• Journal of Plant Biotechnology
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• v.38 no.1
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• pp.62-68
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• 2011

Calmodulin (CaM), a $Ca^{2+}$ binding protein in eukaryotes, mediates cellular $Ca^{2+}$ signals in response to a variety of biotic and abiotic external stimuli. The $Ca^{2+}$-bound CaM transduces signals by modulating the activities of numerous CaM-binding proteins. As a CaM binding protein, AtCBP63 ($\b{A}$rabidopsis thaliana $\b{C}$aM-binding protein $\underline{63}$ kD) has been known to be positively involved in plant defense signaling pathway. To investigate the pathogen resistance function of AtCBP63 in potato, we constructed transgenic potato (Solanum tuberosum L.) plants constitutively overexpressing AtCBP63 under the control of cauliflower mosaic virus (CaMV) 35S promoter. The overexpression of the AtCBP63 in potato plants resulted in the high level induction of pathogenesis-related (PR) genes such as PR-2, PR-3 and PR-5. In addition, the AtCBP63 transgenic potato showed significantly enhanced resistance against a pathogen causing bacterial soft rot, Erwinia carotovora ssp. Carotovora (ECC). These results suggest that a CaM binding protein from Arabidopsis, AtCBP63, plays a positive role in pathogen resistance in potato.

• Archives of Plastic Surgery
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• v.33 no.1
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• pp.39-45
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• 2006

This study is to examine the relationship between TGF-b1 expression and CTGF expression, and to evaluate the effect of Sp1 blockade on the expression of TGF-b1, CTGF and extracellular genes, clones of fibroblasts stably transfected with Sp1 decoy ODN. R-Sp1 decoy ODN was highly resistant to degradation by nucleases or serum, compared to the linear or phosphorothioated-Sp1 decoy ODN. Skin wounds were created on the back of 36 anesthetized rats. They were divided into four groups-the rats with normal skin, with wounded skin without decoy, with wounded skin injected with R-Sp1 decoy, and with wounded skin injected with mismatched R-Sp1 decoy, respectively. Skins were collected at 3rd, 5th, 7th, 14th day after wounding. Cellular RNA was extracted by RT-PCR analysis. TGF-${\beta}1$ and CTGF were deeply related with skin fibrosis during scar formation and it appeared that TGF-${\beta}1$ may cause the induction of CTGF expression. R-Sp1 decoy ODN inhibited TGF-${\beta}1$ and CTGF expression both in cultured fibroblasts and in the skin of rats. These results indicate that targeting Sp1 with R-type decoy efficiently blocks extracellular matrix gene expression, and suggest an important new therapeutic approach to control the scarring in normal wound healing and fibrotic disorders.

• Korean Journal of Veterinary Research
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• v.44 no.2
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• pp.207-215
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• 2004

It is well known that the hypothalamic-pituitary-adrenocortical (HPA) axis is under the negative feedback control of adrenal corticosteroids. Previous studies have suggested that glucocorticoids can regulate neuroendocrine cells in the paraventricular nucleus (PVN) by modulating catecholaminergic transmission, a major excitatory modulator of the HPA axis at the hypothalamic level. But, the effects of corticosteroids on the expression of adrenoceptor subtypes are not fully understood. In this work, we examined mRNA levels of six adrenoceptor subtypes (${\alpha}_{1A}$, ${\alpha}_{1B}$, ${\alpha}_{2A}$, ${\alpha}_{2B}$, ${\beta}_1$ and ${\beta}_2$) in the PVN of normal and adrenalectomized (ADX) rats. Total RNA ($2.5{\mu}g$) was extracted from PVN micropunches of brain slices ($500{\mu}m$) and analyzed by reverse transcription-polymerase chain reaction (RT-PCR). The levels of corticotropin-releasing hormone (CRH) mRNA were increased in the ADX rats relative to normal rats, indicating that the PVN had been liberated from the negative feedback of corticosteroids. Among the six adrenoceptor subtypes examined, mRNA levels for ${\alpha}_{1B}$- and ${\beta}_1$-adrenoceptors were increased, but the level for ${\beta}_2$-adrenoceptors was decreased in the ADX rats. The mRNA levels for the other three subtypes and for the general and neuronal specific housekeeping genes, glyceroaldehyde-3-phosphate dehydrogenase (GAPDH) and N-enolase, respectively, were not changed in the ADX rats. In conclusion, the results indicate that adrenal steroids selectively regulate the gene expression of adrenoceptor subtypes in the PVN.

