• Title/Summary/Keyword: Culture Conditions

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Species-specific Expression of Rpia Transcript in Cumulus-oocyte-complex (난자-난구세포 복합체에서 발현하는 Rpia 유전자의 종 특이적 발현)

  • Kim, Yun-Sun;Yoon, Se-Jin;Kim, Eun-Young;Lee, Kyung-Ah
    • Clinical and Experimental Reproductive Medicine
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    • v.34 no.2
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    • pp.95-106
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    • 2007
  • Objective: We previously identified differentially expressed genes (DEGs) between germinal vesicle (GV) and metaphase II (MII) mouse oocyte. The present study was accomplished as a preliminary study to elucidate the role of ribose 5-phosphate isomerase A (Rpia), the essential enzyme of the pentose phosphate pathway (PPP), in oocyte maturation. We observed expression of Rpia in the mouse and porcine oocytes. Methods: Expression pattern of the 11 MII-selective DEGs in various tissues was evaluated using RT-PCR and selected 4 genes highly expressed in the ovary. According to the oocyte-selective expression profile, we selected Rpia as a target for this study. We identified the porcine Rpia sequence using EST clustering technique, since it is not yet registered in public databases. Results: The extended porcine Rpia nucleotide sequence was submitted and registered to GenBank (accession number EF213106). We prepared primers for porcine Rpia according to this sequence. In contrast to the oocyte-specific expression in the mouse, Rpia was expressed in porcine cumulus and granulosa cells as well as in oocytes. Conclusion: This is the first report on the characterization of the Rpia gene in the mouse and porcine ovarian cells. Results of the present study suggest that the mouse and porcine COCs employ different mechanism of glucose metabolism. Therefore, the different metabolic pathways during in vitro oocyte maturation (IVM) in different species may lead different maturation rates. It is required to study further regarding the role of Rpia in glucose metabolism of oocytes and follicular cell fore exploring the regulatory mechanism of oocyte maturation as well as for finding the finest culture conditions for in vitro maturation.

Characterization and Purification of the Bacteriocin Produced by Bacillus licheniformis Isolated from Soybean Sauce (간장에서 분리한 Bacillus licheniformis가 생산하는 박테리오신의 특성 및 정제)

  • Jung, Sung-Sub;Choi, Jung-I;Joo, Woo-Hong;Suh, Hyun-Hyo;Na, Ae-Sil;Cho, Yong-Kweon;Moon, Ja-Young;Ha, Kwon-Chul;Paik, Do-Hyeon;Kang, Dae-Ook
    • Journal of Life Science
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    • v.19 no.7
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    • pp.994-1002
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    • 2009
  • A bacteriocin-producing bacterium identified as Bacillus licheniformis was isolated from soybean sauce. Antibacterial activity was confirmed by paper disc diffusion method, using Micrococcus luteus as a test organism. The bacteriocin also showed antibacterial activities against Bacillus sphaericus, Lactobacillus bulgaricus, Lactobacillus planiarum, Paenibacillus polymyxa, and Pediococcus dextrinicus. Optimal culture conditions for the production of bacteriocin was attained by growing the cells in an MRS medium at a pH of 6.5~ 7.0 and a temperature of 37$^\circ$C for 36$\sim$48 hr. Solvents such as chloroform, ethanol, acetone, and acetonitrile had little effect on bacteriocin activity. However, about 50% of bacteriocin activity diminished with treatment of methanol and isopropanol at the final concentration of 50% at 25$^\circ$C for 1 hr. It was stable against a pH variation range from 3.0 and 7.0, but the activity reduced to 50% at a pH range from 9.0 to 11.0. It's activity was not affected by heat treatment at 100$^\circ$C for 30 min and 50% of activity was retained after heat treatment at 100$^\circ$C for 60 min, showing high thermostability. The bacteriocin was purified to a homogeneity through ammonium sulfate precipitation, SP-Sepharose ion-exchange chromatography, and reverse-phase high-performance liquid chromatography (HPLC). The entire purification protocol led to a 75-fold increase in specific activity and a 13.5% yield of bacteriocin activity. The molecular weight of purified bacteriocin was estimated to be about 2.5 kDa by tricine-SDS-PAGE.

