• Microbiology and Biotechnology Letters
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• v.35 no.4
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• pp.272-277
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• 2007

The physiological responses of microorganisms to specific nutrient limitation can be regulated at the transcriptional levels. In this study, in order to develop the Pichia pastoris-derived promoter inducible by nutrient-limited condition, we constructed cDNA libraries using RT-PCR of total RNA from P. pastoris in steady-states of phosphate-limited chemostat with different dilution rates. Various genes were detected from cDNA library. Among these genes, the gene encoding putative sodium/phosphate ($Na^+$/Pi) symporter (NPS), high affinity transporter of phosphate, was detected. It was observed that expression of NPS increased in a manner specific to phosphate-limited condition through Northern blot. Therefore, it is thought that the promoter from NPS gene may have the potential as auto-inducible promoter by phosphate-limited culture condition without inducer.

• Journal of Korean Society of Forest Science
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• v.98 no.6
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• pp.757-763
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• 2009

ISSR PCR technique was applied to investigate genetic relationship among 5 Mountain cultivated ginseng populations (Jinan, Hongcheon, Punggi, Andong and Yeongju) and cDNA libraries of wild ginseng roots were constructed and analyzed functional genes related to morphogenesis via EST. Twenty four ISSR markers tested produced 127 polymorphic loci from 5 regional Mountain cultivated ginseng. Among the regional samples, Yeongju was made 18 polymorphic loci that were the highest level of variations among the cultivated regions. The range of similarity coefficient was 0.46~0.58 and the regional samples of Punggi and Hongcheon, Jinan and Andong were classified to similar groups respectively, whereas Yeongju was shown to be separate group with high level of genetic variation in UPGMA cluster analysis. As a result, there was no relationship according to geographical distance and genetic similarity. Eleven cDNA clones were consisted of 9 known genes and 2 unknown genes analyzed by BLAST program of NCBI. To recognize expression pattern of Homeodomain transcription factor related genes, Northern Blot analysis was performed for wild ginseng's leaf and root. As a result, the gene was only expressed by Mountain wild ginseng root.

• The Journal of Korean Medicine
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• v.18 no.2
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• pp.167-186
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• 1997

The inhibitory activity of Aquillariae Lignum (Thymelaeaceae) on type Ⅰ immediate hypersensitivity of the anaphylactic type in the wistar rat model of passive cutaneous anaphylaxis, an IgE-mediated, mast cell-dependent reaction. Administered orally at 250, 500 mg/kg body weight 1 h before the challenge, Aquillariae Lignum potently inhibited PCA in rats which disodium cromoglycate showed poor inhibitory activity. Aquillariae Lignum inhibited compound 48/80-induced anaphylaxis 100% with a dose of 0.5 g/kg body weight at 1 h before or 5 and 10 min after injection of compound 48/80. Aquillariae Lignum (0.05-1.6 mg/ml) also exhibited the dose-related inhibitory effect on compound 48/80-induced histamine release from rat_peritoneal mast cells. Moreover, it was clearly demonstrated that Aquillariae Lignum and disodium cromoglycate disodium cromoglycate potently inhibited such type Ⅰ allergic reactions as anaphylactic shocks, suggesting that these drugs, at least in part, share the same mechanism of action It is suggested that Aquillariae Lignum may exert a stronger inhibition on the mast cell degranulation process. Since Aquillariae Lignum (1.0 mg/ml) inhibited about 90% of histidine decarboxylase activity, the inhibitory activity of Aquillariae Lignum for histamine release was considered to be derived from the inhibition of histidine decarboxylase activity. It results from increased expression of the mRNA coding for histidine decarboxylase, as assessed by Northern blot analysis after a 12 h incubation to P-815 cells with dexamethasone plus 12-O-tetradecanoylphorbol-13-acetate. The addition of Aquillariae Lignum to P-815 cells with dexamethasone plus 12-O-tetradecanoyl-phorbol-13-acetate, significantly inhibited the histidine decarboxylase gene expression. Tumor necrosis $factor-{\alpha}$ was not constitutively expressed in P-815 cells. Substance P selectively activates the tumor necrosis $factor-{\alpha}$ gene expression in P-815 cells. Aquillariae Lignurm inhibited substance P-induced tumor necrosis $factor-{\alpha}$ gene expression. Furthennore, The effect of Aquillariae Lignum on the mRNA expression of novel protein kinase C ${\delta}$ a major isoform of mast cells, was examined by Northern blot analysis. The expression of novel protein kinase C ${\delta}$ mRNA in the presence of Aquillariae Lignum was significantly lower than in the absence of Aquillariae Lignum. These results suggest the possibility that the inhibition of allergic reaction by Aquillanae Lignum should be regulated by tumor necrosis $factor-{\alpha}$ and novel protein kinase C ${\delta}$.

