• The Plant Pathology Journal
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• 제26권3호
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• pp.286-288
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• 2010

Plant tissue homogenization using a mortar or mechanical equipment has been the preferred method for obtaining high yields of total RNA; this method, however, is both time-consuming and expensive. Additionally, homogenization may generate excessive endogenous RNases, polyphenolics, and other substances that reduce the quality and quantity of RNA. In this study, we describe the microwave irradiation-assisted RNA extraction (MIRE) technique which, without tissue disruption and homogenization, allows for the cost-effective and rapid generation of intact RNA from apple cane shavings and the reliable detection of apple virus by RT-PCR.

• Toxicological Research
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• 제20권1호
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• pp.31-36
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• 2004

Against environmental stress, ceruloplasmin which is a plasma protein, are believed to play central roles in antioxidant- or peroxidase-activity in blood stream to remove free radicals, which may be caused by exposing of $\gamma$-irradiation. In human U-937 cells exposed to $\gamma$-irradiation, the levels of mRNA in ceruloplasmin gene were measured on 0, 4, 12, 24 hr after exposing by using comparative RT-PCR (Reverse transcriptase-polymerase chain reaction) which was achieved to compare with house keeping genes such as $\beta$-actin and hprt. After $\gamma$-irradiation of 100 rads or 200 rads, the total quantities of RNA were increased as dose and time dependent manner. On the contrary, the variation of mRNA expression in ceruloplasmin was not found until 4 hr after irradiation. After 12 hr and 24 hr of irradiation, the levels of mRNA in ceruloplasmin were significantly increased as dose and time dependent manner than un-exposed cells.

• 한국식물학회:학술대회논문집
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• 한국식물학회 1987년도 식물생명공학 심포지움 논문집 Proceedings of Symposia on Plant Biotechnology
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• pp.357-389
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• 1987

In an attempt to clarify the physiological functions of individual isoperoxidases, we have studied enzymatic and immunological properties as well as cellular distribution of isoperoxidases from tobacco callus and Korean radish. The gene expression patterns of isoperoxidases in shoot and non-shoot-forming tobbaco callus were also examined by rabbit reticulocyte lysatein vitro translation system. These results indicate that fraction of translatable poly(A)-isoperoxidase mRNA was increased considerably in shoots. At the present time, at least 6-7 isoperoxidases could be detected from the translation mixture of total cellular RNA, among which only one cell wall localized anodic isoperoxidase (named A3) mRNA was bimorphic mRNA. These data suggest the possible regulation of peroxidase activity during shoot formation by altering the polyadenylation state of mRNA. In case of Korean radish seedlings, poly(A)- peroxidase mRNA were also increased depending upon aging.

• 한국생물정보학회:학술대회논문집
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• 한국생물정보시스템생물학회 2003년도 제2차 연례학술대회 발표논문집
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• pp.139-146
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• 2003

암의 조기 발견 및 예후 예측을 위하여 마이크로어레이 데이터를 이용할 수 있다. 하지만 이를 분석하기 위해서는 40${\mu}g$ 이상의 RNA 샘플이 필요한데, 실제 임상 시료를 사용하는 경우 요구되는 충분한 양을 얻기가 어려운 단점이 있다. 따라서 소량의 RNA 샘플을 채취한 후 PCR 증폭 과정을 통하여 요구되는 양의 샘플을 얻을 수 있는 RNA 증폭 방법이 시도되고 있고, 이를 마이크로어레이 실험에 이용하기 위해서는 증폭 전후의 유사성이 보장되어야 한다. 본 논문에서는 증폭 RNA와 전체 RNA의 유사성을 비교하기 위한 새로운 방법으로 엔트로피 기반의 방법을 제시한다. 아울러 다양한 조건에 따라서 엔트로피값을 측정하여 세포주와 조직에서 엔트로피 값이 어떻게 사용될 수 있는지 체계적인 분석을 하였다.

