• 대한의생명과학회지
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• 제2권2호
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• pp.257-265
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• 1996

토끼 바이러스성 출혈증의 원인체를 실험 토끼에 접종하여 증식을 유도하고 간장에서 hematoxylin & eosin 염 색 에서 조직학적 진단과 세포내 viral RNA의 소재를 결정하기 위해 post-unicryl 포매한 block의 절편을 사용하여 단 염색과 전자현미경적 in situ hybridization을 시도하였다. 토끼 출혈증 viral RNA의 보합 결합에 이용하는 probe는 4717에서 4800(84bases)까지 oligonucleotide를 5'말단에 biotin-CE phosphoramidite로 표지하여 사용하였다. 보합결합물의 증명은 신호 표지로서 antibiotin antibody-l0nm gold를 사용하였으며, hybridization이나 증명은 기존 protocol에서 약간의 변법을 사용하였다. 0.02% glutaraldehyde에서 고정하고 unicryl resin 포매한 표본, biotinylated oligonucleotide probe, antibiotin antibody-l0nm gold로 실험한 결과 증강된 신호를 얻을 수 있었다. 특히 전처리를 생략하므로써 실험 과정을 간단하게 하여 신속한 결과를 얻을 수가 있었다. 전자현미경 in situ hybridization을 통하여 토끼 출혈증 바이러스의 주요 표적은 간세포로 감염 세포의 세포질 내 미토콘드리아와 핵 사이에서 immune gold입자가 뚜렷하게 표지 됨으로서 viral RNA를 증명할 수 있었다.

• KSBB Journal
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• 제22권6호
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• pp.387-392
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• 2007

DNA 마이크로어레이는 바이오칩의 발전에서 가장 주목받으며 발전하고 있는 분야로서 이에 대한 연구가 점차 확장하고 있다. DNA나 RNA 등 유전자의 매우 느린 확산속도를 극복하기 위하여 마이크로플루딕 바이오칩이 DNA 마이크로어레이에 적용되는 최근의 학술적인 사례들을 연구, 비교하였다. DNA 마이크로어레이에 적용된 미세유체 바이오칩은 상당수가 효율적인 hybridization을 달성하기 위한 믹싱 시스템이 많이 보고되었으며, 이 총설에서는 그에 대한 분석을 수행하여 유전자 hybridization 강화를 이룬 시스템에 대한 최근 동향을 가늠할 수 있게 하였다. 특별히 PDMS를 이용한 마이크로 펌프의 적용 등, 앞으로의 미세유체 DNA 마이크로어레이 발전가능성과 모델링의 한계점 등을 정리 분석해 보았다.

• 대한전기학회논문지:전기물성ㆍ응용부문C
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• 제51권6호
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• pp.265-270
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• 2002

The determination of DNA hybridization reaction can apply the molecular biology research, clinic diagnostics, bioengineering, environment monitoring, food science and application area. So, the improvement of DNA hybridization detection method is very important for the determination of this hybridization reaction. Several molecular biological techniques require accurate predictions of matched versus mismatched hybridization thermodynamics, such as PCR, sequencing by hybridization, gene diagnostics and antisense oligonucleotide probes. In addition, recent developments of oligonucleotide chip arrays as means for biochemical assays and DNA sequencing requires accurate knowledge of hybridization thermodynamics and population ratios at matched and mismatched target sites. In this study, we report the characteristics of the probe and matched, mismatched target oligonucleotide hybridization reaction using thermodynamic method. Thermodynamic of 5 oligonucleotides with central and terminal mismatch sequences were obtained by measured UV-absorbance as a function of temperature. The data show that the nearest-neighbor base-pair model is adequate for predicting thermodynamics of oligonucleotides with average deviations for $\Delta$H$^{0}$ , $\Delta$S$^{0}$ , $\Delta$G$_{37}$ $^{0}$ and T$_{m}$, respectively.>$^{0}$ and T$_{m}$, respectively.