Isolation and Characteristics of a Phenol-degrading Bacterium, Rhodococcus pyridinovorans P21 (페놀분해세균 Rhodococcus pyridinovorans P21의 분리 및 페놀분해 특성)

  • Cho, Kwang-Sik;Lee, Sang-Mee;Shin, Myung-Jae;Park, Soo-Yun;Lee, Ye-Ram;Jang, Eun-Young;Son, Hong-Joo
    • Journal of Life Science
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    • v.24 no.9
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    • pp.988-994
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    • 2014
  • The effluents of chemical and petroleum industries often contain non-biodegradable aromatic compounds, with phenol being one of the major organic pollutants present among a wide variety of highly toxic organic chemicals. Phenol is toxic upon ingestion, contact, or inhalation, and it is lethal to fish even at concentrations as low as 0.005 ppm. Phenol biodegradation has been studied in detail using bacterial strains. However, these microorganisms suffer from substrate inhibition at high concentrations of phenol, whereby growth is inhibited. A phenol-degrading bacterium, P21, was isolated from oil-contaminated soil. The phenotypic characteristics and a phylogenetic analysis indicated the close relationship of strain P21 to Rhodococcus pyridinovorans. Phenol biodegradation by strain P21 was studied under shaking condition. The optimal conditions for phenol biodegradation by strain P21 were 0.09% $KNO_3$, 0.1% $K_2HPO_4$, 0.3% $NaH_2PO_4$, 0.015% $MgSO_4{\cdot}7H_2O$, 0.001% $FeSO_4{\cdot}7H_2O$, initial pH 9, and $20-30^{\circ}C$, respectively. When 1,000 ppm of phenol was added to the optimal medium, the strain P21 completely degraded it within two days. Rhodococcus pyridinovorans P21 could grow in up to 1,500 ppm of phenol as the sole carbon source in a batch culture, but it could not grow in a medium containing above 2,000 ppm. Moreover, strain P21 could utilize toxic compounds, such as toluene, xylene, and hexane, as a sole carbon source. However, no growth was detected on chloroform.

Enhanced Production of Oleanolic Acid by the Elicitation in Oldenlandia diffusa Suspension Cell Cultures (백화사설초의 현탁세포배양에서 Elicitation에 의한 Oleanolic acid 생산성 증대)

  • Lee Yong-Il;Kim Dong-Il
    • KSBB Journal
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    • v.19 no.6 s.89
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    • pp.471-477
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    • 2004
  • Oldenlandia diffusa is a Chinese medicinal herb with antitumor activity capable of suppressing the growth of some cancer cell lines. Oleanolic acid and ursolic acid are triterpenoid compounds that exist in Oldenlandia diffusa. Recently, these have been noted for anti-inflammatory, anti-cancer, and hepato-protective effects. Application of both plant growth regulators, 2,4-D and kinetin, was found to be essential for the initiation of callus and suspension cells. Leaf blades of Oldenlandia diffusa was transformed into callus on Schenk and Hildebrandt medium supplemented with 0.5 mg/L 2,4-D and 0.1 mg/L kinetin, while optimum initiation condition for suspension cells of Oldenlandia diffusa was determined to be 0.75 mg/L 2,4-D and 0.1 mg/L kinetin. Chromatographic separation of oleanolic acid from its derivatives was achieved using Rexchrom S5-100-ODS column. Analytical conditions for oleanolic acid were determined as follows: flow rate at 1.0 mL/min, UV length at 200 nm and mobile phase of $80\%$ acetonitrile and $20\%$ water. Production of secondary metabolites was found to be increased by the treatment with elicitors or signal transducers. The maximum production of oleanolic acid was 99.6 mg/L in cultures with 0.5 mM salicylic acid. It is 1.74 times higher than that of control.

Expression and Purification of Recombinant Human Interferon-gamma Produced by Escherichia coli (대장균이 생산한 재조합 인체 감마인터페론의 발현과 정제)

  • Park, Jung-Ryeol;Kim, Sung-Woo;Kim, Jae-Bum;Jung, Woo-Hyuk;Han, Myung-Wan;Jo, Young-Bae;Jung, Joon-Ki
    • KSBB Journal
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    • v.21 no.3
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    • pp.204-211
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    • 2006
  • For the production of the recombinant human interferon-gamma(rhIFN-${\gamma}$) in Escherichia coli, human glucagon and ferritin heavy chain were used as fusion partners. Even though rhIFN-${\gamma}$ is expressed as an inclusion body form in E. coli because of strong hydrophobicity of itself, over 50% of fused rhIFN-${\gamma}$ was expressed as soluble form in E. coli $Origami^{TM}$(DE3) harboring pT7FH(HE)-IFN-${\gamma}$ which encodes ferritin heavy chain-fused rhIFN-${\gamma}$. In the case of using glucagon-ferritin heavy chain hybrid mutant as a fusion partner, 6X His-tag was additionally introduced to N-terminus of GFHM(HE)-IFN-${\gamma}$ for enhancing purification yields of rhIFN-${\gamma}$. Fusion protein HGFHM(HE)-IFN-${\gamma}$ with two 6X His-tag was more effectively bound to Ni-NTA agarose bead than GFHM(HE)-IFN-${\gamma}$ with a 6X His-tag. rhIFN-${\gamma}$ was completely purified from enterokinase-treated HGFHM(HE)-IFN-${\gamma}$ by Ni-NTA affinity column. For high-level production of rhIFN-${\gamma}$, glucose was used as the sole carbon source with simple exponential feeding rate($2.4{\sim}7.2g/h$) in fed-batch process. The effective lactose concentration for the expression of the rhIFN-${\gamma}$ was $10{\sim}20mM$. Under the fed-batch culture conditions, rhIFN-${\gamma}$ production yield reached 11 g DCW/L for 6 hours after lactose induction.

