• BMB Reports
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• 제41권8호
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• pp.568-574
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• 2008

Antisense RNA molecules are powerful tools for controlling the expression of specific genes but their use in prokaryotes has been limited by their unpredictable antisense effectiveness. Moreover, appreciation of the molecular mechanisms associated with silencing in bacteria is still restricted. Here we report our attempts to define an effective antisense strategy in E. coli, and to dissect the observed silencing process. Antisense constructs complementary to different regions of lacZ were investigated, and silencing was observed exclusively upon expression of antisense RNA hybridising the 5'UTR of lac messenger. The level of lacZ mRNA was reduced upon expression of this antisense construct, and the silencing competence was found to be closely associated with its stability. These observations may help in the design of antisense molecules directed against prokaryotic genes.

• 대한임상검사과학회지
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• 제47권4호
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• pp.298-305
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• 2015

최근 종양을 극복하고자 하는 새로운 접근 방법가운데 하나로, 종양세포내에 발현되는 줄기세포 전사인자들(Oct4, Sox2, KLF4 and Lin28)을 억제하여 종양을 치료하는 연구들이 증가하고 있다. 본 실험은 미분화 전사인자를 표적(조절)하는 microRNA-126을 이용하여 난소종양세포들(6종: HSC832(t)c, Ovcar3, Skov3, PA-1, TOV21G and Tov112D)들 생존과 성장에 어떠한 생물학적 변화를 유도하는지 연구하였다. Scramble과 microRNA-126를 난소종양세포들에 처리 후 세포모양 관찰결과 Skov3를 제외한 난소 종양세포들에서 형태학적 모양 변성과 부유현상을 관찰하였다. CCK-8을 이용한 세포분열능 분석에서 Skov3를 제외한 난소 종양세포들의 분열능력이 점차적으로 감소되는 것을 확인하였다. 특히 Tov112D, Tov21G and PA-1에서 각 시간대별로 뚜렷한 세포분열 능력 감소를 확인할 수 있었다. RT-PCR결과 미분화 전사인자들(Sox2, Lin28)의 발현감소를 확인할 수 있었다. 이러한 결과들은 microRNA-126이 다양한 난소 종양세포들을 표적하여 세포분열능과 사멸을 유도할 수 있는 가역적 환경(유전자 발현조절)을 제공함과 동시에 임상 치료에 대한 분자생물학적 단서를 제공할 수 있을 것이다.

• 한국동물학회지
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• 제28권2호
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• pp.108-119
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• 1985

Stress protein (SP) 遺傳子發現의 調節機構를 밝히기 위한 한가지 방편으로 환경의 stress와 pH가 SP의 合成誘導에 어떤 作用을 하는지를 SDS-PAGE를 이용해서 分析하였다. 蛋白質合成의 전반적 양상은 MEF와 SCK 세포에서 달랐으나 SP의 양상은 동일하였다. 그중에서 $SP_70$의 誘導와 感衰의 kinetics는 특히 흥미로웠다. $SP_70$의 kinetics는 酸性 pH와 正常 pH에서 類似하였으나 最大量의 SP 合成에 필요한 溫度와 그 處理時間은 pH에 의해 달리 나타나서, 酸性 pH 에서는 자은 溫度와 짧은 處理時間에서 나타나고 더욱 오래 지속되는 경향을 보였다. SP의 合成誘導와 SP mRNA의 축적은 actinomycin D에 의해 阻止되는 사실로 미루어 SP의 合成이 誘導되기 위해서는 새로운 mRNA의 合成이 필요함을 알수 있었고, cycloheximide 처리의 결과는 SP의 合成誘導에 앞서서 어떤 特異한 蛋白質의 合成은 일어나지는 않음을 알수 있었다. 이상과 같은 몇가지 實驗結果는 MEF와 SCK 세포에서 SP의 合成誘導는 일차적으로 轉寫水潗에서 調節되며, $SP_70$의 合成은 自動調節됨과 아울러 SP의 水潗은 세포늬 stress 상태와 相關關係가 있는 것으로 推論할수 있음을 보여주었다.

• 생명과학회지
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• 제18권3호
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• pp.374-380
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• 2008

본 연구는 무균성수막염 의심환자의 다양한 검체로부터 enterovirus의 진단을 위하여 real-time NASBA, 2 step RT-PCR 시험과 세포배양 시험을 각각 실시하여 각 시험법의 검출율, 특이도, 사용자 편리성, 시험소요 시간, 교차오염의 가능성 등을 비교 검토하였다. 비교시험 결과 전체 292건의 검체로부터 real-time NASBA에서 145건, 세포배양에서 101건, 2 step RT-PCR에서 86건이 양성으로 나타나 real-time NASBA가 가장 검출율이 높은 시험법임을 알 수 있었다. Enterovirus 외의 무균성수막염 원인바이러스에 대한 특이도 비교 시험결과 2 step RT-PCR 시험에서 rhinovirus 10건 중 1건이 위양성 반응을 나타내어 다른 시험법에 비해 특이도가 떨어지는 것으로 나타났다. Real-time NASBA는 하나의 튜브에서 증폭과 검출이 동시에 일어나 다른 시험과 비교하여 교차오염의 가능성이 낮으며 또한 시험 소요시간이 5시간 정도로 세포배양(5-14일 소요) 및 2 step RT-PCR(9시간소요) 에 비하여 신속하게 진단할 수 있어 일선병원이나 실험실에서 enterovirus를 검출을 위하여 적용할 수 있을 것으로 사료된다.

• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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• pp.79.1-79
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• 2003

Two pathogen-induced hot pepper transcription factors (CaNACl and CapIfl) were introduced into‘MicroTom’tomato by Agrobacterium tumefaciens-mediated transformation. We used to nptII containing kanamycin resistance gene as a selection marker. Both transformed and non-transformed plants were transferred to pot after rooting test in vitro. To approximate the levels of caNACl transcript in leaves of wild-type and transgenic plants, RNA blots were hybridized with double-stranded full-length CaNACl probe at moderate stringency, Although the relative signal strength for hybridization fluctuated among the samples on different blots, transgenic plant lines N-1, N-2 and N-3 consistently displayed increased levels of CaNACl transcript relative to other transgenic lines and wild-type plants. Of all the transgenic lines examined, line N-7 had the least amount of CaNACl transcript. Role of these transcription factors in pathogen defense will be examined by overexpression in tomato.

• Journal of Microbiology
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• 제33권2호
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• pp.149-153
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• 1995

Mitochondrial DNA has been isolated from Trimorphomyces papilionaceus. By analyzing DNA fragments digested by restriction enzymes, a restriction site map has been constructured. The mtDNA of T. papilionaceus amounts to 48.5 kb in size and is circular in structure. Entire mitochondrial DNA was cloned in E coli plasmids and Northern blot hybridization was done using cloned and subcloned DNAs as probes. Based on hybridization results of mitochondrial RNA transcripts, a transcription map was prepared.

• Molecules and Cells
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• 제40권3호
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• pp.169-177
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• 2017

Tissue-specific transcription is critical for normal development, and abnormalities causing undesirable gene expression may lead to diseases such as cancer. Such highly organized transcription is controlled by enhancers with specific DNA sequences recognized by transcription factors. Enhancers are associated with chromatin modifications that are distinct epigenetic features in a tissue-specific manner. Recently, super-enhancers comprising enhancer clusters co-occupied by lineage-specific factors have been identified in diverse cell types such as adipocytes, hair follicle stem cells, and mammary epithelial cells. In addition, noncoding RNAs, named eRNAs, are synthesized at super-enhancer regions before their target genes are transcribed. Many functional studies revealed that super-enhancers and eRNAs are essential for the regulation of tissue-specific gene expression. In this review, we summarize recent findings concerning enhancer function in tissue-specific gene regulation and cancer development.

• Journal of Korean Dental Science
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• 제6권1호
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• pp.22-26
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• 2013

Purpose: Transglutaminase 2 (TGase 2) is expressed by tumor necrosis factor-${\alpha}$ in various carcinoma. The role of TGase 2 expression in salivary gland tumors is not clear yet. Established slaivary gland tumor (SGT)cell line has been used to study the pathogenesis of salivary gland adenocarcinoma on a cellular level in vitro. The pupose of this study were to examine mRNA expression of TGase 2 in SGT cell line compared to other tumor cell lines, and to apply these results to the pathogenesis of salivary gland tumor. Materials and Methods: After SGT, SCC-15, HN 4, and HeLa tumor cell lines were cultured under preconfl uency, and 3 days after postconfl uency, the cells were harvested for total RNA extraction and cDNA preparation. Result: Reverse transcription polymerase chain reaction for semiquantitative mRNA analysis was done. TGase 2 mRNA expression was not induced by confl uency in all the cell lines. TGase 2 mRNA expression was variable but markedly enhanced in SGT cell line. Conclusion: mRNA expression of TGase 2 should play an important role in the pathogenesis of SGT cell line originated from ductal cell.

• Animal cells and systems
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• 제13권3호
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• pp.297-304
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• 2009

A complete mRNA sequence of transferrin from the wax moth, Galleria mellonella, was obtained, and compared with those of other species. We previously reported that the sequence was most similar to those of Manduca sexta and Bombyx mori. As in other moths, G. mellonella transferrin had only one iron-binding site at its N-terminal region. Semi-qRT PCR was conducted to investigate tissue-specific distribution and transcriptional regulation of the wax moth transferrin mRNA. Larval muscle and fat body contained larger quantity of mRNA than other tested tissues. In this study, it was observed that iron and cadmium regulated transferrin transcription, and this regulation pattern was tissue specific. Iron up-regulated transferrin mRNA level in fat body, while suppressed it in the Malpighian tubules and silk glands. Cadmium decreased the mRNA level in fat body, muscle, and Malpighian tubules, but significantly increased the mRNA level in silk glands. In addition, the mRNA expression was induced by all tested pathogen-associated molecular patterns (PAMPs) including LPS, lipoteichoic acid (LTA), glucan, and even chitin.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2001년도 춘계학술발표대회
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• pp.29-29
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• 2001

This study was carried out to examine, by the reverse transcription chain reaction(RT-PCR)and Immunostain assays, epidermal growth factor mRNA expression in bovine ova during oocyte maturation in vitro(0-2lh)and after fertilization in vitro(6-144hr: zygotes to blastocysts). In this study, the transcripts of EGF was detected in oocytes using primers for EGF. Transcripts for EGF mRNA was not detected in oocytes through in vitro maturation. But EGF mRNA were present after fertilization up to the 2-cell stage and the blastocyst stage. The highest mRNA levels in 4-cell stage embryos were decreased at 8cell stage and then reincreased upto morulae and blastocysts. The results of this study showed EGF mRNA are present in embryo after fertilization and this factors are involved in the regulation of bovine embryo development.