• Proceedings of the Botanical Society of Korea Conference
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• 1996.07a
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• pp.61-74
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• 1996

Human and monogastric animals can not synthesize 10 out of the 20 amino asids and therefor need to obtain these from their diet. The plant seed is a major source of dietary protein. It is particular important in their study to increase nutritional quality of the seed storage proteins. The low contents of lysine, asparagine and threonenein various cereal seeds and of cystein and methionine. In legume seeds is due to the low proportions of these amino acids in the major storage proteins, we have tried to apply the three strategies; (1) mutagenesis and selection of specific amino acid analogue resistance, (2) cloning and expression study of lysine biosynthesis related gene, (3) transfomation of lysine rich soybean glycinin gene. The 5-methyltryptophan (5MT) resistant cell lines, SAR1, SAR2 and SAR3 were selected from anther derived callus of rice (Oryza sativa L. "Sasanishiki"). Among these selected cell lines, two (SAR1 and SAR3) were able to grow stably at 200 mg/L of 5MT. Analysis of the freed amino acids in callus shows that 5MT resistant cells (SAR3) accumulated free tryptophan at least up to 50 times higher than those that of the higher than of SAS. These results indicated that the 5MT resistant cell lines are useful in studies of amino acid biosynthesis. Tr75, a rice (Oryza sativa L., var. Sasanishiki) mutant resistant to 5MT was segregated from the progenies of its initial mutant line, TR1. The 5MT resistant of TR75 was inherited in the M8 generations as a single dominant nuclear gene. The content of free amino acids in the TR75 homozygous seeds increased approximately 1.5 to 2.0 fold compared to wild-type seeds. Especially, the contents of tryptophan, phenylalanine and aspartic acid were 5.0, 5.3 and 2.7 times higher than those of wild-type seeds, respectively. The content of lysine is significantly low in rice. The lysine is synthesized by a complex pathway that is predominantly regulated by feedback inhibition of several enzymes including asparginase, aspatate kinase, dihydrodipicolinat synthase, etc. For understanding the regulation mechanism of lysine synthesis in rice, we try to clone the lysine biosynthetic metabolism related gene, DHPS and asparaginase, from rice. We have isolated a rice DHPS genomic clone which contains an ORF of 1044 nucleotides (347 amino acids, Mr. 38, 381 daltons), an intron of 587 nucleotides and 5'and 3'-flanking regions by screening of rice genomic DNA library. Deduced amino acid sequence of mature peptide domain of GDHPS clone is highly conserved in monocot and dicot plants whereas that of transit peptide domain is extremely different depending on plant specie. Southern blot analysis indicated that GDHPS is located two copy gene in rice genome. The transcripts of a rice GDHPS were expressed in leaves and roots but not detected in callus tissues. The transcription level of GDHPS is much higher in leaves indicating enormous chloroplast development than roots. Genomic DNA clones for asparaginase genes were screened from the rice genomic library by using plaque hybridization technique. Twelve different genomic clones were isolated from first and second screening, and 8 of 12 clones were analyzed by restriction patterns and identified by Southern Blotting, Restriction enzyme digestion patterns and Southern blot analysis of 8 clones show the different pattern for asparaginase gene. Genomic Southern blot analysis from rice were done. It is estimated that rice has at least 2-3 copy of asparaginase gene. One of 8 positive clones was subcloned into the pBluescript SK(+) vector, and was constructed the physical map. For transformation of lysine rich storage protein into tobacco, soybean glycinin genes are transformed into tobacco. To examine whether glycinin could be stably accumulated in endosperm tissue, the glycinin cDNA was transcriptionally fused to an endosperm-specific promotor of the rice storage protein glutelin gene and then introduced into tobacco genomic via Agrobacterium-mediated transformation. Consequently the glycinin gene was expressed in a seed-and developmentally-specific manner in transgenic tobacco seeds. Glycinin were targeted to vacuole-derived protein bodies in the endosperm tissue and highly accumulated in the matrix region of many transgenic plant (1-4% of total seed proteins). Synthesized glycinin was processed into mature form, and assembled into a hexamer in a similar manner as the glycinin in soybean seed. Modified glycinin, in which 4 contiguous methionine residues were inserted at the variable regions corresponding to the C - teminal regions of the acidic and basic polypeptides, were also found to be accumulated similarly as in the normal glycinin. There was no apparent difference in the expression level, processing and targeting to protein bodies, or accumulation level between normal and modified glycinin. glycinin.

