• Title/Summary/Keyword: DNA 칩

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Technology and Application of Cells on Chips (세포칩 기술과 응용)

  • Kim, C.B.;Song, K.B.
    • Electronics and Telecommunications Trends
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    • v.26 no.3
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    • pp.95-104
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    • 2011
  • 셀칩(cells on chips)이란, MEMs/NEMs 응용분야 중 생명공학과 관련된 세포분야로의 응용에 이용되는 대표적인 기술로서 현재 전세계에서 경쟁적으로 연구, 개발되고 있다. 셀칩은 생체내부에서 세포가 성장하는 공간적(spatial), 시간적(temporal) 조건을 정교하게 모사(mimicking)함으로써, 복잡한 생화학적 생체 내(in vivo) 환경을 이해할 수 있는 새로운 기회를 창조하고 있다. 또한 셀칩과 다양한 형태의 분석용 센서와의 결합된 시스템을 통하여, 세포기반 질병진단 시스템의 소형화 및 조기진단 시스템 개발을 위한 바이오멤스 핵심 플랫폼 기술로 인식되고 있다. 즉 DNA, 단백질, 세포 등의 바이오 물질을 마이크로/나노시스템 위에서 검출 및 분석함으로써 극미량의 생체물질을 실시간 고감도 분석이 가능하게 할 것이다. 본 고에서는 셀칩분야의 기술 및 응용에 관해 정리하고 있다.

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Fluorescece Microscope using Total Internal Reflection for Measuring Biochip (내부 전반사 방식에 의한 바이오칩 측정 장비)

  • Bae, Soo-Jin;Kang, Uk
    • The Transactions of The Korean Institute of Electrical Engineers
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    • v.56 no.9
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    • pp.1694-1698
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    • 2007
  • This study suggests a new fluorescence microscope to observe micro-samples within fluorophore in a variety of biomedical fields including the fluorescence analysis of a biochip, such as a DNA micro-array. A fluorescence microscope is a device for irradiating light onto a micro-object, executing an excitation and fluorescence emission process. In this study, it adopts a total internal reflection fluorescence(TIRF) method to excite a whole micro-sample substrate different from an existing way which uses an evanescent wave resulting from a total internal reflection on the micro-sample surface. Suggested TIRF microscope can reduce optical noise and obtain images with higher sensitivity thus obtain precise information about the density, quantity, location, etc. of a flurophore, and can simultaneously process separate images even when plurality of fluorophores having different excitation and fluorescent wavelength ranges is distributed, thus easily obtain information about the fluorophores.

Dimension Reduction in Time-series Gene Expression Data using incremental PCA (점진적 주성분 분석을 이용한 시계열 유전자 발현 데이터의 효율적인 차원 축소)

  • Kim, Sun-Hee;Kim, Man-Sun;Yang, Hyung-Jeong
    • Proceedings of the Korea Information Processing Society Conference
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    • 2007.11a
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    • pp.733-736
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    • 2007
  • 최근 생명 공학 기술의 발달로 마이크로 단위의 실험이 가능해지고 하나의 칩상에 수 만개의 유전자들의 발현 양상을 보다 쉽게 관찰할 수 있게 되었다. DNA 칩 기술에 의해 얻어지는 마이크로어레이(microarray) 데이터는 세포나 조직 내의 유전자 발현도(expression level)를 측정한 것으로 질병 진단이나 유전자 기능 예측 등에 이용되고 있다. 본 논문에서는 대량의 시계열 마이크로어레이 데이터 분석을 위해 효율적으로 데이터의 차원을 판단하는 점진적 주성분 분석을 이용하여 데이터의 차원을 축소 한다. 제안된 방법은 실제 시계열 마이크로어레이 데이터인 yeast cell cycle 데이터에 적용되었고, 데이터 차원 축소에 대한 효율성을 검증하기 위해 클러스터링을 수행하였다. 그 결과 데이터를 축소하여 클러스터링을 수행한 경우 학습 성능이 향상 된 결과를 보였다.

