the A. tamarense/catenella/fundyense complex and were grouped with the North American and temperate Asian ribotypes, respectively, regardless of the presence or absence of a ventral pore in the first apical plate. A consistent and peculiar characteristic that differentiated the Alexandrium isolates was amplification of a second PCR product with a lower molecular weight in addition to the predicted one; ten A. catenella isolates belonging to the temperate Asian ribotype yielded this additional PCR product. Sequence alignment revealed that the shorter PCR product resulted from an unusual large deletion of 87 bp in the LSD rDNA D1 domain. The North American and temperate Asian ribotypes were prevalent along the Korean coasts without geographical separation. Given the high genetic homogeneity among widely distributed Alexandrium populations, each ribotype appeared to be pandemic rather than to constitute a distinct regional population.

안정성 및 Transfection 효율이 우수한 양이온성 리포좀 유전자 전달시스템의 개발 (New Cationic Liposome with Enhanced Stability and Transfection Efficiency for Gene Delivery)

  • 김경미;남방현;손동환
    • Journal of Pharmaceutical Investigation
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    • 제28권2호
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    • pp.93-98
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    • 1998
  • We have developed liposomes which can be easily prepared with inexpensive lipid, have enhanced stability, and can efficiently deliver DNA into the COS-l cells, Liposome formulations were prepared using cationic materials such as dimethyldioctadecyl ammonium bromide (DDAB), cetyltrimethyl ammonium bromide(CTAB), We investigated the effect of cationic liposome formulations on in vitro DNA transfection, DDAB-containing liposomes showed increased transfection efficiency which was 3.2-fold as much as that by $Lipofectin^{\circledR}$, but CTAB-containing liposomes were inactive in gene transfection. The effect of colipid of DDAB-containing liposome was also investegated. As a colipid, dioleylphosphatidylethanolamine(DOPE) and cholesterol did altered the transfection efficiency of DDAB-containing liposomes. And increased DDAB concentration lowered the transfection efficiency. The optimum amount of liposomal formulation was $10\;{\mu}M$ for $1\;{\mu}g$ of DNA. In the experiment of stability, DOPE-containing liposomes formulation showed a broad size distribution and separation of two major peaks on a 5th day of preparation, but liposomes containing cholesterol was stable for 10 days. DDAB-containing liposomal DNA delivery system was prepared easily and was stable.

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Streptomyces levendulae의 원형질체와 원형질체 융합에 대한 미세구조 (Ultrastructural Studies for Protoplasts and Protoplast Fusion in Streptomyces lavendulae)

  • 하영칠;홍순우;유진철;임헌만
    • 미생물학회지
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    • 제24권3호
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    • pp.197-203
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    • 1986
  • Streptomyces lavendulae에서 생성된 원형질체 및 원형질체 융합과정에 대한 미세구조 및 행태학적 관찰을 투과 전자현미경과 주사전자현미경을 사용하여 행하였다. 생성된 원행질체들은 고장액에서 대체로 안정하였으나 간혹 변형된 원형질체들이 관찰되었다. 원형질체 융합은 접촉지역 융합지역, 분리지역 형성과정을 차례로 거치면서 선행되는 것이 관찰되었다. 이러한 사실은 세포막 구조의 변화와 이원형의 소실에 의하여 뒷받침이 될 수 있으으로 이러한 변화들을 융합과정에서의 각 단계로 해석하는 것이 가능하리라 사료된다.

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12-Oxoeicosatetraenoic acid, a candidate signal for placenta separation, activates matrix metalloproteinase and induces apoptosis in bovine trophoblast cells

  • Hachiro Kamada
    • Animal Bioscience
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    • 제36권3호
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    • pp.429-440
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    • 2023
  • Objective: 12-oxo-5Z,8Z,10E,14Z-eicosatetraenoic acid (12-KETE), a metabolite of arachidonic acid, is a strong candidate signal for placenta separation following calf discharge at delivery. In the present study, the effects of 12-KETE on bovine trophoblast cells were investigated to determine its function in the placentome at delivery. Methods: Bovine trophoblast cells derived from blastocysts were used. They were cocultured with or without fibroblasts derived from bovine placentome and/or bovine uterine epithelial cells. 12-KETE was added to the culture medium. Results: Bovine trophoblast cells contained binucleate cells and strongly expressed caudal type homeobox 2 (CDX-2) genes. Addition of 12-KETE to the trophoblast cell colony without feeder cells or that on a fibroblast monolayer induced rapid exfoliation of the colony. After 12-KETE addition, trophoblast cells emitted strong fluorescence caused by the degradation of dye-quenched collagen, indicating that 12-KETE activated matrix metalloproteinase of the trophoblast cells. Exfoliated cell colonies were stained with YOPRO-1, but not propidium iodide (PI). Moreover, DNA fragmentation and Bcl-2 associated X protein (Bax) gene (apoptosis stimulator) upregulation were observed in exfoliated cells, indicating that 12- KETE induced trophoblast cell apoptosis. These results were consistent with previous in vivo observations; however, even a lower concentration of 12-KETE activated trophoblast protease. Meanwhile, fibroblasts derived from the bovine placentome converted arachidonic acid to 12-KETE. Conclusion: These observations indicate that 12-KETE may serve as a signal for placenta separation at delivery.

