• Title/Summary/Keyword: hybridization

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Spermatogonia 단계에 특이적으로 발현하는 유전자 동정

  • 옥도원;김진회
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.48-48
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    • 2003
  • 본 실험은 spermatogonia 단계에 발현하는 유전자를 찾기 위하여 suppression subtractive hybridization를 수행하였다. 기존에 mouse에서는 spermatogonia 특이적인 유전자들이 밝혀져 있기 때문에 pig에 특이적인 유전자를 찾기 위하여 pig 250days testis와 pig 60days testis를 재료로 하여 실험하였다. SSH를 통하여 254days testis에 특이적으로 발현되는 후보유전자를 7개 찾았고 25days testis와 60days testis 의 Northern blot을 통하여 25days에 과발현하고 60days에 발현의 양이 대폭 줄어드는 spermatogonia 유전자로 생각되는 후보유전자 2개를 선택하여 pig tissue northern blot, genomic DNA southern blot, RT-PCR 그리고 In-situ hybridization을 수행하였다. Tissue northern blot과 RT-PCR을 통하여 후보자 1번은 간과 폐, 난소, 정소에서 발현하고, 후보유전자 15번은 난소와 정소에서만 특이적으로 발현함을 알았다. DNA sequence analysis와 NCBI Blast search를 통하여 후보자 1번은 다른 종에서 밝혀진 유전자였고 후보유전자 15번은 어느 종에서도 밝혀지지 않은 새로운 유전자였다. Degenerated primer를 통하여 후보자 1번의 pig full sequence를 밝히고 NCBI에 등록하였다. 그리고 In-situ hybridization을 통하여 후보유전자득이 20일째 testis의 Leydic cell에서 많이 발현되고 adult testis에서는 발현이 감소하는 결과를 얻었다. 이것으로 보아 위의 두 후보유전자는 spermatogonia에 직접 관련된 유전자이기 보다는 spermatogonia의 발달에 영향을 주는 leydic cell 특이발현을 가진 유전자로 사료되어진다.

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Utilization of F2 Seeds in Forest Tree Breeding (임목(林木)에서의 F2 이용(利用))

  • Jhun, Gae Sang;Sohn, Sung In
    • Journal of Korean Society of Forest Science
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    • v.70 no.1
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    • pp.109-114
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    • 1985
  • Utilization of $F_2$ seeds in forest tree hybridization program could be as dangerous as it could turn out to be handy and beneficial. Variation in $F_2$ generation does not appear to be dangerous as many breeder's concern. Pitch-loblolly pine hybridization program in Korea is reviewed and use of $F_2$ seeds are strongly recommended under the strict guide line of further breeding program. Breeding strategy for pitch-loblolly pine hybridization is suggested.

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Pathological findings and virus detection by in situ hybridization in the Korean native goats experimentally infected with Aujeszky's disease virus (오제스키병바이러스 인공감염 한국재래산양의 병리학적 소견 및 절편내 in situ hybridization 바이러스 동정)

  • Kim, Soon-bok;Song, Geun-suk;Moon, Oun-kyong;Jeong, Chang-geun
    • Korean Journal of Veterinary Research
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    • v.35 no.2
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    • pp.369-374
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    • 1995
  • Aujeszky's disease virus(ADV) was inoculated intranasally into the Korean native goats to investigate pathological findings and pathogenesis of ADV infection by using of histological and immunohistochemical methods and in situ hybridization(ISH). Clinical signs of salvation, pyrexia, pruritus and staggering were followed by death with five days after inoculation, Pathoanatomical findings were edema of the lung and the urinary bladder with hemorrhage and congestion, petechial hemorrhages on the endo-and epicardium, renal congestion, moderate splenomegaly and cystic edema. Main microsocpic lesions observed in all infected goats were confined to the CNS and charcterized by perivascular cuffing with lymphocytes and macrophages, focal gliosis, neuronal degeneration and necrosis, and intranuclear inclusion bodies in the neurons and glial cells. Positive reactions to ADV were detected more frequently in the nuclei than in the cytoplasms of infected nerve cells in the CNS by immunohistochemistry and ISH. Frequenctly localized sites of ADV in the CNS were olfactory bulb, prietal cortex, callosal sulcus and corpus callosum. Positive reactions were also detected in the tonsillar epithelium, and alveolar macrophage and sloughed epithelium of the lung.