Ecological Importance of Water Budget and Synergistic Effects of Water Stress of Plants due to Air Pollution and Soil Acidification in Korea (한국에서 수분수지의 생태적 중요성과 대기오염 및 토양 산성화로 인한 식물의 수분스트레스 증대 효과)

  • 이창석;이안나
    • The Korean Journal of Ecology
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    • v.26 no.3
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    • pp.143-150
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    • 2003
  • Korea has plentiful precipitation but rainfall events concentrate on several months of rainy season in her weather condition. Korea, therefore, experiences drought for a given period every year. Moreover the soil has usually low water holding capacity, as it is composed coarse particles originated from the granite. Response of several oaks and the Korean red pine (Pinus densiflora) on water stress showed that water budget was significant factor determining vegetation distribution. In addition, dehydration level due to cold resistance mechanism of several evergreen plants during the winter season was closely related to their distribution in natural condition. Experimental result under water stress showed that the Korean red pine was very tolerant to desiccation but the seedlings showed high mortality during the dry season. The mortality tended to proportionate to soil moisture content of each site. A comparison between soil moisture content during June when it is severe dry season and moisture content of the culture soil when the pine seedlings reached the permanent wilting point due to water withheld proved that high mortality during the dry season was due to water deficit. Water potential of sample plants measured during the exposure experiment to the air pollutant showed a probability that water related factors would dominate the occurrence of visible damage and the tolerance level of sample plants. In both field survey and laboratory experiment, plants exposed to air pollution showed more rapid transpiration than those grown in the unpolluted condition. The result would due to injury of leaf surface by air pollutants. Aluminum (Al/sup 3+/) increased in the acid soil not only inhibits root growth but also leads to abnormal distribution of root system and thereby caused water stress. The water stresses due to air pollution and soil acidification showed a possibility that they play dominating roles in inducing forest decline additionally to the existing water deficit due to weather and soil conditions in Korea. Sludge, which can contribute to improve field capacity, as it is almost composed of organic matter, showed an effect ameliorating the retarded growth of plant in the acidified soil. The effect was not less than that of dolomite known in widely as such a soil ameliorator. Litter extract contributed also to mitigate the water stress due to toxic Al/sup 3+/. We prepared a model showing the potential interaction of multiple stresses, which can cause forest decline in Korea by synthesizing those results. Furthermore, we suggested restoration plans, which can mitigate such forest decline in terms of soil amelioration and vegetation restoration.

A Study on Gardening Theories of Li, Yu in 'Xian Qing Ou Ji' (한정우기(閑情偶寄)를 통해 본 이어(李漁)의 조원(造園) 이론)

  • Zhang, Lin;Jung, Woo-Jin;Sung, Jong-Sang
    • Journal of the Korean Institute of Traditional Landscape Architecture
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    • v.36 no.3
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    • pp.137-148
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    • 2018
  • Li Yu's Xian Qing Ou Ji was written during the late Ming and early Qing Dynasty when material culture flourished due to the development of the commercial economy. Xian Qing Ou Ji is considered a retrospective work that reflects the rich material society at that time. This study focuses on his gardening theories through the interpretation and analysis of these two texts of 'Jushibu(居室部)' and 'Zhongzhibu(種植部)' in Xian Qing Ou Ji. The results are summarized as follows. First, he explained the method of garden configuration and orientation according to the topography utilizing the theory of 'suiting one's measures to local conditions(因地制宜)'. He emphasized the detailed understanding of topography as the starting point for all the gardening theories. It stated that he tried to adjust the height of the terrain artificially to create an ideal form of 'anterior high and posterior low(前低後高)'. The successful arrangement of houses, pavilions, rockery and waterways et al. was able to be achieved because of his accurate understanding of topography. Second, 'borrowing scenery(取景在借)' means 'borrowing the outside scenery to the inside'. He applied this theory by 'viewing in motion(動觀)' and 'viewing in repose(靜觀)'. 'Viewing in motion' is seen as a positive methodology for landscape enjoyment. For example, the view through a window changes with movements of the boat, and it can portray thousands of landscape paintings which can be seen through the windows. In the case of 'viewing in repose', the window is regarded as a means of attracting outside garden views to the inside. Third, he emphasized 'the firmness of objects(制體宜堅)', where the design of windows and railings must be considered ahead of others. Fourth, he interpreted the contents of 'the pursuit of novelty(創異標新)'. This generalizes the characteristics of his gardening theory. The contents included 'hall wall(廳壁)', 'variable sun visor(活?)', 'hanging boards and couplets(聯?)' and 'method of supporting vines with bamboo screen(竹屛扶植法)'.

