• 제목/요약/키워드: DNA-dependent

검색결과 1,346건 처리시간 0.032초

Determination of the minimal sequence of bovine lactoferricin responsible for apoptosis induction

  • Yoo, Yung-Choon;Lee, Kyung-Bok;Lee, Hoi-Young
    • 대한약학회:학술대회논문집
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    • 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-2
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    • pp.134.2-135
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    • 2003
  • We examined the minimal amino acid sequence of bovine lactoferricin (Lfcin-B), a cationic peptide corresponding to residues 17-41 near the N-terminus of bovine lactoferrin, to induce apoptosis in THP-l human monocytic leukemic cells using synthetic peptides. A synthetic peptide (Lfc-17/29, amino acid sequence; FKCRRWQWRMKKL) which is consist of 13 amino acids near the N-terminus of Lfcin-B induced cell death in THP-1 cells in a dose-dependent manner, showing apparent apoptotic changes such as hypodiploid forms of genomic DNA and apoptotic DNA fragmentation. (omitted)

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환경호르몬에 의해 손상된 골모세포에 대한 인삼의 영향 (Effects of Ginseng Radix Against Osteoblasts Damaged by Endocrine Disrupting Chemicals)

  • 홍기연;박승택;전병훈;서은아
    • 동의생리병리학회지
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    • 제16권2호
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    • pp.338-342
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    • 2002
  • In order to evaluate the effect of Ginseng Radix(GR) against endocrine disrupting chemicals(EDC), cultured mouse osteoblasts were preincubated with various concentrations of GR extract before the exposure of Bisphenol A for 12 hours. Cytotoxic effect of Bisphenol A was measured by the XTT assay. In addition, the protective effect of GR over Bisphenol A-induced cytotoxicity on osteoblasts was assessed by the DNA and protein synthesis in these cultures. The results were as follows : Osteoblastic cell viability was decreased in dose and time dependent manner after exposed to various concentrations of Bisphenol A. Midcytotoxicity value(MCV50) of Bisphenol A was determined at 6μM Bisphenol A after osteoblasts were grown for 12 hours in the media containing various concentrations of Bisphenol A. Amount of DNA synthesis was increased in dose-dependent manner after cultured osteblasts were pretreated with GR for 2hrs before exposure to Bisphenol A for 12 hours. Amount of protein synthesis was increased in dose-dependent manner after cultured osteoblasts were pretreated with GR for 2 hours before exposure of Bisphenol A for 12 hours. From these results, it is suggested that Bisphenol A was highly toxic by the decrease of the cell viability, and GR is effective in the prevention of Bisphenol A-induced cytotoxicity by the increase of DNA and protein syntheses in cultured mouse osteoblasts.

RNAseq 빅데이터에서 유전자 선택을 위한 밀집도-의존 정규화 기반의 서포트-벡터 머신 병합법 (Combining Support Vector Machine Recursive Feature Elimination and Intensity-dependent Normalization for Gene Selection in RNAseq)

  • 김차영
    • 인터넷정보학회논문지
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    • 제18권5호
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    • pp.47-53
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    • 2017
  • 고처리 시퀀싱과 빅데이터 및 크라우드 컴퓨팅에 혁신이 일어나면서, RNA 시퀀싱도 획기적인 변화가 일어, RNAseq가 기존의 DNA 마이크로어레이를 대체하여, 빅-데이터를 형성하고 있다. 현재, RANseq 이용한 유전자 조절망(GRN) 까지 연구가 활성화 되고 있는데, 그 중 한 분야가 GRN의 기본 요소인 특징 유전자를 빅-데이터에서도 구별하고 기존에 알려진 것 외에 새로운 역할을 찾는 것이다. 그러나, 이러한 연구 방향에 부합하는 빅-데이터를 처리할 수 있는 컴퓨테이션 방법이 아직까지 매우 부족하다. 따라서 본 논문에서는 RNAseq 빅-데이터를 처리할 수 있도록 기존의 SVM-RFE알고리즘을 밀집도-의존 정규화에 병합하여, NCBI-GEO와 같은 빅-데이터에서 공개된 일부의 데이터에 개선된 알고리즘을 적용하고 해당 알고리즘에 의해 나온 결과의 성능을 평가한다.

Apoptotic Activity of Insect Pathogenic Fungus Paecilomycesc japonica Toward Human Acute Leukemia Jurkat T Cells is Associated with Mitochondria-Dependent Caspase-3 Activation Regulated by Bcl-2