• Food Science and Preservation
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• v.20 no.3
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• pp.323-329
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• 2013

Strawberries are among the leading ready-to-eat agricultural products that have superior taste and nutrition. Thus, consumer concerns about the safety of eating strawberries are growing. To evaluate the contamination levels of strawberries according to their cultivation methods (nutriculture, pesticide-free culture and organic farming) and parts [fruit (flesh), stalk (pedunle) and leaf (calyx)], 1,020 parts of strawberry samples were collected at 12 farms in Nonsan-si and quantitatively or qualitatively examined for the indicators of food safety and food poisoning bacteria. The total aerobic bacteria count in the whole samples was 2.3~6.8 ${\log}_{10}$ CFU/g, and coliform bacteria were detected in 14.2% of the whole samples with a contamination level range of 2.1~4.5 log CFU/g. E. coli were detected in 0.9% of the whole samples with a contamination level range of 2.1~2.8 log CFU/g. The analysis of the bacterial levels according to the cultivation methods showed that the total aerobic bacteria and coliform counts were higher in the strawberries that were grown via organic farming than in those that were grown via nutriculture and pesticide-free culture. However, the E. coli counts of the strawberries that were grown via organic farming and via pesticide-free culture were similar and differed from that of the strawberries that were grown via nutriculture. The analysis of the contamination levels according to the parts of the strawberries showed that the total aerobic bacteria, coliform and E. coli counts of the fruits, stalks and leaves of the strawberries did not significantly differ. Staphylococcus aureus was detected in two organically grown strawberries, but Salmonella spp., Listeria monocytogenes and E. coli O157:H7 were not detected in the whole samples. These results show that the bacterial contamination levels of the strawberries differed based on their cultivation methods. Thus, a suitable method of reducing the bacterial contamination levels of strawberries according to their farming methods is needed.

• The Korean Journal of Mycology
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• v.17 no.2
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• pp.82-90
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• 1989

Detection of antibody against pathogenic fungi in serum specimens of the patients with pulmonary tuberculosis or other lung diseases has been carried out(male) using the indirect fluorescence antibody technique and immunodiffusion tests. Immunodiffusion tests revealed that 104(36.5%) out of 285 patients examined showed a positive precipitin reaction against one or more of fungal antigens. The majority of ID positive patients 64(61.5%) reacted with Aspergillus fumigatus antigen and 49(47.1%) patients reacted with Candida albicans antigen ID positive reaction to A. fumigatus was found little more frequently among male patients, while Candida albicans reactors were found more frequently among female patients. Age distribution of ID positive reactors was high(49.1-43.3%) in age group of 40-59 years, but least or none in age group of less than 30 years. Age of fungal mycelium used as antigen did not effect sensitivity of the indirect flubrescence (IF) technique in detecting antibody to A. fumigatus. Antibody class against A. fumigatus that showed highest titer was IgG and thus FITC labeled anti-IgG immunoglobulin shoul be preferable. As relatively large amount of cell wall components of Aspergilli shared antigenically, a considerable cross-reaction was observed among A. fumigatus, A. flavus and A. niger, but not much with C. albicans. While (IF) has much better sensitivity when compared with ID, relative specificity of the latter procedure cannot to be overried, so that they could be batter used together in order to obtain quantitative measurement of antibody with relative specificity.

• 한국균학회소식:학술대회논문집
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• 2014.10a
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• pp.9-9
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• 2014

Null mutants generated by targeted gene replacement are frequently used to reveal function of the genes in fungi. However, targeted gene deletions may be difficult to obtain or it may not be applicable, such as in the case of redundant or lethal genes. Constitutive expression system could be an alternative to avoid these difficulties and to provide new platform in fungal functional genomics research. Here we developed a novel platform for functional analysis genes in Magnaporthe oryzae by constitutive expression under a strong promoter. Employing a binary vector (pGOF1), carrying $EF1{\beta}$ promoter, we generated a total of 4,432 transformants by Agrobacterium tumefaciens-mediated transformation. We have analyzed a subset of 54 transformants that have the vector inserted in the promoter region of individual genes, at distances ranging from 44 to 1,479 bp. These transformants showed increased transcript levels of the genes that are found immediately adjacent to the vector, compared to those of wild type. Ten transformants showed higher levels of expression relative to the wild type not only in mycelial stage but also during infection-related development. Two transformants that T-DNA was inserted in the promotor regions of putative lethal genes, MoRPT4 and MoDBP5, showed decreased conidiation and pathogenicity, respectively. We also characterized two transformants that T-DNA was inserted in functionally redundant genes encoding alpha-glucosidase and alpha-mannosidase. These transformants also showed decreased mycelial growth and pathogenicity, implying successful application of this platform in functional analysis of the genes. Our data also demonstrated that comparative phenotypic analysis under over-expression and suppression of gene expression could prove a highly efficient system for functional analysis of the genes. Our over-expressed transformants library would be a valuable resource for functional characterization of the redundant or lethal genes in M. oryzae and this system may be applicable in other fungi.

