• Title/Summary/Keyword: human cDNA

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Construction of Human cDNA Library Analysis Pipeline (인간 cDNA 라이브러리 분석 파이프라인 구축)

  • Jung, Jaeeun;Kim, Dae-Soo
    • Proceedings of the Korea Contents Association Conference
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    • 2018.05a
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    • pp.323-324
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    • 2018
  • 전장 cDNA 클론을 시퀀싱하는 것은 선택적 스플라이싱 형태를 비롯한 정확한 유전자 구조를 정의하는데 유용하게 사용될 수 있으며, 유전자 및 단백질의 생물학적 기능연구에 중요한 자원을 제공한다. 포괄적이며 비 중복적인 cDNA의 생산은 인간 유전체 연구의 중요한 목표이다. 본 연구에서 제공하는 인간 cDNA 라이브러리 분석 파이프라인은 전장 cDNA를 분석하는 자동화 도구로 여러 연구자들에게 활용 될 수 있을 것으로 사료된다.

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Construction of Human cDNA Library Analysis Pipeline (인간 cDNA 라이브러리 분석 파이프라인 구축)

  • Jung, Jaeeun;Kim, Dae-Soo
    • Proceedings of the Korea Contents Association Conference
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    • 2018.05a
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    • pp.83-84
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    • 2018
  • 전장 cDNA 클론을 시퀀싱하는 것은 선택적 스플라이싱 형태를 비롯한 정확한 유전자 구조를 정의하는데 유용하게 사용될 수 있으며, 유전자 및 단백질의 생물학적 기능연구에 중요한 자원을 제공한다. 포괄적이며 비 중복적인 cDNA의 생산은 인간 유전체 연구의 중요한 목표이다. 본 연구에서 제공하는 인간 cDNA 라이브러리 분석 파이프라인은 전장 cDNA를 분석하는 자동화 도구로 여러 연구자들에게 활용 될 수 있을 것으로 사료된다.

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A Plausible Method for the Diagnosis of Genetic Disorders Using Full Length cDNA

  • Hur, Hyang-Suk;Lee, Young-Won;Park, Hyoung-Woo;Kim, Myoung-Hee
    • Biomedical Science Letters
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    • v.7 no.1
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    • pp.1-5
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    • 2001
  • A cDNA of coagulation Factor IX gene has been screened from the $\lambda$gt11 human fetal liver cDNA library, and used to construct a 2.8-kb full length cDNA after recombining with the N-terminal fragment from pTZ-FIX. Human genomic DNA was isolated, digested with the restriction endonucleases, TaqI, EcoRI, and HindIII, and Southern hybridization was performed using the full length factor IX cDNA as a probe. The hybridized bands generated by the restriction endonucleases were the followings: TaqI, 0.3, 1.0, 1.6, 1.8, 2.7, 3.7, and 5.3 kb bands; EcoRI, 1.8, 4.8, 4.9, 5.5, 6.8, and 12.6 kb bands; HindIII, 4.1, 4.4, 5.2, 5.8, 7.6, and 12.5 kb bands. When the Southern bands were physically mapped along the genome, about 50-kb continuous region harboring almost all of the genomic region of Factor Ⅸ gene was covered. These results suggest a possibility of using an exonal cDNA probe to diagnose abnormalities including large deletions, insertions, and rearrangements along the genome, if there is any.

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Comparison of the Protective Effect of Antioxidant Vitamins and Fruits or Vegetable Juices on DNA Damage in Human Lymphocyte Cells Using the Comet Assay (Comet Assay를 이용한 항산화 비타민과 과일.야채즙의 인체 임파구 세포 DNA 손상 감소 효과 비교)