  • Park, Hye-Won;Jen, Do-Youn;Kim, Young-Ho
    • Journal of Microbiology and Biotechnology
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    • 제12권6호
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    • pp.950-956
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    • 2002
  • The antitumor activity of the insect pathogenic fungus Paecilomyces japonica has been attributed to apoptotic cell death. However, the mechanism underlying the induced apoptosis has not yet been elucidated. In this study, we for the first time show that mitochondria-dependent caspase-3 activation were associated with the apoptotic activity of P. japonica in human acute leukemia Jurkat T cells. When Jurkat T cells were treated with the ethyl acetate extract of P japonica at concentrations ranging from $2-6{\mu}g/ml$, apoptotic cell death. accompanied by several biochemical events such as caspase-9 activation, caspase-3 activation, degradation of poly (ADP-ribose) polymerase (PARP), and apoptotic DNA fragmentation, was induced in a dose-dependent manner. In addition, the release of cytochrome c from mitochondria was detected. Under these conditions, the expression of Fas and Fas-ligand (FasL) remained unchanged. Ethyl acetate extract-induced mitochondrial cytochrome c release, caspase-3 activation, PARP cleavage, and apoptotic DNA fragmentation were suppressed by the ectopic expression of Bcl-2, which is known to block mitochondrial cytochrorme c release. Accordingly, these results demonstrate that P. japonica-induced apoptotic cell death is mediated by a cytochrome c-dependent caspase-3 activation pathway that can be interrupted by Bcl-2.

계혈등(鷄血藤)의 Beta-sitosterol 성분이 자궁근종세포의 증식억제와 세포자멸사의 유도에 미치는 영향 (The effect of β-sitosterol proliferation and apoptosis in human uterine leiomyoma cells)

  • 박영선;백승희
    • 대한한방부인과학회지
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    • 제18권1호
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    • pp.181-191
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    • 2005
  • Purpose : ${\beta}$-sitosterol is kind of phytosterols or plant which are structurally similar to cholesterol. This study was aimed to investigate the inhibitory effect of the ${\beta}$-sitosterol on the proliferation of human uterine leiomyoma cells and the expression of gene related the mechanism of cell apoptosis. Methods : We counted the number of death cells treated with indicated time of the ${\beta}$-sitosterol and investigated cell death rate by cell count assay. Furthermore, flow cytometry analysis and DNA fragmentation assay were used to dissect between necrosis and apoptosis. and then we observed the differential gene expression by western blot analysis. Results : 1) The inhibitory effect on the growth of uterine leiomyoma cell treated with the ${\beta}$-sitosterol $16{\mu}M$ was increased in a time dependent. 2) The result of flow cytometry analysis, subG1 phase arrest related cell apoptosis was investigated 16.97% in uterine leiomyoma cell treated with the ${\beta}$-sitosterol $16{\mu}M$ and showed the fashion of proportional time dependent. 3) The gene expression of p27, p21 related cell cycle was increased according to increasing time interval but cyclin E-CDK2 complex was decreased expression. 4) The character of apoptosis, DNA fragmentation was significantly observed on the time dependent. 5) The expression of pro-caspase 3 and PARP were decreased dependent on treatment with time dependent. Conclusion : This study showed that the ${\beta}$-sitosterol have the inhibitory effect on the proliferation of human uterine leiomyoma cell and the effect was related with apoptosis.

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NMR and Fluorescence Studies of DNA Binding Domain of INI1/hSNF5

  • Lee, Dongju;Moon, Sunjin;Yun, Jihye;Kim, Eunhee;Cheong, Chaejoon;Lee, Weontae
    • Bulletin of the Korean Chemical Society
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    • 제35권9호
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    • pp.2753-2757
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    • 2014
  • INtegrase Interactor 1 protein (INI1/hSNF5) or BRG1-associated factor 47 (BAF47) is a SWI/SNF-related matrix associated actin dependent regulator of chromatin subfamily B member. DNA binding domain of INI1/hSNF5 is cloned into E.coli expression vectors, pET32a and purified as a monomer using size exclusion chromatography. NMR data show that $INI1^{DBD}$ has folded state with high population of ${\alpha}$-helices. By fluorescence-quenching experiments, binding affinities between $INI1^{DBD}$ and two double stranded DNA fragments were determined as $29.9{\pm}2.6{\mu}M$ (GAL4_1) and $258.7{\pm}5.8$ (GAL4_2) ${\mu}M$, respectively. Our data revealed that DNA binding domain of INI1/hSNF5 binds to transcriptional DNA sequences, and it could play an important role as a transcriptional regulator.

Ginkgo biloga 잎 추출물의 1,2,4-benzenetriol에 대한 항산화 효과에 대한 연구 (Protective Effects of Ginkgo Biloba Leaf Extract(GBE) against 1,2,4-benzenetriol Induced Toxicity in Vitro)

  • 이영준;김태연;정해원
    • 한국환경보건학회지
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    • 제27권1호
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    • pp.124-130
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    • 2001
  • Ginkgo biliba has been used for bronchitis and asthma in oriental countries and its leaf extract(GBE) contains 24% ginkgoflavone glycoside and 6% terpenoid. Flavonoids and terpenoids are known to have various antioxidant effects such as scavenging of free radicals and chelation of transtional metals. Antioxidant effect of GBE against 1,2,4-benzenetriol(BT), one of toxic metabolites of benzene, was demonstrated throughbsister chromatid exchange(SCE) analysis, single cell gel electrophoresis(SCGE) analysis, DNA cleavage assay and lipid peroxidation production analysis. The means of SCE frequencies at 10, 25 and 50$\mu$M concentration of BT were 7.72, 8.02, 9.22 respectively. In addition of GBE with concentration of 50, 200 and 500$\mu\textrm{g}$/$m\ell$, SCE frequencies were decreased significantly.(p<0.05) According to SCGE analysis, BT induced DNA damage in a dose-dependent manner at concentration of 10 and 50 $\mu$m and the DNA damage induced by BT was significantly protected by GBE(p<0.001). No genotoxicity was observed by GBE treatment alone on DNA cleavage. The effect of BT on lipid peroxidation product, Malondiadehyde(MDA), was increased with concentration of BT(10 and 50 $\mu$M) and reduction in MDA was noted when GBE was added. From above results it is suggested that GBE could protect the cell and DNA from pro-oxidant effect by reactive oxigen species induced by BT.