• 한국어류학회지
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• 제22권4호
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• pp.267-272
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• 2010

문치가자미 Pleuronectes yokohamae의 RNA/DNA비 일주기 변화를 조사하기 위하여 부화후 89일, 전장 $23.2{\pm}0.2mm$의 치어를 48시간 동안 일정한 시간 간격으로 채집하였다. RNA/DNA비는 주간(0800, 1100, 1400, 1700)에 높았고 야간(2000, 2300, 0200, 0500 hours)에는 낮게 나타났다. 이번 연구결과에서 문치가자미의 생화학적 일주기 변화를 확인하였고, 이와 같은 RNA/DNA비의 일주기적 변화를 고려하여 시료 채집시간 결정을 포함한 시료채집 계획작성과 data분석이 필요할 것으로 생각된다.

• Journal of Microbiology and Biotechnology
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• 제5권4호
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• pp.194-199
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• 1995

A portion (7.4 kb) of ribosomal DNA tandem repeat unit from a genome of the mushroom T. matsutake has been cloned. A 1.75 kb EcoRI fragment was cloned first using S. cerevisiae 255 rRNA gene as a probe, and this was then used for further cloning. A chromosomal walking experiment was carried out and the upstream region of the 1.75 kb fragment was cloned using SmaI/BamHI enzyme, the size was estimated to be 5.2 kb in length. Part of the downstream region of the 1.75 kb fragment was also cloned using XbaI/BamHI enzymes. Restriction enzyme maps of three cloned DNA fragments were constructed. Northern hybridization, using total RNA of T. matsutake, and the restriction fragments of three cloned DNAs as probes, revealed that all four ribosomal RNA genes (large subunit[LSU], small subunit [SSU], 5.85 and 5S rRNA genes) are present in the cloned region. The gene organization of the rDNA are regarded as an intergenic spacer [IGS]2 (partial) - SSU rRNA - internal transcribed spacer [ITS]1 - 5.8S rRNA - ITS2 - LSU rRNA - IGS1 -5S rRNA - IG52 (partial).

• 대한환경공학회지
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• 제22권5호
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• pp.939-947
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• 2000

연속 회분식 반응기를 이용하여 생물학적 인 제거에 관한 미생물 분포 연구를 수행하였다. 탄소원으로 초산을 넣은 합성 폐수를 사용하였고 미생물 체류 시간과 수리학적 체류 시간은 각각 10일과 16시간으로 유지하였다. 인 방출과 흡수가 운전 시간이 경과됨에 따라 점점 빠르게 일어났으며 약 200일 경과 후 안정적인 인 제거가 유지되었다. 안정적인 생물학적 인 제거가 유지될 때의 미생물 분포를 조사하기 위하여 17개의 ribosomal RNA (rRNA) signature probe를 합성하여 슬러지로부터 분리한 전체 rRNA에 대하여 slot hybridization을 실시하였다. 분리한 전체 RNA에는 proteobacteria의 베타군 (beta subclass)에 속하는 rRNA가 가장 많이 함유되어 있음을 확인하였고 CTE probe와 관계된 rRNA가 다음으로 많이 분포하였다. 전통적으로 생물학적 인 제거를 담당하는 미생물로 여겨져 왔던 Acinetobacter, Aeromonas, Pseudomonas의 rRNA는 10% 미만으로 존재하고 있음이 확인되었다. 이러한 결과로부터 Rhodocyclus 그룹같은 proteobacteria의 베타군과 CTE에 속하는 미생물이 인 제거에 중요한 역할을 수행할 것으로 생각되었고 Acinetobacter, Aeromonas, Pseudomonas 등은 생물학적 인 제거에 있어서 과평가된 것으로 판단되었다.

• BMB Reports
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• 제36권3호
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• pp.312-318
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• 2003