• Journal of Life Science
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• v.14 no.1
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• pp.121-125
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• 2004

The synergistic effect of lowered incubation temperature and CroEL/ES expression on the production of soluble form of B. macerans cyclodextrin glucanotransferase (CGTase) was studied in recombinant E. coli. pTCGTl and pGroll carrying the cgt and groEL/ES genes under the control of T7 promoter and pzt-I promoter, respectively, were co-introduced. Tetracycline (10 ng/ml) and IPTG (1 mM) were added at the early-exponential phase (2 hr) and mid-exponential phase (3 hr). Low temperature cultivation at $25^{\circ}C$ with groEL/ES expression improved the activity of CGTase by two fold, compared to $37^{\circ}C$ cultivation without chaperones. SDS-PACE analysis revealed that about 69% of CGTase in the total CGTase protein was found in the soluble fraction by overexpression of GroEL/ES and cultivation at$25^{\circ}C$, whereas 20% of CGTase was detected in the soluble fraction when E. coli was cultivated at $37^{\circ}C$ without chaperone. The amount of soluble CGTase from $25^{\circ}C$ culture with chaperone was 3.5-fold higher than that of $37^{\circ}C$ culture without chaperone. Therefore the expression of CroEL/ES and low temperature cultivation greatly enhanced the soluble production of CGTase in E. coli.

• Korean Journal of Plant Tissue Culture
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• v.26 no.3
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• pp.163-169
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• 1999

This study was conducted to produce herbicide resistant plants by transferring phosphinothricin acetyltransferase (PAT) gene into Populus alba $\times$ Populus glandulosa No .3 using Agrobacterium tumefaciens MP 90/PAT. Leaf segments from in vitro grown shoots of hybrid poplar No. 3 were soaked in a AB medium containing Agrobacterium tumefaciens MP 90/PAT for 10 min and cocultivated for 2 days on MS medium containing 1.0 mg/L 2,4-D and 0.2mg/L kinetin (CIM). Putative transformed calli could be selected after cocultivation of leaf segments on CIM supplemented with 50mg/L kanamycin and 500mg/L cefotaxime for 3 weeks. The selected calli were cultured on CIM supplemented with 50 mg/L kanamycin and 500 mg/L cefotaxime for 5~8 weeks before transfer to WPM containing 1.0mg/L zeatin, 0.1mg/L BAP, 50 mg/L kanamycin and 500mg/L cefotaxime for shoot regeneration. Shoots were regenerated from the callus after 4 week cultivation, and the regenerants were grown on the same medium for 7~l0 weeks. The plants rooted on 1/2 WPM containing 0.2 mg/L IBA and 50 mg/L kanamycin. To confirm the gene insertion into plants, GUS activity was detected by histochemical assay in the transformed plants. Finally, the presence of both NPT II and PAT genes from the transgenic plants were confirmed by PCR amplification with the gene specific primers and subsequent PCR-Southern blot with DIG-labeled PAT gene probe. After acclimatization in pots for 4 weeks, the plants were sprayed by 3 mL/L of Basta to test resistance to the herbicide. The transgenic plants remained green, whereas all the control plants died after one week.

• Korean Journal of Medicinal Crop Science
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• v.11 no.2
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• pp.161-166
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• 2003

Korean ginseng(Panax gmseng C.A. Meyer) is very difficult to obtain stable production of qualified ginseng roots because of variable stresses in soil environments. In environment stresses, soil condition is the most important factor, among which nutrients, especially inorganic materials such as N, P, K, Ca, Mg, Fe, etc., influence greatly on the ginseng growth. However, present ginseng field soils in Korea contain so much amount of such inorganic materials that a variety of remarkable disorders were noted in many ginseng plantations, resulting in decrease of qualitative ginseng production. Therefore, it is required to search for genetic resources and genes tolerant to salt stress for the development of ginseng cultivars. Selection of stress-tolerant ginseng lines in fields is very difficult because it is almost impossible to control properly the environmental conditions of soil. On the contrary, it can be studied with ease to search for stress-tolerant ginseng lines through in vitro culture because of easy manipulation of stress conditions. Murashige & Skoog(MS) media with 2.5 folds of $KNO_3,\;NH_4NO_3,\;MgSO_4\;7H_2O,\;KH_2PO_4,\;and\;CaCl_2\;2H_2O$ was established for the selection of ginseng lines tolerant to salt stress under the embryo culture.

• Korean Journal of Microbiology
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• v.44 no.3
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• pp.264-269
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• 2008

To develop an amylolytic industrial yeast strain producing $\beta$-amylase, the BAMY gene encoding Achlya bisexualis $\beta$-amylase was constitutively expressed under the control of the alcohol dehydrogenase gene promoter (ADC1p) in an industrial strain of Saccharomyces cerevisiae. Yeast transformation was carried out by an integration system containing $\delta$-sequences as the recombination site. The integrative cassette devoid of bacterial DNA sequences was constructed that contains the BAMY gene and $\delta$-sequences. Industrial S. cerevisiae transformed with this integrative cassette secreted 45 kDa $\beta$-amylase into the culture medium. The $\beta$-amylase activity of the transformant was approximately 18.5-times higher than that of A. bisexualis. The multi-integrated BAMY genes in the transform ant were stable after 100 generations of growth in nonselective medium. Hydrolysis of soluble starch and various starches with the enzyme released maltose but not glucose or oligosaccharides.