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Microarray Analysis of Gene Expression in Rat Hippocampus of Maternal Social Separation Model (모성 및 사회성 분리 백서 모델의 해마에서 유전자 칩을 이용한 유전자 발현 연구)

  • Lee, Hee Jae;Son, Chang Hee;Kwak, Hyong Ryol;Lee, Sang-Hyun;Han, Yoon Hee;Kim, Soo Young;Park, Jong-Ik;Chun, Wanjoo;Kim, Sung-Soo
    • Korean Journal of Biological Psychiatry
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    • v.13 no.2
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    • pp.110-116
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    • 2006
  • Objectives : Alteration of hippocampus was demonstrated in the maternal social separation(MSS) pups, separated from dams on postnatal day(pnd) 14 and placed alone. Therefore, to understand the molecular events involved in the MSS, we have initiated a search for gene profiles that are up or down-regulated in the hippocampus of MSS pups. Methods : Analysis of cDNA microarray was performed by using total RNA extracted from the hippocampus of control and MSS pups on pnd 17. Also, passive-avoidance test was demonstrated on pnd 35. Results : Up-regulation of Nedd4a was observed in the hippocampus of MSS pups. Also, MSS rats showed less elongation of latency in passive avoidance test. Conclusion : We suggest that environmental effects of MSS may be altered the neural and/or glial differentiation and synapse formation-related genes which may lead cognitive alterations in MSS rats.

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자기조립박막 증착법을 이용한 영구적인 친수성 표면의 플루이딕스칩 제작에 관한 연구

  • Kim, Dong-Jin;Lee, Mun-Gwon;Lee, Jeong-Hwan;Im, Hyeon-U;Park, Jin-Gu;Sin, Sang-Taek;Kim, Jeong-Ho;Jo, Byeong-Gi
    • Proceedings of the Materials Research Society of Korea Conference
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    • 2009.05a
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    • pp.49.2-49.2
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    • 2009
  • 최근, 플루이딕스칩 제작에 있어서 가격이 저렴하며 구조물 형성이 쉽다는 장점으로 인하여 유리 기판을 플라스틱 기판으로 대체하려는 연구가 많이 진행하고 있다. 하지만 플라스틱 기판은 유리 기판에 비하여 많은 장점을 갖고 있음에도 불구하고, 기판 표면이 소수성이기 때문에 유체의 흐름을 저하시키는 문제점이 있다. 기존의 플라스틱 기판을 친수성으로 개질하는 방법으로는 화학적처리, 자외선 조사, 산소플라즈마 처리 등의 방법이 있었으나, 화학적처리 방법은 공정의 민감성과 폐기물로 인한 양산적용의 한계가 있고, 자외선 조사법 및 산소 플라즈마 처리는 친수성이 영구적이지 않다는 결정적인 문제점이 있다. 이는 플라스틱 플루이딕스칩의 신뢰도를 크게 저해하여 상용화에 큰 문제점으로 작용한다. 이러한 문제점을 극복하기 위한 새로운 방법의 친수성 표면처리가 요구되어 지고 있다. 본 연구에서는, 기존 플라스틱 기판의 친수성 표면처리 방법들의 문제점들을 개선하고자 플라스틱 기판의 변형을 야기하지 않는 저온 PE-CVD 방식을 이용하여 균질한 두께의 $SiO_2$박막을 형성 하였으며, 형성된 박막을 liquid self-assembled monolayer(L-SAM)방식을 이용하여 아민 표면으로 개질하였다. 이를 통해, 플라스틱 채널 상에서 유체의 원활한 흐름, 형성된 아민 표면에 단백질 및 DNA와 같은 생체 물질 고정화의 용이성 및 영구적 표면개질의 특성을 얻을 수 있었다. 이뿐만 아니라, 플라스틱 기판 외에도 재료에 관계없이 모든 물질의 표면을 생체 안정성이 뛰어난 친수성 표면으로 개질 할 수 있으며, 알데히드 및 카르복시산 등의 다양한 작용기로 변형이 쉽다는 장점이 있다. 개질된 친수성 표면의 평가를 위하여 시간에 따른 접 촉각 및 형광 스캐너(Fluorescence Scanner)를 이용하여 영구적인 친수성 특성 및 생체물질 적합성을 파악하였다. 또한, L-SAM 조건에 따른 아민의 형성정도를 측정하기 위하여 XPS(X-ray Photoelectron Spectroscopy) 분석을 실시 하였다. 최적화된 표면처리를 실제 플라스틱 플루이딕스칩에 적용하여, 유체의 흐름을 관찰 하였다.