The purpose of this study was to investigate the effects of altering relative intakes of fat and carbohydrates on serum lipid profiles, hepatic acyl-CoA synthetase (ACS), carnitine palmitoyltransferase-I (CPT-I), and the acetyl-CoA carboxlyase (ACC) mRNA level in Sprague-Dawley rats. For four weeks the rats were fed either an AIN-76 diet or one of its modified diets that were supplemented with 20% beef tallow (high-fat diet, HF) and 66.3% sucrose (highsucrose diet, HS). The HS group had significantly higher serum triglyceride and total cholesterol concentrations when compared with the other groups. Serum LDL-cholesterol concentrations in the HS and HF groups were significantly higher when compared to the normal diet (ND) group. Serum HDL-cholesterol levels of the ND and HS groups were significantly higher than those of the HF group. The hepatic total lipid level of the HF group was significantly higher than those of other groups; triglyceride levels of the HS and HF groups were significantly higher than those of the ND group. Hepatic ACS mRNA levels of the HF group were significantly higher than those of the ND group. Hepatic CPT-I mRNA levels were higher in the HF group than other groups. Also, ACC mRNA levels in the liver increased in the HF group. In conclusion, changes in the composition of dietary fat and carbohydrates could affect the hepatic ACS, CPT-I, and ACC mRNA levels. These results facilitate our understanding of the coordinated regulation of the ACS, CPT-I, and ACC mRNA levels and will serve to enhance our understanding of the molecular mechanisms that underlie the regulation of fatty acid metabolism.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제31권5호
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• pp.363-369
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• 2005

Purpose: To determine the role of Insulin-like Growth Factor-I (IGF-I) in the regulation of Vascular Endothelial Growth Factor (VEGF) expression in MG-63 cells and then to find the mechanism b which this regulation occurs. Materials and methods: MG-63 cells were grown to confluence in 60-mm dishes. To determine the effects of IGF-I on expression of VEGF mRNA according to time and concentration, the cells were treated with 10 nM IGF-I, following isolation of total RNA and Northern blot analysis after 1, 2, 4, 8, 12, 24 hours and after 2 hours of treatment with 0.5, 2, 10, 25, 50 nM IGF-I respectively, isolation of total RNA and Northern blot analysis were followed. To determine the mechanism of action of IGF-I, inhibitors such as hydroxyurea $(76.1\;{\mu}g/ml)$, actinomycin D $(2.5\;{\mu}g/ml)$, cycloheximide $(10\;{\mu}g/ml)$ were added 1 hour after treatment of 10 nM IGF-I. Results: 1. the expression of VEGF mRNA was increased with treatment of IGF-I. 2. The expression of VEGF mRNA was increased according to time-and concentration dependent manner of IGF-I. 3. The effect of IGF-I was decreased by hydroxyuera, actinomycin D, but not by cycloheximide. Conclusion: IGF-I regulate the expression of VEGF mRNA in the level of DNA synthesis and transcription. These results could suggest that IGF-I plays an important role in angiogenesis in the process of new bone formation and remodeling.

• Journal of Plant Biology
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• 제38권2호
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• pp.203-209
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• 1995

담배(Nicotiana tobacum L. cv. Wisconsine 38) 잎 절편을 해녀콩(Canavalia lineata)의 leghemoglobin(Lb) cDNA를 포함하는 Agrobacterium과 함께 배양한 후, 0.5mg/L BAP, 0.1mg/L ${\alpha}-NAA$와 200mg/L kanamycin, 500 mg/L carbenicillin을 포함하는 MS 배지에서 선별하여 7개의 재분화 개체를 얻었다. 이로부터 분리한 게놈 DNA에 대한 Southern 혼성화 반응과 PCR 결과로 Lb cDNA가 담배의 게놈에 삽입되었음을 확인하였다. 형질전환된 담배로부터 분리한 RNA에 대한 northern 혼성환 실험 결과 약 1,000 nt의 RNA가 혼성화 반응을 보였으며, 총 RNA를 주형으로 합성한 1차 가닥의 cDNA를 PCR로 증폭한 결과, Lb cDNA와 혼성화 반응을 보이는 0.5 kb의 DNA가 증폭되었다. 콩의 Lb에 대한 다군항체(polyclonal antibody)를 사용하여 단백질 면역 항체 반응을 실시한 결과, Lb로 판단되는 약 15.8 kD의 위치에서 혼성화 반응이 나타났다. 이상의 결과로 해녀콩 Lb cDNA가 형질전환된 담배의 게놈에 삽입되었을 뿐만 아니라 mRNA로 전사되어 Lb 단백질로 해독되었음을 알 수 있었다.