• Journal of Crop Science and Biotechnology
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• v.11 no.4
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• pp.269-276
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• 2008

Field resistance is defined as the resistance that allows effective control of a parasite under natural field condition and is durable when exposed to new races of that parasite. To identify the genes for field resistance to rice blast, quantitative trait loci (QTLs) conferring the resistance for races and blast nursery screening in japonica rice cultivars were detected and mapped using SSR markers. QTL analysis was carried out in 190 RILs population from the cross between Suweon365 (moderately resistant) and Chucheong (highly susceptible). Twelve QTLs against nine blast races inoculated were detected on chromosomes 1, 2, 4, 6, 7, 11 and 12. They explained from 5.1% to 34.9% of total phenotypic variation. Eight QTLs against blast nursery screening in four regions for three years were detected on chromosomes 1, 2, 4, 11 and 12. The phenotypic variation explained by each QTL ranged from 4.3% to 37.7%. Three chromosome segment substitution lines (CSSLs) of $BC_2F_6$ by backcross method were developed to transfer the QTLs into the susceptible cultivar Chucheong as a recurrent parent. A CSSL4-1 containing two QTLs qLB6.2 and qLB7 against blast races showed to the reaction of 6 to 7 at blast nursery in two regions for two years. The CSSL4-2 and CSSL93 containing QTLs, qLB11.2 and qLB12.1 of the resistance against leaf blast in blast nursery screening, respectively, had enhanced the resistance for blast nursery screening across two regions and in two years.

• Journal of Life Science
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• v.21 no.3
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• pp.358-367
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• 2011

Soybean is known to contain various phytochemicals that are related to anti-oxidant, anti-inflammatory and anti-obesity effects in mice and humans. The anti-obesity effect of by-product extracts from soybean on the differentiation of 3T3-L1 pre-adipocytes to adipocytes was investigated by suppressing adipocyte differentiation and lipid accumulation with Oil Red-O assay and quantitative PCR. In inducing differentiation of 3T3-L1 pre-adipocytes in the presence of an adipogenic cocktail, isobutylmethylanthine (IBMX), dexamathasone, and insulin, treatment with filtrated soybean soaked water, soybean milk, and soycurd residue from soybean curd processing significantly decreased mRNA expression of obesity-related gene such as PPAR${\gamma}$, Fabp4, and Scd1, adipsin, apolipoprotein (APOE) and adiponectin (ADIPOQ) without any significant cytotoxicity. We also determined the well-known isoflavones in soybean, such as daidzein and genistein, in the by-product extracts. Taken together, we suggest that soybean by-product extract showed anti-obesity effect by suppressing adipocyte related gene expression, and that by-products collected during soybean curd processing may be a good candidate as an ingredient in health care products.

• Journal of Life Science
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• v.21 no.7
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• pp.997-1003
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• 2011

Menopause and obesity are associated with metabolism. The purpose of this study was to examine the changes of PPAR${\gamma}$, PGC-1(${\alpha},\;{\beta}$), NRf-1 and TFAM mRNA and mitochondria biogenesis in adipocytes and investigate the effect of swimming exercise for 6weeks on ovariectomized rats. Rats were randomly assigned to 3 groups: (1) ovariectomized rats fed with a control diet (C, n=4), (2) ovariectomized rats fed with high fat diet (H, n=4), and (3) ovariectomized rats trained to exercise and fed with high fat diet (H+EX, n=4). Exercise was performed by swimming for 5 days/wk, with a progressive increase in exercise over the course of 6 weeks. Results showed that the fat tissue weight in the H group was markedly increased (p<0.01) compared to other groups, however, regular exercise significantly decreased fat weight. The PPAR-${\gamma}$ (p<0.05), PGC-$1{\alpha}$ (p<0.01), -$1{\beta}$ (p<0.05), NRf-1 (p<0.01) and TFAM (p<0.05) mRNA expression in the adipocytes of H+EX were higher than in the H group. These results suggest that regular exercise for 6 weeks might exert positive effects by increasing PPAR-${\gamma}$, PGC-1 (${\alpha},\;{\beta}$), NRf-1 and TFAM mRNA expression and mitochondria in adipocytes. Thus, regular exercise may be helpful in the improvement of mitochondria biogenesis function in obese, ovariectomized rats.