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Fracture behavior of monotype and hybrid fiber reinforced self-compacting concrete at different temperatures

  • Mazloom, Moosa;Karimpanah, Hemin;Karamloo, Mohammad
    • Advances in concrete construction
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    • v.9 no.4
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    • pp.375-386
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    • 2020
  • In the present study, the effect of basalt, glass, and hybrid glass-basalt fibers on mechanical properties and fracture behavior of self-compacting concrete (SCC) mixes have been assessed at room and elevated temperatures. To do so, twelve mix compositions have been prepared such that the proper workability, flowability, and passing ability have been achieved. Besides, to make comparison possible, water to binder ratio and the amount of solid contents were kept constant. Four fiber dosages of 0.5, 1, 1.5, and 2% (by concrete volume) were considered for monotype fiber reinforced mixes, while the total amount of fiber were kept 1% for hybrid fiber reinforced mixes. Three different portions of glass and basalt fiber were considered for hybridization of fibers to show the best cocktail for hybrid basalt-glass fiber. Test results indicated that the fracture energy of mix is highly dependent on both fiber dosage and temperature. Moreover, the hybrid fiber reinforced mixes showed the highest fracture energies in comparison with monotype fiber reinforced specimens with 1% fiber volume fraction. In general, hybridization has played a leading role in the improvement of mechanical properties and fracture behavior of mixes, while compared to monotype fiber reinforced specimens, hybridization has led to lower amounts of compressive strength.

Detection of Cytomegalovirus in Atherosclerotic Aorta and Coronary Artery by In Situ Hybridization and PCR

  • Eom, Yong-Bin;Kwon, Tae-Jung;Lee, Sang-Yong;Lee, Won-Tae;Kim, Jong-Bae
    • Biomedical Science Letters
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    • v.8 no.4
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    • pp.257-268
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    • 2002
  • Chronic infection and inflammation have recently been implicated as important etiologic agents of atherosclerosis. Several agents have been suggested as possible candidates including cytomegalovirus (CMV), herpes simplex vims type 1 (HSV-1), Epstein-Barr virus (EBV), Chlamydia pneumoniae, and Helicobacter pylori. We evaluated the relationship between cytornegalovirus infection and atherosclerosis by in situ hybridization and polymerase chain reaction (PCR). We examined 23 subjects with atherosclerosis and 10 matched control subjects without atherosclerosis. CMV was detected by in situ hybridization in 60.9% (14/23) of aorta and 42.9% (9/21) of coronary arteries in subjects with atherosclerosis. It was also detected by PCR in 65.2% (15/23) of aorta and 52.4% (11/21) of coronary arteries. CMV was detected on areas showing early or advanced atheromatous changes. Cells morphologically identical to smooth muscle cells, endothelial cells, lymphocytes, fibroblasts, and Schwann cells were positively reacted with the CMV probe. However. none of the cells to which the probe hybridized contained inclusion bodies, thus strongly suggesting that the arterial wall may be a site of CMV latency. This result Indicates that CMV may potentially play a direct or indirect role in the pathogenesis of human atherosclerosis.

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Detection of Avian Influenza-DNA Hybridization Using Wavelength-scanning Surface Plasmon Resonance Biosensor

  • Kim, Shin-Ae;Kim, Sung-June;Lee, Sang-Hun;Park, Tai-Hyun;Byun, Kyung-Min;Kim, Sung-Guk;Shuler, Michael L.
    • Journal of the Optical Society of Korea
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    • v.13 no.3
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    • pp.392-397
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    • 2009
  • We designed a wavelength interrogation-based surface plasmon resonance (SPR) biosensor to detect avian influenza DNA (AI-DNA). Hybridization reactions between target AI-DNA probes and capture probes immobilized on a gold surface were monitored quantitatively by measuring the resonance wavelength in the visible waveband. The experimental results were consistent with numerical calculations. Although the SPR detection technique does not require the DNA to be labeled, we also evaluated fluorescently-labeled targets to verify the hybridization behavior of the AI-DNA. Changes in resonance were found to be linearly proportional to the amount of bound analyte. A wavelength interrogation-type SPR biosensor can be used for rapid measurement and high-throughput detection of highly pathogenic AI viruses.