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Imputation Accuracy from 770K SNP Chips to Next Generation Sequencing Data in a Hanwoo (Korean Native Cattle) Population using Minimac3 and Beagle (Minimac3와 Beagle 프로그램을 이용한 한우 770K chip 데이터에서 차세대 염기서열분석 데이터로의 결측치 대치의 정확도 분석)

  • An, Na-Rae;Son, Ju-Hwan;Park, Jong-Eun;Chai, Han-Ha;Jang, Gul-Won;Lim, Dajeong
    • Journal of Life Science
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    • v.28 no.11
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    • pp.1255-1261
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    • 2018
  • Whole genome analysis have been made possible with the development of DNA sequencing technologies and discovery of many single nucleotide polymorphisms (SNPs). Large number of SNP can be analyzed with SNP chips, since SNPs of human as well as livestock genomes are available. Among the various missing nucleotide imputation programs, Minimac3 software is suggested to be highly accurate, with a simplified workflow and relatively fast. In the present study, we used Minimac3 program to perform genomic missing value substitution 1,226 animals 770K SNP chip and imputing missing SNPs with next generation sequencing data from 311 animals. The accuracy on each chromosome was about 94~96%, and individual sample accuracy was about 92~98%. After imputation of the genotypes, SNPs with R Square ($R^2$) values for three conditions were 0.4, 0.6, and 0.8 and the percentage of SNPs were 91%, 84%, and 70% respectively. The differences in the Minor Allele Frequency gave $R^2$ values corresponding to seven intervals (0, 0.025), (0.025, 0.05), (0.05, 0.1), (0.1, 0.2), (0.2, 0.3). (0.3, 0.4) and (0.4, 0.5) of 64~88%. The total analysis time was about 12 hr. In future SNP chip studies, as the size and complexity of the genomic datasets increase, we expect that genomic imputation using Minimac3 can improve the reliability of chip data for Hanwoo discrimination.

Detection for Methicillin Resistant Staphylococcus aureus in Using Bio-Chip Based Loop Mediated Isothermal Amplification Assay (칩 기반 등온증폭법을 이용한 약제 내성 포도상구균의 검출)

  • Cho, Min-Ho;Jang, Won-Cheoul;Choi, Jae-Gu
    • Journal of the Korean Chemical Society
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    • v.57 no.1
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    • pp.81-87
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    • 2013
  • Staphylococcus aureus is the most important pathogen in nosocomial infections, including bloodstream infections. Prompt identification of S. aureus from blood cultures and detection of methicillin resistance are essential in cases of suspected sepsis. We have studied a new method for the sequence-specific visual detection of minute amounts of nucleic acids using intercalating reaction by addition of SYBR Green to amplicons of LAMP, and it's a unique gene amplification method in which DNA can be isothermally amplified using only one enzyme. Staphylococcus-LAMP, which targets the spa gene, encoding S.aureus-specific protein A, and the mecA gene, encoding penicillin-binding protein-2' for methicillin resistance, detected MRSA and MRSE. In this study, by using LAMP assay, I detected for Staphylococcus aureus and Staphylococcus epidermidis concentration in the clinical sample. The detection of Staphylococcus aureus and Staphylococcus epidermidis was tested by using serial 10-fold dilutions standard solution. I have accurate detected the limit of detection, sensitity, specificity and reproducibility of the assay. The Bio-chip based LAMP assay allowed easy, rapid, accurate and sensitive detection of infection with Staphylococcus and especially applicable in a resource-limited situation.

Prevalence of Human Papillomavirus by DNA Chip Test in Women (여성에 있어 DNA 칩검사에 의한 인유두종바이러스 감염률의 조사)