  • 전은재;박유경;김정신;강명희
    • Journal of Nutrition and Health
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    • v.37 no.6
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    • pp.440-447
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    • 2004
  • In this study the in vitro protective effects of several antioxidant vitamins (vitamin C, $\alpha$-tocopherol, $\beta$-carotene), fruits and vegetables (strawberry, tangerine, orange and 100% orange juice, carrot juice), on the levels of isolated human lymphocyte DNA damage was measured using Comet assay. Comet assay has been used widely to assess the level of the DNA damage in the individual cells. Lymphocytes were pre-treated for 30 minutes with antioxidant vitamins (10, 50, 100, 500 $\mu$M) or fruits$.$vegetables (10, 100, 500, 1000 $\mu$g/ml), an4 then oxidatively challenged with 100 $\mu$M $H_2O$$_2$ for 5 min at 4$^{\circ}C$. The protective effect of antioxidant vitamins against DNA damage at a concentration of 50 $\mu$M were 50% in vitamin C, 32% in $\alpha$-tocopherol, whereas, fJ-carotene showed a 55% protection at a dose as low as 10 $\mu$M. The inhibitory effects of DNA damage by strawberry, tangerine, orange, orange juices, carrot juices were 50 - 60% with wide ranges of doses. The results of the present study indicate that most the antioxidant vitamins and fruits$.$vegetables juices produced a significant reduction in oxidative DNA damage.

Chemopreventive Effect of Quercetin, Vitamin C and Trolox Against the Organic Extract of Airborne Particulate Matter Induced Genotoxicity in A549 Human Lung Carcinoma Cells (대기부유분진추출물로 야기된 DNA 손상에 대한 Quercetin, Vitamin C 및 Trolox 의 보호효과)

  • Kim, Nam-Yee;Heo, Moon-Young
    • YAKHAK HOEJI
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    • v.51 no.4
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    • pp.239-245
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    • 2007
  • In order to evaluate the genotoxicity of airborne particulate matter extracted with dichloromethane (APE), the rat microsome mediated (S-9) or DNA repair enzyme treated Comet assays were performed using the single cell gel electrophoresis in A549 human lung carcinoma cells. It was found that the cells interacting with APE showed more DNA single-strand breaks relative to untreated cells. The genotoxicity of APE was increased with the treatment of S-9 mixture. Microsome mediated DNA damage was inhibited by CYP1Al inhibitor, quercetin. The APE also showed oxidative DNA damage evaluated by endonuclease III treatment. Oxidative DNA damage of APE was inhibited by antioxidants such as vita- min C and Trolox. We also found that the vegetables or fruits extract may reduce APE-induced genotoxicity by their anti- oxidant activity and CYP1A1 inhibition.

Transcriptional Responses of Human Respiratory Epithelial Cells to Nontypeable Haemophilus influenzae Infection Analyzed by High Density cDNA Microarrays

  • Lee, Ji-Yeon;Lee, Na-Gyong
    • Journal of Microbiology and Biotechnology
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    • v.14 no.4
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    • pp.836-843
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    • 2004
  • Nontypeable H. influenzae (NTHi), a Gram-negative obligate human pathogen, causes pneumonia, chronic bronchitis, and otitis media, and the respiratory epithelium is the first line of defense that copes with the pathogen. In an effort to identify transcriptional responses of human respiratory epithelial cells to infection with NTHi, we examined its differential gene expression using high density cDNA microarrays. BEAS-2B human bronchial epithelial cells were exposed to NTHi for 3 hand 24 h, and the alteration of mRNA expression was analyzed using microarrays consisting of 8,170 human cDNA clones. The results indicated that approximately 2.6% of the genes present on the microarrays increased in expression over 2-fold and 3.8% of the genes decreased during the 24-h infection period. Upregulated genes included cytokines (granulocyte-macrophage colony stimulating factor 2, granulocyte chemotactic protein 2, IL-6, IL-10, IL-8), transcription factors (Kruppel-like factor 7, CCAAT/enhancer binding protein $\beta$, E2F-1, NF-$\kappa$B, cell surface molecules (CD74, ICAM-1, ICAM-2, HLA class I), as well as those involved in signal transduction and cellular transport. Selected genes were further confirmed by reverse-transcription-PCR. These data expand our knowledge of host cellular responses during NTHi infection and should provide a molecular basis for the study of host-NTHi interaction.