• Radiation Oncology Journal
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• v.26 no.4
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• pp.257-262
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• 2008

Purpose: Oral mucositis induced by radiotherapy to the head and neck area, is a common acute complication and is considered as the most severe symptom for cancer patients in the early stages of treatment. This study was proposed to establish the oral mucositis mouse model induced by a single dose of radiation for the facility of testing therapeutic candidates which can be used for the oral mucositis treatments. Materials and Methods: Fifty-five BALB/c mice were divided into four groups: control, 16 Gy, 18 Gy, and 20 Gy. Oral mucositis was induced by a single dose of radiation to the head and neck using 6 MV x-Ray from linear accelerator. After irradiation, body weight and physical abnormalities were checked daily. Tongue tissues from all groups were taken on days 1, 2, 3, 5, 7, 9, and 14, respectively and H&E staining was conducted to examine morphological changes. Results: Body weight dramatically decreased after day 5 in all irradiated mice. In the 16 Gy treatment group, body weight was recovered on day 14. The histology data showed that the thickness of the epithelial cell layer was decreased by the accumulated time after radiation treatment, up to day 9. Severe ulceration was revealed on day 9. Conclusion: A single dose of 16 Gy is sufficient dose to induce oral mucositis in Balb/C mice. Significant changes were observed in the Balb/C mice on days 7 and 9 after radiation. It is suggested that this mouse model might be a useful standard tool for studying oral mucositis induced by radiation.

• Journal of Life Science
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• v.20 no.3
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• pp.381-387
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• 2010

To find offspring of Jeju Black cattle (JBC) produced by embryo transfer (ET) and artificial insemination (AI), a molecular genetic study was carried out in candidate cattle populations collected from cattle farms in Jeju Island, Korea. The genetic marker set was composed of 11 ISAG microsatellite (MS) markers, 11 SAES MS markers selected by our preliminary analysis for population diversity of JBC and two major coat color related genes: MC1R and ASIP. The results showed a combined non-exclusion probability for first parent (NE-P1) that was higher than that recommended by ISAG (above 0.9995), and a combined non-exclusion probability for sib identity of $5.3{\times}10^{-10}$. Parentage analysis showed that the cases identified the candidate's father only (77.0%), mother only (54.0%), and both parents (40.5%) in the candidate offspring population. The ET and AI calves were identified as 14.7% in the in vitro fertilized eggs provided and 32.4% in total population, respectively. However, the result from ISAG marker analysis showed 3 identical allele-combinations in 7 calves, and that from ISAG/SAES MS marker combination also showed 1 identical allele-combination in 2 calves. Data from MS and coat-color gene analyses provided information for complete identification of all animals tested. Because the present JBC population was mostly bred using small nuclear founders through bioengineering techniques such as AI and ET, the genetic diversity levels obtained from MS analysis in the JBC population were relatively lower than those of other cattle populations, including Hanwoo. The results suggested that the more efficient marker combinations, including coat color related genotypes, should be studied and used for constructing a system for identification and molecular breeding of JBC as well.

• Korean journal of food and cookery science
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• v.22 no.6 s.96
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• pp.949-956
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• 2006

Eleven rice varieties, five normal-yielding and six high-yielding, were examined for proximate composition, mineral and fatty acid composition, and antinutrients. The proximate rice compositions for moisture, crude protein, crude fat, carbohydrate, crude fiber and crude ash were 10.70${\sim}$15.88%, 5.03${\sim}$7.73%, 0.45${\sim}$1.33%, 74.56${\sim}$82.05%, 0.30${\sim}$0.73% and 0.31${\sim}$0.91%, respectively. The proximate composition of the various rice varieties was not significantly different except for moisture and carbohydrate. The contents of saturated fatty acid, monounsaturated fatty acid and polyunsaturated fatty acid of the rice varieties were 22.45${\sim}$25.99%, 36.50${\sim}$44.99% and 32.01${\sim}$46.02%, respectively. Several high-yielding rices varieties had higher oleic acid or linoleic acid. Ilmi and Nampyung had high oleic acid contents at ever 40%, and Dongjin No. 1, IlPum, Junam and Seachuchung had high linoleic acid contents at over 37%. The major minerals of rice were K and Mg and their respective contents were 85.62% and 25.55%. High-yielding rice varieties had significantly higher Mn and Na contents and lower Mg and Fe contents than normal-yielding rice varieties (p<0.05). Antinutrients such as lectin, phytic acid and trypsin inhibitor were not detected in any of the rice varieties.