• Journal of Periodontal and Implant Science
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• v.30 no.2
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• pp.347-360
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• 2000

Bone remodeling results from the combined process of bone resorption and new bone formation which is regulated in part by some of the polypeptide growth factors such as platelet derived growth factor(PDGF), which has been known to be an important local regulator of bone cell activity and participate in normal bone remodeling. This process includes strictly regulated gene expression of several bone matrix proteins such as type I collagen and osteopontin, a 44 kDa phosphorylated glycoprotein, which has important roles in bone formation. The purpose of this study is to evaluate the effecs of PDGF-BB on the mRNA expression of bone matrix protein, type I collagen and osteopontin, in MC3T3- E1 cell culture. Cells were seeded at $5{\times}10^5$ cells in 10 ml of minimum essential medium alpha(${\alpha}-MEM$) containig 10% fetal bovine serum, 10 mM beta glycerophosphate. 0.1, 1, 10 ng/ml PDGF-BB were added to the cells for the day 3, 7, 14, 21, 28 and cultured for 24 hours. Type I collagen cDNA, Hf677, and osteopontin cDNA were used as probes for northern blot analysis. Total cellular RNA was purified at indicated day and northern blot analysis was performed. The results were as follows : Type I collagen mRNA expressions were higher at the day 3 and 7, and lower in the day 14, 21 in the control groups. In the experimental groups, mRNA expressions were increased when 0.1 ng/ml PDGF-BB were added on the day 3, 7, 21, and decreased in dose-dependent manner on the day 14, decreased at all added dose on the day 28. Osteopontin mRNA expressions were highest in the day 21 groups and lowest in the day 14 groups in the control groups. Interesting results were shown in the day 14 and 21 groups. We found that osteopontin mRNA level was increased in dose dependent manner in the day 14 groups, and decreased dose dependent manner in the day 21 groups. In conclusion, PDGF-BB may have various control effects on type I mRNA expression in the growth and differentiation process of MC3T3-E1 cells and may have contrary regulatory effects on osteopontin mRNA expression. For examples, when the baseline level of osteopontin mRNA was low, as in the day 14, PDGF-BB up-regulated osteopontin mRNA expression in dose dependent manner, and when the baseline level was high as in the day 21, PDGF-BB down-regulated dose dependent manner. Thus, it may be useful for clinical application in periodontal regeneration procedure if further study were performed.

• Journal of Animal Science and Technology
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• v.44 no.6
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• pp.677-684
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• 2002

We have investigated the expression patterns of candidates for growth stage specifically expressed genes. The expression patterns of the EPV20, aldolase A, Translationally Controlled Tumor Protein (TCTP) and Adipocyte Differentiation Related Protein (ADRP) were examined by semiquantitative RT-PCR and northern blot analysis in skeletal muscle tissues of Hanwoo, especially in the longissimus dorsi at various growth stages. The EPV20 mRNA was expressed in longissimus dorsi tissue of Hanwoo, but there was no difference of expression levels during growth stages. Though the aldolase A gene was reported to be muscle-specific and regulated at developmental stages, the expression levels of aldolase A mRNA in the longissimus dorsi tissues showed little differences at various growth stages. The expression levels of TCTP which was reported as growth-related protein regulated at translation step were gradually increased during growth of Hanwoo. The expression levels of ADRP mRNA were rapidly increased at 24-month-old longissimus dorsi tissue of Hanwoo, and decreased at 30-month-old. Our data suggest that the ADRP gene show as growth-stage dependent expression and is related to fat deposition within muscular tissue.

• Journal of Plant Biotechnology
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• v.38 no.1
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• pp.42-48
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• 2011

Chloroplast genetic engineering of higher plants offers several unique advantages compared with nuclear genome transformation, such as high levels of transgene expression, a lack of position effect due to site-specific transgene integration by homologous recombination, multigene engineering in a single transformation event and reducing risks of gene flow via pollen due to maternal inheritance. We established a reproducible chloroplast transformation system of potato using a tobacco specific plastid transformation vector, pCtVG (trnI-Prrn-aadA-mgfp-TpsbA-trnA). Stable transgene integration into chloroplast genomes and the homoplasmic state of the transgenome were confirmed by PCR and Southern blot analyses. Northern, immunoblot analysis, and GFP fluorescence imaging revealed high expression and accumulation of GFP in the plastids of potato leaves. This system would provide new opportunities for genetic improvement and mass production of value added foreign proteins in this crop.