Identification of Bak-like Protein cDNA (Bak-like 단백질을 code하는 cDNA의 동정)

  • 김진경
    • YAKHAK HOEJI
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    • v.45 no.4
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    • pp.426-430
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    • 2001
  • Cells are eliminated in a variety of physiological settings by apoptosis, a genetically encoded process of cellular suicide. Bak, a member of the Bcl-2 protein family, accelerates apoptosis by an unknown mechanism. We have found a novel cDNA encoding a 101 amino acid protein possessing a Bak-like in our full-length cDNA bank. Bak-like shares the conserved domains BHI and 2 with other proapoptotic proteins but lacks the BH3 domain. Bak-like is expressed in a wide variety of tissues. Like Bak, Bak-like gene product primarily enhances apoptotic cell death following an appropriate stimulus.

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Effects of Storage Buffer and Temperature on the Integrity of Human DNA

  • Kim, Yun-Tae;Choi, Eun-Hee;Son, Bo-Kyoung;Seo, Eun-Hee;Lee, Eun-Kyoung;Ryu, Je-Kwon;Ha, Gi-Won;Kim, Jin-Seon;Kwon, Mi-Ran;Nam, Jae-Hoon;Kim, Young-Jin;Lee, Kyoung-Ryul
    • Korean Journal of Clinical Laboratory Science
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    • v.44 no.1
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    • pp.24-30
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    • 2012
  • In this study, we have examined the effects of the storage time and temperature on DNA quality and have also studied the effects of the hydration buffer in which DNA is dissolved. This study was performed using 160 human blood samples collected with informed consent from 2007 to 2008 in the hospital where this cohort study was performed. The DNA extracted was dissolved using distilled water (DW) or Tris-EDTA (TE) buffer, and stored in the deep freezer or refrigerator for up to 10 weeks at $-70^{\circ}C$, $-20^{\circ}C$, $4^{\circ}C$, and $25^{\circ}C$, respectively. DNA integrity was determined by the degree of smearing of DNA on the gel. After four weeks, all of the 20 DNA samples dissolved in DW and stored at $25^{\circ}C$ were entirely degraded. After 10 weeks, 6 of the 20 DNA samples dissolved in TE buffer and stored at $25^{\circ}C$ were fairly degraded, and 4 of the 20 DNA samples dissolved in DW and stored at $4^{\circ}C$ were fairly degraded. The 20 DNA samples dissolved in TE buffer and stored at $4^{\circ}C$ were stable for 10 weeks. DNA samples stored at $-20^{\circ}C$ and $-70^{\circ}C$ did not appear to degrade in either DW or TE buffer, even at the 10-week point. We suggest that TE buffer should use for DNA elution, in order to protect against degradation and to preserve DNA for a long period of time, and the samples should be stored at $-20^{\circ}C$ or $-70^{\circ}C$.

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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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Protective Effect of Electrolyzed Reduced Water on the Paraquat-induced Oxidative Damage of Human Lymphocyte DNA (Paraquat에 의한 사람 임파구 DNA 손상에 대한 환원전리수의 보호효과)

  • Park, Eun-Ju;Ryoo, Kun-Kul;Lee, Yoon-Bae;Lee, Jong-Kwon;Lee, Mi-Young
    • Applied Biological Chemistry
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    • v.48 no.2
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    • pp.155-160
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    • 2005
  • Electrolyzed reduced water (ERW), showing extremely negative oxidation-reduction potential, was used to investigate the effects of paraquat-induced damages on DNA from human lymphocyte. The effect of ERW on paraquat-induced oxidative DNA damage in human lymphocytes was evaluated by Comet assay (single-cell gel electrophoresis) quantified as percentage fluorescence in tail. Comet assay has been used widely to assess the level of the DNA damage in individual cells. Lymphocytes were oxidatively challenged with various concentrations of paraquat for 30 min at $37^{\circ}C$, and were then treated with electrolyzed reduced water for 30 min. The oxidative DNA damage by paraquat, as indicated by the fluorescent tail in DNA, increased in a dose-dependent manner. However, oxidative damage of the DNA was almost completely prevented upon treatment with electrolyzed reduced water.