• 한국식물병리학회지
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• 제14권6호
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• pp.612-613
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• 1998

The nucleotide sequence of hop stunt viroid HHSVd) Kh strain was sequenced by the reverse transcription and polymerase chain reaction. It consists of 296 nucleotides, and differs by one nucleotide deletion of cytosine at the position of 295 from the HSVd-K strain which consists of 297 nucletoides.

• Fisheries and Aquatic Sciences
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• 제8권3호
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• pp.185-187
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• 2005

We tested the in vivo ability of a DNA chip to detect virus-specific genes from virus-infected olive flounder Paralichthys olivaceus and rainbow trout Oncorhynchus mykiss. Target cDNA was obtained from total RNA of virus infected cell lines by reverse transcription (RT) and was labeled with fluorescent dye (Cy5-dUTP). The results show the successful detection of infectious hematopoietic necrosis virus (IHNV) and viral hemorrhagic septicaemia virus (VHSV) genes in the virus-infected fishes.

• Journal of Korean Medical Science
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• 제33권50호
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• pp.319.1-319.5
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• 2018

The incidence of severe fever with thrombocytopenia syndrome (SFTS) has increased in Korea since a first report in 2013. We investigated whether SFTS existed before 2013 using real-time reverse transcription polymerase chain reaction and stored blood samples from febrile patients with thrombocytopenia. Four cases of SFTS were identified, with the earliest occurring in 2008.

• The Plant Pathology Journal
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• 제39권1호
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• pp.158-158
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• 2023

• 생명과학회지
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• 제27권3호
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• pp.318-324
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• 2017

본 연구는 참깨 70% 에탄올 추출물의 항산화 효과와 미백효과를 검증하여 나타내었다. 참깨 추출물의 전자공여능 측정실험은 $1,000{\mu}g/ml$에서 71.7%의 효과를 나타냈으며, tyrosinase 저해활성 측정은 $1,000{\mu}g/ml$의 농도에서 42%의 효과를 보였다. 참깨 추출물의 세포 생존율을 melanoma cell (B16F10)에서 확인하기 위하여 MTT assay를 진행하였으며, 세포 생존율을 측정한 결과, $1,000{\mu}g/ml$ 농도에서 84.3%의 독성을 보였다. 이하의 세포실험에서는 세포 생존율이 90% 이상되는 농도인 $500{\mu}g/ml$ 이하에서 실험을 진행하였다. 참깨 추출물의 MITF, TRP-1, TRP-2 및 tyrosinase의 단백질 발현 효과를 50, 250, $500{\mu}g/ml$ 농도에서 측정하였으며, 그 결과 MITF, TRP-1, TRP-2 및 tyrosinase의 각각 $500{\mu}g/ml$ 농도에서 68.3%, 39.2%, 89.7%, 22.3%의 단백질 발현 억제 효과를 보였다. 또한, MITF, TRP-1, TRP-2 및 tyrosinase의 mRNA발현을 RT-PCR로 50, 250, $500{\mu}g/ml$ 농도에서 측정하였고, 양성 대조군으로 GAPDH를 사용하였다. 그 결과, 참깨 추출물의 $500{\mu}g/ml$ 농도에서 각각 81.8%, 66.5%, 84.2%, 68.1%의 mRNA 발현이 감소함을 확인할 수 있었다. 이상의 결과로 부터 참깨 추출물의 화장품 소재로서의 가능성을 확인할 수 있었다.

• Animal cells and systems
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• 제3권3호
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• pp.311-317
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• 1999

Prolactin (PRL) was reported to be locally synthesized in many brain areas including the hypothalamus, thalamus (TH) and hippocampus (HIP). In the pituitary lactotrophs, PRL synthesis is dependent upon a pituitary-specific transcription factor, Pit-1. In the present study, we attempted to identify Pit-1 or Pit-1-like protein in brain areas known as the synthetic sites of PRL. Reverse transcription-polymerase chain reaction (RT-PCR) and Northern blot analysis showed the same Pit-1 transcripts in brain areas such as the medial basal hypothalamus (MBH), preoptic area (POA), TH, and HIP with the Pit-1 transcripts in the anterior pituitary (AP). Electrophoretic mobility shift assay (EMSA) was run with nuclear protein extracts from brain tissues using a double strand oligomer probe containing a putative Pit-1 binding domain. Shifted bands were found in EMSA results with nuclear proteins from MBH, POA, TH and HIP. Specific binding of the Pit-1-like protein was further confirmed by competition with an unlabeled cold probe. Antisense Pit-1 oligodeoxynucleotide (Pit-1 ODN), which was designed to bind to the Pit-1 translation initiation site and block Pit-1 biosynthesis, was used to test Pit-1 dependent brain PRL transcription. Two nmol of Pit-1 ODN was introduced into the lateral ventricle of a 60-day old male rat brain. RNA blot hybridization and in situ hybridization indicated a decrease of PRL mRNA signals by the treatment of Pit-1 ODN. Taken together, the present study suggests that Pit-1 may play an important role in the transcriptional regulation of local PRL synthesis in the brain.