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Genes expression monitoring using cDNA microarray: Protocol and Application

  • Muramatsu Masa-aki
    • 한국독성학회:학술대회논문집
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    • 한국독성학회 2000년도 국제심포지움 및 추계학술대회
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    • pp.31-41
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    • 2000
  • The major issue in the post genome sequencing era is determination of gene expression patterns in variety of biological systems. A microarray system is a powerful technology for analyzing the expression profile of thousands of genes at one experiment. In this study, we constructed cDNA microarray which carries 2,304 cDNAS derived from oligo-capped mouse cDNA library. Using this hand-made microarray we determined gene expression in various biological systems. To determine tissue specific genes, we compared Nine genes were highly-expressed in adult mouse brain compared to kidney, liver, and skeletal muscle. Tissue distribution analysis using DNA microarray extracted 9 genes that were predominantly expressed in the brain. A database search showed that five of the 9 genes, MBP, SC1, HiAT3, S100 protein-beta, and SNAP25, were previously known to be expressed at high level in the brain and in the nervous system. One gene was highly sequence similar to rat S-Rex-s/human NSP-C, suggesting that the gene is a mouse homologue. The remaining three genes did not match to known genes in the GenBank/EMBL database, indicating that these are novel genes highly-expressed in the brain. Our DNA microarray was also used to detect differentiation specific genes, hormone dependent genes, and transcription-factor-induced genes. We conclude that DNA microarray is an excellent tool for identifying differentially expressed genes.

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DNA손상 및 돌연변이에 대한 명지버섯의 방어효능 (The Protective Effects of Ganoderma lucidum on the DNA Damage and Mutagenesis)

  • 이길수;공석경;최수영
    • Biomolecules & Therapeutics
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    • 제11권2호
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    • pp.139-144
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    • 2003
  • Ganoderma lucidum is commonly known as medically potent mushroom, which has been widely used in China and other oriental countries for the treatment of various diseases, including cancer. In this report, we investigated the anti-oxidant and protective effect of Ganodema lucidum extract (GLE) against the DNA damage induced by free radical and U.V. In the assay of cell growth inhibition, the inhibitory cell growth rate induced by hydroxyl radical was dose-dependently decreased by GLE. This results support that GLE has a detoxifying activity against cytotoxicity of hydroxyl radical in E. coli cell. GLE also protected ColE1 plasmid DNA damage in the concentration of 200$\mu\textrm{g}$ per reaction on the DNA fragmentation assay. The nuclear tailing by hydrogen peroxide in single cell gel electrophoresis(SCGE) was decreased by GLE in the concentration of 50$\mu\textrm{g}$/ml. These data indicate that Ganoderma lucidum has an anti-oxidative activity to hydrogen peroxide. The mutation rate after irradiation of U.V. was reduced by 50$\mu\textrm{g}$/ml GLE and total number of Rif (Rifampicin) resistant mutants was decreased in a concentration dependent manner when added the GLE exogenously in a culture media. According to the results, it is likely that GLE has not only an anti-oxidative activity to hydroxyl radical but also an anti-mutagenic activity to U.V. mutagenesis.

C-G 링커 어댑터 PCR을 이용한 지놈워킹 (C-G Linker Adaptor PCR Method for Genome Walking)

  • 서효석;이영기;전은영;이정헌
    • 한국연초학회지
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    • 제37권1호
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    • pp.25-33
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    • 2015
  • Genome walking is a par ticular application for identifying sequences of unknown genomic regions adjacent to a known region. Many genome walking methods based on polymerase chain reaction (PCR) are available. Even if earlier techniques suffer from low reproducibility, inefficiency, and non-specificity, improved strategies have been developed. In this study, we present an alternative strategy: the genomic DNA is digested with restriction enzymes. After cytosine overhangs at 5' ends, the fragments are ligated to linker adaptor s had guanine overhang at 3' ends. Then nested PCR is performed. The improvements in this strategy focus on two points. The first is the C tailing method using Pfu polymerase instead of the A tailing method based on nontemplate-dependent terminal transferase activity of Taq polymerase. Therefore unintended modification of target DNA can be prevented without A tailing error. The second point is the use of C/G-specific ligation had advantage in the ligation efficiency compared with A/T-specific ligation. Therefore, the C-G linker PCR method increases ligation efficiency between digested genomic DNA and adaptor DNA. As a result, the quantity of target DNA to amplify by PCR is enriched. We successfully used G-C linker PCR to retrieve flanking regions bordering the phophinothricin resistance gene in genetically modified tobacco (GMO).

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