• Radiation Oncology Journal
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• v.17 no.3
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• pp.223-229
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• 1999

Purpose : The expression pattern of c-jun by ionizing radiation according to cell growth state (exponential growth vs. stationary phase) and its relationship with cell cycle redistribution were investigated. Materials and Methods : The exponential growth phase (day 4) and stationary phase (day 9) cells were determined from cell growth curve according to the elapse of days in CaSki. The cells were irradiated using 6 MV X-ray with a dose of 2 Gy at a fixed dose rate of 3 Gy/min. Northern blot analysis was peformed with total cellular RNA and cell cycle distribution was analyzed using flow cytometry according to time-course after irradiation. Results : The maximum expression of c-jun occurred 1 hour after irradiation in both exponential growth and stationary phase cells. After then c-jun expression was elevated upto 6 hours in exponential growth phase cells, but the level decreased in stationary phase cells. Movements of cells from G0-G1 to S, G2-M phase after irradiation were higher in exponential growth phase than stationary phase. Conclusion : c-jun may be involved in the regulation of cellular proliferation according to the growth states after irradiation.

• Journal of Life Science
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• v.17 no.2 s.82
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• pp.230-234
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• 2007

Oxygen is required for many important aerobic cellular reactions, it may undergo electrontransfer reactions, which generate highly reactive membrane-toxic intermediates (reactive oxygen species, ROS), such as hydrogen peroxide, singlet oxygen, superoxide radical, hydroxyl radical, hydroperoxyl radical, hydroxy ion. Various mechanisms are available to protect cells against damage caused by oxidative free radicals, including scavenging enzyme systems such as superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx). This antioxidant defense system is a very complex and finely tuned system consisting of enzymes capable of detoxifying oxygen radicals as well as low molecular weight antioxidants. In addition, repair and turnover processes help to minimize subcellular damage resulting from free radical attack. $H_2O_2$,one of the major ROS, is produced at a high rate as a product of normal aerobic metabolism. The primary cellular enzymatic defense systems against $H_2O_2$ are the glutathione redox cycle and catalase. From Northern blot analysis of total RNAs from cultured cell with $H_2O_2$ treatment, various results were obtained. Expression of Cu/Zn SOD decreased when cell passage increased, but the level of the Cu/Zn SOD was scarcely expressed in 35 passage.

• KSBB Journal
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• v.17 no.4
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• pp.370-375
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• 2002

Isolation of a gene and determination of its expression pattern are essential in understanding its function. Among the genes localized in 12ql3, stSG3435 EST was chosen to study its expression pattern. The full-length CDNA was cloned by screening of human brain CDNA library and its sequence was determined by serial deletion followed by automated sequencing of the clones with overlapping fragments. The sequence analysis revealed that stSG 3435 CDNA displayed 100% identity to human MYGI and 86% identity to mouse melanocyte proliferation gene-1 (Gamm 1) originally identified from melanocyte, suggesting that MYGI determined by Northern blot analysis revealed the strongest expression in testes with ubiquitous expression in all the tissues tested. In order to investigate the cellular localization of its protein product, the green fluorescence protein gene was fused into the full-length coding sequence of MYGI, Transfection of the fusion construct followed by confocal microscopy resulted in the green fluorescence signal as a punctate state in cytoplasm indication that MYGI was localized in one of the cellular organelles.

• Journal of Periodontal and Implant Science
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• v.36 no.3
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• pp.613-625
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• 2006

Periodontal ligament(PDL) cells and human gingival fibroblasts(HGFs) play important roles in development, regeneration, normal function, and pathologic alteration. PDL cells and HGFs have the similarity related with general characteristics of fibroblast such as spindle shaped morphology, the presence of vimentin intermediate filament and the synthesis of interstitial collagens and fibronectin. There were many studies about the differences between PDL cells and HGFs, but they were not about whole gene level. In this study, we tried to explain the differences of gene expression profiles between PDL cells and HGFs, and the differences among three individuals by screening gene expression patterns of PDL cells and HGFs, using cDNA microarray. Although there were some variants among three experiments, a set of genes were consistentely and differentially expressed in one cell type. Among 3,063 genes, 49 genes were more highly expressed in PDL cells and 12 genes were more highly expressed in HGFs. The genes related with cell structure and motility were expressed more highly in PDL cells. These are cofilin 1, proteoglycan 1 secretory granule, collagen type I(${\alpha}$ 1), adducin gamma subunit, collagen type III(${\alpha}$ 1), fibronectin, lumican(keratan sulfate proteoglycan), and ${\alpha}$ -smooth muscle actin. Tissue inhibitor of metalloproteinase known as the enzyme controlling extracellular matrix with matrix metalloproteinase is more highly expressed in PDL cells, osteoprotegerin known as osteoclastogenesis inhibitory factor is more highly expressed in HGFs. We performed northern blot to verify cDNA microarray results on selected genes such as tissue inhibitor of metalloproteinase, fibronectin, osteoprogeterin. The result of northern blot analysis showed that each cell expressed the genes in similar pattern with cDNA microarray result. This result indicates that cDNA microarray is a reliable method in screening of gene expression profiles.