• Molecules and Cells
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• 제37권10호
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• pp.734-741
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• 2014

Histone modifications on major transcription factor target genes are one of the major regulatory mechanisms controlling adipogenesis. Plant homeodomain finger 2 (PHF2) is a Jumonji domain-containing protein and is known to demethylate the histone H3K9, a repressive gene marker. To better understand the function of PHF2 in adipocyte differentiation, we constructed stable PHF2 knock-down cells by using the mouse pre-adipocyte cell line 3T3-L1. When induced with adipogenic media, PHF2 knock-down cells showed reduced lipid accumulation compared to control cells. Differential expression using a cDNA microarray revealed significant reduction of metabolic pathway genes in the PHF2 knock-down cell line after differentiation. The reduced expression of major transcription factors and adipokines was confirmed with reverse transcription- quantitative polymerase chain reaction and Western blotting. We further performed co-immunoprecipitation analysis of PHF2 with four major adipogenic transcription factors, and we found that CCATT/enhancer binding protein (C/EBP)${\alpha}$ and C/EBP${\delta}$ physically interact with PHF2. In addition, PHF2 binding to target gene promoters was confirmed with a chromatin immunoprecipitation experiment. Finally, histone H3K9 methylation markers on the PHF2-binding sequences were increased in PHF2 knock-down cells after differentiation. Together, these results demonstrate that PHF2 histone demethylase controls adipogenic gene expression during differentiation.

• IMMUNE NETWORK
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• 제11권4호
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• pp.203-209
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• 2011

Background: T cell immunoglobulin mucin containing molecule (TIM)-3 is expressed in differentiated Th1 cells and is involved in the suppression of the cytokine production by these cells. However, the regulation of the expression of other T cell genes by TIM-3 is unclear. Herein, we attempted to identify differentially expressed genes in cells abundantly expressing TIM-3 compared to cells with low expression of TIM-3. Methods: TIM-3 overexpressing cell clones were established by transfection of Jurkat T cells with TIM-3 expression vector. For screening of differentially expressed genes, gene fishing technology based on reverse transcription-polymerase chain reaction (RT-PCR) using an annealing control primer system was used. The selected candidate genes were validated by semi quantitative and real-time RT-PCR. Results: The transcription of TIMP-1, IFITM1, PAR3 and CCL1 was different between TIM-3 overexpressing cells and control cells. However, only CCL1 transcription was significantly different in cells transiently transfected with TIM3 expression vector compared with control cells. CCL1 transcription was increased in primary human $CD4^+$ T cells abundantly expressing TIM-3 but not in cells with low expression of TIM-3. Conclusion: CCL1 was identified as a differentially transcribed gene in TIM-3-expressing $CD4^+$ T cells.

• Mycobiology
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• 제45권4호
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• pp.362-369
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• 2017

We assessed the regulation of cryparin, a class II hydrophobin, using three representative mitogen-activated protein kinase (MAPK) pathways in Cryphonectria parasitica. Mutation of the CpSlt2 gene, an ortholog of yeast SLT2 in the cell wall integrity (CWI) pathway, resulted in a dramatic decrease in cryparin production. Similarly, a mutant of the CpBck1 gene, a MAP kinase kinase kinase gene in the CWI pathway, showed decreased cryparin production. Additionally, mutation of the cpmk1 gene, an ortholog of yeast HOG1, showed decreased cryparin production. However, mutation of the cpmk2 gene, an ortholog of yeast Kss1/Fus3, showed increased cryparin production. The easy-wet phenotype and accumulation of the cryparin transcript in corresponding mutants were consistent with the cryparin production results. In silico analysis of the promoter region of the cryparin gene revealed the presence of binding motifs related to downstream transcription factors of CWI, HOG1, and pheromone responsive pathways including MADS-box- and Ste12-binding domains. Real-time reverse transcriptase PCR analyses indicated that both CpRlm1, an ortholog of yeast RLM1 in the CWI pathway, and cpst12, an ortholog of yeast STE12 in the mating pathway, showed significantly reduced transcription levels in the mutant strains showing lower cryparin production in C. prasitica. However, the transcription of CpMcm1, an ortholog of yeast MCM1, did not correlate with that of the mutant strains showing downregulation of cryparin. These results indicate that three representative MAPK pathways played a role in regulating cryparin production. However, regulation varied depending on the MAPK pathways: the CWI and HOG1 pathways were stimulatory, whereas the pheromone-responsive MAPK was repressive.

• 생명과학회지
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• 제23권11호
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• pp.1336-1341
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• 2013

아시아에서 한방약초로 널리 알려진 당귀 추출물의 미백활성을 알아보기 위하여 tyrosinase 저해활성을 측정한 결과 1,000 ${\mu}g/ml$의 농도에서 70% 이상의 활성을 나타내었다. 또한 당귀 추출물에 대한 멜라노마 세포(B16F10)의 세포생존율을 확인한 결과 500 ${\mu}g/ml$의 농도에서 99% 이상의 세포생존율을 확인할 수 있었다. 미백 관련 인자인 MITF, TRP-1, TRP-2 및 tyrosinase의 mRNA 발현량을 측정한 결과 50 ${\mu}g/ml$의 농도에서 각각 85.7%, 123.9%, 68.8%, 208%로 당귀 추출물을 처리하지 않은 군보다 감소하였음을 확인할 수 있었다. 이러한 연구 결과에 따라 당귀 추출물이 melanin 합성과 관련이 있는 유전자 발현의 억제효과가 있음을 확인할 수 있었으며, 미백 화장품 소재로서의 가능성을 확인하였다.