  • Kim, Jae-Woo;Kim, Yun-Tae;Kim, Dae-Sik;Choi, Seok-Cheol
    • Journal of Life Science
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    • v.18 no.12
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    • pp.1657-1664
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    • 2008
  • We determined the prevalence of human papillomavirus (HPV) by DNA chip test in 549 women and cytologic diagnosis. 237 of 549 women (43.17%) subjected with HPV DNA Chip examination were found positive for HPV. 210 (88.60%, High group) were infected with high-risk HPV types. 17 (7.17%, Low group) were infected with low-risk HPV types (6, 11, 40, 44, 70) and 17 (7.17%, Mixed group) were infected with mixed types. According to age, in their twenties, thirties, forties, fifties and over sixties, the prevalence of infection with high-risk HPV types were 1.26% (3/237), 15.61% (37/237), 31.65% (75/237), 23.21% (55/237), and 13.92% (33/237), respectively. In the Low and Mixed group, percentages of infection with HPV were significantly lower than that of the High group. On the comparison of cytologic diagnosis (224 women) by Pap smear and DNA chip positive (237 women) for HPV, 132 out of 194 cases in the High group (68.04%) suffered cervical lesions with ASCUS (atypical squamous cells of undetermined significance, 7.22%), LSIL (low grade squamous intraepithelial lesion, 15.98%), HSIL (high grade SIL, 23.20%) and ICC (invasive cervical cancer, 21.65%). The Low group (14/224 women) showed 1 case of ASCUS and 6 cases of LSIL, whereas the Mixed group (4/224 women) had only 2 cases of ASCUS. According to the HPV subtypes, the high-risk types 16 and 18 induced 26 and 7 cases of ICC, respectively, whereas other HPV subtypes induced lower or no ICC incidence. In conclusion, the present data imply that the prevalence of HPV was 43.17%, high-risk HPV type 16 is a major factor, which causes precancerous and/or cervical cancer in woman and that HPV DNA chip is an accurate and useful tool for detecting HPV.

Isolation and Functional Identification of BrDSR, a New Gene Related to Drought Tolerance Derived from Brassica rapa (배추 유래 신규 건조 저항성 관련 유전자, BrDSR의 분리 및 기능 검정)

  • Yu, Jae-Gyeong;Park, Young-Doo
    • Horticultural Science & Technology
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    • v.33 no.4
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    • pp.575-584
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    • 2015
  • Drought stress is a crucial environmental factor determining crop survival and productivity. The goal of this study was to clearly identify a new drought stress-tolerance gene in Brassica rapa. From KBGP-24K microarray data with the B. rapa ssp. pekinensis inbred line 'Chiifu' under drought stress treatment, a gene which was named BrDSR (B. rapa Drought Stress Resistance) was chosen among 738 drought-responsive unigenes. BrDSR function has yet to be determined, but its expression was induced over 6-fold by drought. To characterize BrDSR, the gene was isolated from B. rapa inbred line 'CT001' and found to contain a 438-bp open reading frame encoding a 145 amino acid protein. The full-length cDNA of BrDSR was used to construct an over-expression vector, 'pSL100'. Tobacco transformation was then conducted to analyze whether the BrDSR gene can increase drought tolerance in plants. The BrDSR expression level in T1 transgenic tobacco plants selected via PCR and DNA blot analyses was up to 2.6-fold higher than non-transgenic tobacco. Analysis of phenotype clearly showed that BrDSR-expressing tobacco plants exhibited more tolerance than wild type under 10 d drought stress. Taking all of these findings together, we expect that BrDSR functions effectively in plant growth and survival of drought stress conditions.

Characterization of a Drought-Tolerance Gene, BrDSR, in Chinese Cabbage (배추의 건조 저항성 유전자, BrDSR의 기능 검정)

  • Yu, Jae-Gyeong;Lee, Gi-Ho;Park, Young-Doo
    • Horticultural Science & Technology
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    • v.34 no.1
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    • pp.102-111
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    • 2016
  • The goal of this study was to characterize the BrDSR (Drought Stress Resistance in B. rapa) gene and to identify the expression network of drought-inducible genes in Chinese cabbage under drought stress. Agrobacterium-mediated transformation was conducted using a B. rapa inbred line ('CT001') and the pSL100 vector containing the BrDSR full length CDS (438 bp open reading frame). Four transgenic plants were selected by PCR and the expression level of BrDSR was approximately 1.9-3.4-fold greater than that in the wild-type control under drought stress. Phenotypic characteristics showed that BrDSR over-expressing plants were resistant to drought stress and showed normal growth habit. To construct a co-expression network of drought-responsive genes, B. rapa 135K cDNA microarray data was analyzed to identify genes associated with BrDSR. BrDSR was directly linked to DARK INDUCIBLE 2 (DIN2, AT3G60140) and AUTOPHAGY 8H (ATG8H, AT3G06420) previously reported to be leaf senescence and autophagy-related genes in plants. Taken together, the results of this study indicated that BrDSR plays a significant role in enhancement of tolerance to drought conditions.