Rapid and Easy Diagnosis of Rabbit Haemorrhagic Disease by In Situ Hybridization (In situ Hybridization에 의한 토끼출혈증(rabbit haemorrhagic disease)의 신속.간편한 진단)

  • Park, Nam-Yong;Cho, Ho-Seong;Cho, Kyoung-Oh;Kim, Sang-Jip;Park, Hyung-Seon
    • Korean Journal of Veterinary Pathology
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    • v.5 no.2
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    • pp.57-62
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    • 2001
  • Recently various molecular diagnostic techniques have been used to identify rabbit hemorrhagic disease virus (RHDV), a causative agent responsible for acute hepatitis and disseminated intravascular coagulation in rabbit. But they were hard to perform and time consuming. To detect RHDV in a rapid and easy way, we developed biotinylated oligonucleotide probe within ORF 1 region encoding the polyprotein of RHDV in formalin-fixed and paraffin-embedded tissues from various tissues of 20 rabbits naturally infected with RHDV, Our in situ hybridization (ISH) was quickly carried out within two hours by MicroProbe capillary action system. The ISH produced a positive reaction in liver, kidney and lung. In conclusion, ISH with a biotintlated oligonucleotide probe provided a useful diagnostic method for detecting RHDV.

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Localization of Transferrin mRNA in Rat by DNA/RNA Hybridization (DNA/RNA Hybridization에 의한 흰쥐의 Transferrin mRNA 분포에 관한 연구)

  • Kim, Se-Eun;Kim, Sun-Yeou;Park, Mi-Jung;Song, Jin-Ho;Lee, Eun-Bang;Lee, Heun-Pa;Kim, Young-Choong
    • YAKHAK HOEJI
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    • v.33 no.5
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    • pp.300-307
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    • 1989
  • Expression of transferrin gene in various organs of rat was studied using rat transferrin cDNA. The hybridization method of $[^{35}S]-labeled$ transferrin cDNA with transferrin mRNA in cytoplasmic preparations was used to measure the level of transferrin mRNA. The rat from 15-day old fetus to 21-day old postnatal were employed as an animal model. In the liver, the level of transferrin mRNA increased with increasing age. However, the level of transferrin mRNA in brain was significantly lower than that in liver and the level did not increase with age.

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Flanking Sequence and Copy-Number Analysis of Transformation Events by Integrating Next-Generation Sequencing Technology with Southern Blot Hybridization

  • Qin, Yang;Woo, Hee-Jong;Shin, Kong-Sik;Lim, Myung-Ho;Cho, Hyun-Suk;Lee, Seong-Kon
    • Plant Breeding and Biotechnology
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    • v.5 no.4
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    • pp.269-281
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    • 2017
  • With the continual development of genetically modified (GM) crops, it has become necessary to develop detailed and effective molecular characterization methods to select candidate events from a large pool of transformation events. Relative to traditional molecular analysis methods such as the polymerase chain reaction (PCR) and Southern blot hybridization, next generation sequencing (NGS) technology for whole-genome sequencing of complex crop genomes had proven comparatively useful for in-depth molecular characterization. In this study, four transformation events, including one in Bacillus thuringiensis (Bt)-resistant rice, one in resveratrol-producing rice, and two in beta-carotene-enhanced soybeans, were selected for molecular characterization. To merge NGS analysis and Southern blot-hybridization results, we confirmed the transgene insertion sites, insertion construction, and insertion numbers of these four transformation events. In addition, the read-coverage depth assessed by NGS analysis for inserted genes might provide consistent results in terms of inserted T-DNA numbers in case of complex insertion structures and highly duplicated donor genomes; however, PCR-based methods can produce incorrect conclusions. Our combined method provides an effective and complete analytical approach for whole-genome visual inspection of transformation events that require biosafety assessment.

Influence of Stacking Sequence on Carbon Fiber/Aramid Fiber Hybrid Composite (탄소섬유/아라미드섬유 하이브리드 복합재료의 적층 순서의 영향 평가)

  • Hyeonho Lee;Seoyeon Bae;Sungbi Lee;Myoung-Gyu Lee;Wonjin Na
    • Composites Research
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    • v.36 no.6
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    • pp.383-387
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    • 2023
  • Carbon fiber-reinforced composites have excellent mechanical properties. However, the fracture toughness is a disadvantage due to brittle failure mode. The fracture toughness can be enhanced using hybridization with large-elongation fibers. In this study, polyamide (aramid) fibers are hybridized with carbon fiber with various stacking sequences. As a result, the Izod impact strength was enhanced by 63% with 25% aramid fiber hybridization. It is also shown that there is an optimal point in laminated composite hybridization, [CF/CAF2/CF]s stacking sequence.