• Tuberculosis and Respiratory Diseases
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• 제51권6호
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• pp.517-529
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• 2001

연구배경 : Filter hybridization이나 solution hybridization 방법들은 Northern blot나 slot blot에 비하여 빠르며 재현성이 높고 소량의 RNA를 측정할 수 있으며 한꺼번에 많은 시료들을 동시에 측정하는것이 가능하다. 방 법 : 저자들은 쥐를 대상으로 하여 SP-C mRNA를 filter hybridization과 solution hybridization 방법들로 각각 정량측정하여 방법론에 따른 분자생물학적 정도관리에 대한 관찰을 하기 위하여 이 연구를 시행하였다. 결 과 : 1) Solution hybridization 방법에 의한 SP-C mRNA의 정량측정을 위한 RNA 검체물의 최소량은 3pg 이상이었다. 2) Solution hybridization에 쓰이는 SP-C sense mRN A input 양과 유전자 cpm과의 Y=6.46 X+244(X=SP-C mRNA 전사체 Y=cpm)였고, 양자간의 상관계수 r은 0.99로 매우 밀접한 상관성을 보였다. 3) Filter hybridization방법에 의한 SP-C mRNA의 정량측정을 위한 RNA검체물의 최소량은 0.1ng 이상이었다. 4) SP-C의 sense 전사체 0, 0.1, 0.5, 1.0, 2.5 및 5 ng에 대한 cpm과의 표준곡선은 Y=2541.6 X+252.7 (X=SP-C mRNA 전사체 Y=CPM) 이고 상관계수 r은 0.99로 매우 밀접한 상관성을 보였다. 결 론 : Solution hybridization 방법은 다른 방법에 비해 RNA input 양을 아주 소량 사용하면서도 주위에 높은 배후 방사성 산란을 보이는 비특이성 DNA가 있음에도 불구하고 특정 시료의 양이 한정되어 있는 상황에서 RNA를 분석 검토하는데 매우 유용한 연구방법이며, 뿐만아니라 filter hybridization 방법은 mRNA를 정량 측정하는데 있어서 신속하고 재현성이 높으며, 한꺼번에 많은 시료들을 시행할 수 있고, 사료의 mRNA를 정량측정 하는데 있어서 유용한 방법임을 알 수 있었다.

• 대한전기학회:학술대회논문집
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• 대한전기학회 2003년도 추계학술대회 논문집 전기물성,응용부문
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• pp.89-91
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• 2003

The determination of DNA hybridization reaction can apply the molecular biology research, clinic diagnostics, bioengineering, environment monitoring, food science and other application area. So, the improvement of DNA detection system is very important for the determination of this hybridization reaction. In this study, we report the characterization of the probe and target oligonucleotide hybridization reaction using the evanescent field microscopy. First, we have fabricated DNA chip microarray. The particles which were immobilized oligonucleotides were arranged by the random fluidic self-assembly on the pattern chips, using hydrophobic interaction. Second, we have detected DNA hybridization reaction using evanescent field microscopy. The 5'-biotinylated probe oligonucleotides were immobilized on the surface of DNA chip microarray and the hybridization reaction with the Rhodamine conjugated target oligonucleotide was excited fluorescence generated on the evanescent field microscopy. In the foundation of this result, we could be employed as the basis of a probe olidonucleotide, capable of detecting the target oligonucleotide and monitoring it in a large analyte concentration range and various mismatching condition.

• 생명과학회지
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• 제9권4호
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• pp.358-367
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• 1999

Thirty strains of methicillin resistant Staphylococcus aureus were obtained from the clinical isolates. In order to investigate the pursuit of the pathogens of nosocomial infection, these strains were studied for antibiotic sensitivity as well as its resistant pattern. Among the methods of hybridization which directly confirm the specific antibiotic resistant genes by means of the recently developed specific probe DNA, dot blot hybridization and southern blot hybridization were performed and these two methods were compared in their sensitivity and specificity. Strains that is sensitive to cephalothin to the subject of methicillin resistant Staphylococcus aureus were in 43%. Those that are sensitive to cefoperazone and cefuroxime were 26% and 23%, respectively. In case of MIC, MIC50 of cefoperazone was 8 $\mu\textrm{g}$/$m\ell$, and MIC90 was 128 $\mu\textrm{g}$/$m\ell$ to be the lowest. As the results of plasmid DNA electrophoresis, most of methicillin resistant Staphylococcus aureus strains had more than 4 plasmids. These plasmids digested by BamHI, methicillin resistant Staphylococcus aureus is distributed as 10 fragments with the size of 65 kb to 1.5 kb. Dot blot hybridization were performed to examine the existence of mecA gene to show the detection rate of 50%. Southern blot hybridization were done to see if DNA bands which amplify the activity of digoxigenium-labeled probe by PCR were actually PCR products of mecA gene and it showed the detection rate of 53%. It can be concluded that the southern blot hybridization seemed to be better in sensitivity and specificity when it is compared with the results of dot blot hybridization.