Region Identification on a Trained Growing Self-Organizing Map for Sequence Separation between Different Phylogenetic Genomes

  • Reinhard, Johannes;Chan, Chon-Kit Kenneth;Halgamuge, Saman K.;Tang, Sen-Lin;Kruse, Rudolf
    • 한국생물정보학회:학술대회논문집
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    • 한국생물정보시스템생물학회 2005년도 BIOINFO 2005
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    • pp.124-129
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    • 2005
  • The Growing Self-Organizing Map (GSOM), an extended type of the Self-Organizing Map, is a widely accepted tool for clustering high dimensional data. It is also suitable for the clustering of short DNA sequences of phylogenetic genomes by their oligonucleotide frequency. The GSOM presents the result of the clustering process visually on a coloured map, where the clusters can be identified by the user. This paper describes a proposal for automatic cluster detection on this map without any participation by the user. It has been applied with good success on 20 different data sets for the purpose of species separation.

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애너그램 문제 인지적 해결과정의 분자컴퓨팅 시뮬레이션 (Molecular Computing Simulation of Cognitive Anagram Solving)

  • 천효선;이지훈;류제환;백다솜;장병탁
    • 정보과학회 컴퓨팅의 실제 논문지
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    • 제20권12호
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    • pp.700-705
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    • 2014
  • 애너그램은 주어진 문자들을 재배열하여 숨겨진 단어를 찾아내는 철자바꾸기 놀이로, 문제를 빨리 풀어내는 사람들은 제약 만족 네트워크의 병렬적 탐색에 의해 문제를 해결한다. 본 연구에서는 이러한 인지적 현상을 모델링한 분자 애너그램 풀이 알고리즘을 제시하였다. 문자를 DNA 서열로 인코딩하고, 문자 DNA 가닥을 연결하여 바이그램과 단어 서열을 만들었다. DNA 혼성화, 연결, 젤 전기영동, 추출 연산을 수행해 문자와 바이그램 집합으로부터 답을 찾는 데 필요한 바이그램을 추출한 후, 추출한 바이그램과 단어 집합으로부터 다시 네 가지 DNA 연산을 반복하여 답을 찾는다. 분자 실험 결과 분자 컴퓨터는 정답인 단어와 오답인 단어를 구분해낼 수 있었다. 이를 통해 인간의 병렬적 사고과정을 분자 컴퓨터로 모델링할 수 있는 가능성을 보였다.

한국고유종 북방종개(어류강, 미꾸리과)의 집단유전학적 구조 (Population Genetic Structure of the Korean Endemic Species, Iksookimia pacifica (Pisces: Cobitidae) Distributed in Northeast Korea)

  • 장숙진;고명훈;권예슬;원용진
    • 한국환경생태학회지
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    • 제31권5호
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    • pp.461-471
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    • 2017
  • 북방종개(Iksookimia pacifica)의 유전적 다양성과 구조적 특징을 밝히기 위해 동해 독립하천들에 서식하는 10개의 집단들을 대상으로 핵유전자와 미토콘드리아 유전자에 기반한 집단유전학적 분석을 실시하였다. 일부 예외적인 경우를 제외하고, 미토콘드리아와 핵유전자 모두 통계적으로 유의미한 집단 간 유전적 분화가 관찰되었다. 핵유전자들의 DNA 서열자료에서 추출한 유전자형 자료를 Bayesian 방법으로 분석한 결과 북방종개는 천진천과 양양남대천을 기준으로 북쪽과 남쪽의 두 개의 그룹으로 나뉘는 구조를 보였다. 현재 동해 하천들이 지리적으로 단절되어 온 독립 수계라는 것을 감안했을 때, 남북으로 구별되는 집단유전적 구조는 북방종개가 한반도에 정착했던 초기 조상 집단이 남북으로 갈라지는 지리적인 분리 사건과 관련되었을 것으로 해석되며, 이러한 초기 조상집단의 지리적 분리 이후, 두 조상그룹들은 남북의 지리적인 범위 내에서 하천 별 고립에 따른 추가적인 분화 과정을 거쳤을 것으로 추정된다. 주목할 점으론, 자산천 집단의 많은 개체들이 지리적 거리가 먼 양양남대천 및 강릉남대천 집단과 하나의 유전적 cluster를 형성하고 있는 것이다. 이와 함께 미토콘드리아 유전자의 경우 몇몇 이웃하는 집단들 사이에 현저히 낮은 유전적 분화도 그리고 일부 집단들에서 매우 낮은 유전적 다양성이 관찰되었다. 본 집단유전학적 결과는 향후 북방종개의 보존 및 관리를 위한 기초자료로 제시될 것이다.