• Korean Journal of Agricultural and Forest Meteorology
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• v.1 no.1
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• pp.1-11
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• 1999

Rice yield and primary productivity (NPP) are dependent upon the variability of climate and soil. The variability and regionality of the rice yield and net primary productivity were evaluated with the meteorological data collected from Korea Meteorology Administration and the actual rice yield data from the Ministration of Agriculture and Forestry, Korea. The estimated NPP using the three models, dependent upon temperature(NPP-T), precipitation(NPP-P) and net radiation(NPP-R), ranged from 10.87 to 17.52 Mg ha$^{-1}$ with average of 14.69 Mg ha$^{-1}$ in the South Korea and was ranged 6.47 to 15.58 Mg ha$^{-1}$ with average of 12.59 Mg ha$^{-1}$ in the North Korea. The primary limiting factor of NPP in Korea was net radiation, and the secondary limiting factor was temperature. Spectral analysis on the long term change in air temperature in July and August showed periodicity. The short periodicity was 3 to 7 years and the long periodicity was 15 to 43 years. The coefficient of variances, CV, of the rice yield from 1989 to 1998 ranged 3.23 percents to 12.37 percents which were lower than past decades. The CV's in Kangwon and Kyeongbuk were high while that in Chonbuk was the lowest. The prediction model based on th e yield index and yield response to temperature obtain ed from the field crop situation showed reasonable results and thus the spatial distributions of rice yield and predicted yield could be expressed in the maps. The predicted yields was well fitted with the actual yield except Kyungbuk. For better prediction, modification should be made considering radiation factor in further development.

• The Korean Journal of Pharmacology
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• v.14 no.1_2
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• pp.33-40
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• 1978

1) The authors studied the effect of increasing age on the contraction and relaxation mechanism in the ureteral smooth muscle of the guinea pig. 2) Two to three week old, three month old, and two to three year old guinea pig ureters were used and the consistent amplitude of contratile responses were induced by using train stimulation. 3) After mounting the specimens in Tyrode's solution containing 2.6mM $Ca^{++}$, the ureter was stimulated, of which amplitude was initial contraction and next continuously superfused with $Ca^{++}$-free Tyrod's solution. When the contractile response stopped by electrical field stimulation, the muscle specimens was superfused with Tyrode's solution 0.25mM $Ca^{++}$ for 15min and stimulated with the same parameters. Thereafter, the contraction of $Ca^{++}$ in the solution was increased step by step up to 2.7mM. 4) The ureters of 2-3 week old guinea pigs needed less $Ca^{++}$ for the recovery of contractile response than those of three month and two to three year old did. In 2.7mM $Ca^{++}$, the ureters of 2-3 week and 3 month old guinea pigs recovered the contractile response of over 90% but those of 2-3 year old recovered the contractility of 77.2%. 5) Isoproterenol inhibited in dose dependent manner from $10^{-7}$ to $10^{-5}\;M$ ureteral contractility of both 2-3 week and 2-3 year old guinea pigs. The inhibition of the old ureter by isoproterenol was significantly less (P<0.025) than that of the younger ureter. However theophylline showed the strong inhibition independent of the function of age. 6) Dibutyryl cyclic AMP showed dose-dependent inhibition of the contraction of ureters of 2-3 week old guinea pigs but there was shown no inhibition in the old ureters. Further, the content of endogenous cyclic AMP in the two week old ureter was higher by 73% than that of 17 month old ureter. Cyclic GMP contents was not much different between two groups. 7) The ureteral smooth muscle of the younger guinea pig had more efficiency than that of the older animals in the mobilization and storage of calcium which concerned itself in the contraction and relaxation mechanism.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.24 no.3
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• pp.203-213
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• 1991

Zooplankton samples were collected vertically from different layers with a closing net at 14 stations in the middle East Sea of Korea in February, August and September to study distribution of biological indicators for analysis of water masses. Horizontal and vertical distributions of important species of copepods and chaetognathas known as indicator species were closely related to distributions of different water masses and oceanic fronts. Pleuromamma gracilis, Calanus tenuicornis, Sagitta enflata and Sagitta minima were found to be reliable indicator species to determine warm water mass with warm core, and Calanus cristatus, Calanus tonsus and Sagitta elegans could be used as cold water species for evaluating the movement of cold current from North Korea, and Gaetanus armiger was deep sea water species. Therefore, it was found that North Korean Cold Current down to the south along the coast appeared to be significant in the surface around Chumunjin area, and from here towards the south the cold water containing S. elegans submerged under warm water with S. enflata which were about $2{\~}4^{\circ}C$ higher than that of the vicinity and reappeared near Chukpeon area in surface layer. In the layer between loom and 300m depths, distribution of Pleuromamma gracilis and Sagitta bedoti indicated that warm water mass and front zone influenced by the different water systems were formed in northwestern area off Ulreung-do. In $300{\~}500m$ layer, the proper cold water could be estimated to be present in the northwestern area off Ulreung-do throughout the survey period by the high abundance of Gaetanus armiger. In August, distributions of S. bedoti, S. enflata and S. minima were valuable index to find oceanic fronts and warm core.