• 한국실내디자인학회논문집
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• 제23권4호
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• pp.129-139
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• 2014

The objective of this research is to suggest new geometric possibilities in digital architecture by investigating the characteristics of hybridization in digital geometry. The research begins with theoretical background research such as defining hybridization, investigating hybrid thinking, and studying the theory of digital geometry, along with the four conceptual characteristics of hybridization that could be drawn, such as temporality, liquidity, complexity, and connectivity. Based on these characteristics, the generative method of hybrid digital geometric languages such as Blob, Particle, Morph, Loft, and Boolean was analyzed with case research in contemporary digital architecture. As a result, diverse hybrid geometric keywords were extracted; these keywords suggest potential meanings of hybridization such as accidentality, mobility, diversity, and identity. Different elements represent the "mobility" in time by the force and wave, and they are "accidentally" combined in gradual change. The united species in "diverse" characters are seamlessly connected and emerge as a new "identity." The research maximizes the generative possibilities in digital geometry and provides a theoretical basis to apply the digital hybrid methods to architectural design by suggesting the potential meanings and possibilities in hybridization.

• Journal of Microbiology and Biotechnology
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• 제4권3호
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• pp.235-236
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• 1994

Direct colony hybridization on agarose gel plate was established for the identification of recombinant plasmids. The hybridization of the probe to nucleic acids on dried gel without transferring to solid supports was more effective and simpler than hybridization of such probes to materials immobilized on filters such as nitrocellulose or nylon. D-cycloserine in overlaying agamse was essential for releasing the nucleic acids from colony.

• Journal of Microbiology and Biotechnology
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• 제17권6호
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• pp.945-951
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• 2007

DNA-DNA hybridization has been established as an important technology in bacterial species taxonomy and phylogenetic analysis. In this study, we analyzed how the efficiency with which the genomic DNA from one species hybridizes to the genomic DNA of another species (DNA-DNA hybridization) in microarray analysis relates to the similarity between two genomes. We found that the predicted DNA-DNA hybridization based on genome sequence similarity correlated well with the experimentally determined microarray hybridization. Between closely related strains, significant numbers of highly divergent genes (>55% identity) and/or the accumulation of mismatches between conserved genes lowered the DNA-DNA hybridization signal, and this reduced the hybridization signals to below 70% for even bacterial strains with over 97% 16S rRNA gene identity. In addition, our results also suggest that a DNA-DNA hybridization signal intensity of over 40% indicates that two genomes at least shared 30% conserved genes (>60% gene identity). This study may expand our knowledge of DNA-DNA hybridization based on genomic sequence similarity comparison and further provide insights for bacterial phylogeny analyses.

• International Journal of Industrial Entomology and Biomaterials
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• 제18권1호
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• pp.41-48
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• 2009

The large bumblebee, Bombus terrestris, indigenous to Europe and used extensively for high-value crop pollination, has been artificially introduced in several parts of the world. Here we show the interspecific hybridization between bumblebee species, B. terrestris and B. ignitus, under laboratory conditions. The mating and oviposition percentages of the interspecific hybridization of a B. terrestris queen with a B. ignitus male were higher than those of the intraspecific mating of B. ignitus. Furthermore, the competitive copulation experiment indicated that the mating of B. ignitus males with B. terrestris queens was 1.8-fold more frequent than with B. ignitus queens. The interspecific hybridization of a B. ignitus queen with a B. terrestris male produced either B. ignitus workers or the B. ignitus male phenotype, and the hybridization of a B. terrestris queen with a B. ignitus male produced B. terrestris males. Genetic tests using a portion of the mitochondrial COI gene for the parent and hybrid phenotypes indicated that mitochondrial DNA in the interspecific hybridization was maternally inherited. Our results indicated that interspecific hybridization occurred between B. ignitus and B. terrestris, which suggests that the hybridization will have a negative impact of competition and genetic pollution of native bumblebees.