Comparison of Endonuclease-Sensitive Sites by T4 Endonuclease V and UvrABC Nuclease Treatments Followed by Formamide or Sodium Hydroxide Denaturation

  • Chang, Yung-Jin
    • BMB Reports
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    • 제31권4호
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    • pp.405-408
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    • 1998
  • Endonuclease-sensitive sites detected by T4 endonuclease V or UvrABC nuclease treatments were compared in the dihydrofolate reductase gene of UV-irradiated Chinese hamster ovary B-11 cells. The number of endonuclease-sensitive sites detected by T4 endonuclease V treatment followed by NaOH denaturation was twice that of formamide denaturation. Repeated treatment of damaged genomic DNA with T4 endonuclease V resulted in no further increase in the number of endonuclease-sensitive sites detected. The numbers of endonuclease-sensitive sites detected by UvrABC nuclease using each denaturation condition were similar. Sequential treatment with the two endonucleases using formamide denaturation resulted in twice the number of endonuclease-sensitive sites detected by treatment of each nuclease alone. Due to a lack of AP endonuclease activity these results suggest the presence of T4 endonuclease V-sensitive sites which could be complemented by alkaline gel separation or by UvrABC nuclease treatment.

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토양으로부터 genomic DNA의 효과적인 분리 (Improved Genomic DNA Isolation from Soil)

  • 강주형;김보혜;이선이;김영진;이준원;박영민;안순철
    • 생명과학회지
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    • 제15권6호
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    • pp.851-856
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    • 2005
  • Although valuable microbes have been isolated from the soil for the various productions of useful components, the microbes which can be cultivated in the laboratory are only $0.1-1\%$ of all microbes. To solve this problem, the study has recently been tried for making the valuable components from the environment by directly separating unculturable micrbial DNA in the soil. But it is known that humic acid originated from the soil interrupts various restriction enzymes and molecular biological process. Thus, in order to prevent these problems, this study modified the method separated soil DNA with phenol, CTAB and PEG. In order to compare the degree of purity for each DNA and the molecular biological application process, $A_{260}/A_{280}$ ratio, restriction enzymes, and PCR were performed. In case of DNA by the modified method, total yield of DNA was lower but $A_{260}/A_{280}$ ratio was higher than the previously reported methods. It was confirmed that the degree of purity is improved by the modified method. But it was not cut off by all kinds of tested restriction enzymes because of the operation of a very small amount of interrupting substances. When PCR was operated with each diluted DNA in different concentrations and GAPDH primer, the DNA by the modified method could be processed for PCR in the concentration of 100 times higher than by the previously reported separation method. Therefore, this experiment can find out the possibility of utilization for the unknown substances by effectively removing the harmful materials including humic acid and help establishing metagenomic DNA library from the soil DNA having the high degree of purity.

Molecular cloning, tissue distribution and quantitative analysis of two proopiomelanocortin mRNAs in Japanese flounder (Paralichthys olivaceus)

  • Kim, Kyoung-Sun;Kim, Hyun-Woo;Chen, Thomas T.;Kim, Young-Tae
    • BMB Reports
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    • 제42권4호
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    • pp.206-211
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    • 2009
  • Proopiomelanocortin (POMC) plays an essential role in the stress response of the hypothalamic-pituitary-adrenal axis, and is the precursor of biologically active peptides such as adrenocorticotropin (ACTH), $\alpha$-melanocyte-stimulating hormone ($\alpha$-MSH), $\beta$-melanocyte-stimulation hormone ($\beta$-MSH) and $\beta$-endorphin. We have synthesized two different forms of POMC cDNA clones, POMC-I and POMC-II, from a pituitary cDNA library for Paralichthys olivaceus, or Japanese flounder. jfPOMC-I cDNA consists of 954bp and encodes a polypeptide of 216 amino acid residues, whereas jfPOMC-II consists of 971bp which encode a polypeptide of 194 amino acid residues. The high levels of jfPOMC-I and -II mRNAs detected in the pituitary tissue and moderate levels detected in the brain tissue plus our quantitative RT-PCR analysis, which showed there to be no significant difference between the levels of jfPOMC-I and -II mRNAs, indicate that there may be no functional separation between these two mRNAs in the flounder.