The Historical Geography of Land-Use and Agriculture Along the Lower Nam-River Floodplains (남강 하류 범람원의 토지이용과 농업형태 변화에 관한 연구)

  • Lee, Jeon;Son, Ill
    • Journal of the Korean association of regional geographers
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    • v.4 no.2
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    • pp.31-47
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    • 1998
  • This paper deals with the historical geography of land-use and agriculture along the Lower Nam-River floodplains. The reclamation process of the river floodplains, the cultivation methods on the reclaimed lands, and the land-use patterns and processes are investigated. The Nam River, one of the major tributaries of the Nakdong River, flows through the boundary between Ham-An and Eu-Ryong Guns. Larger floodplains are located in Ham-An Gun. The floodplains of Ham-An Gun have been surveyed intensively in this study. In South Korea, the alluvial plains, mostly located along the river valleys, have been reclaimed to provide fertile agricultural lands. Those along the upper river valleys were reclaimed before those along the lower river valleys. The flood-plains of Han-An Gun were reclaimed to be the largest agricultural lands of the Gun. The natural levees along the Lower Nam-River Valley were identified before the reclamation processes but now hardly identified. Relatively larger floodplains are located along the tributary streams of the Nam River. Often there are low-lying back swamps between the natural levees and the hills/mountains that rise above the floodplains. The back swamps, called 'natural bog lands' in this region, have been reduced in size and in number through reclamation for the purpose of agricultural and industrial land-uses. Now about ten 'natural bog lands' are found in the Ham-An floodplains, and some of them are being reclaimed for the industrial land-use. This study suggests the emergent need of conservation for the remaining 'natural bog lands' in terms of ecology. Seven agricultural fields of large size, originated from the Nam-River floodplains, are identified in this study: Kun(큰들), Chung-Am(정암들), Chang-chi(장지들), Baek-San(백산들), Ha-Ki(하기들), Gu-Hae(구혜들), and Chang-Po(장포들) fields. The Kun field was reclaimed during the Japanese control and the Gu-Hae, in the 1950s. All of those except the above two fields were reclaimed after the mid-1960s. The Nam-River Dam in Chinju, completed in 1969, contributed the reclamation processes along the Lower Nam-River floodplains. The rice acreage of the region has been reduced slowly since 1970 but the rice production of the region has been relatively stable (Table 4). Rice culture had been the most important agriculture on the reclaimed lands for decades before the greenhouse vegetable cultivation became more important in the 1980s. Among the vegetables cultivated in the greenhouse, the watermelon is the dominantly leading one. Watermelons are usually harvested two or three times in a year though it is possible to harvest four times in one year. The rotation of watermelons and rice is common in the region. It is known the physical conditions of the Nam-River floodplains in Ham-An Gun is the most suitable for watermelon cultivation in South Korea.

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Effect of TGF-${\beta}$ Supplementation on In Vitro Maturation of Hanwoo COCs (Cumulus Oocytes Complexes) (TGF- ${\beta}$ 첨가가 한우 난포란의 체외성숙에 미치는 영향)

  • Choi, Sun-Ho;Lee, Hye-Hyun;Yeon, Seong-Heum;Han, Man-Hye;Kim, Hyun-Jong;Cho, Sang-Rae;Woo, Jae-Seok;Baek, Kwang-Soo;Ryu, Il-Sun;Son, Dong-Soo
    • Development and Reproduction
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    • v.8 no.2
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    • pp.119-122
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    • 2004
  • It is well known that unidentified factors in sera, hormones and growth factors promote the proliferation of granulosa cells and nuclear maturation of bovine COCs in vitro. Attemps had been developed the simple composition of culture media and similar system to in vivo conditions has been applied. In the present study, we investigated the effect of TGF-${\beta}$ on in vitro maturation and in vitro development of Hanwoo COCs. When the COCs were matured in TCM 199 containing 0.1, 1 or 10 ng/ml TGF-${\beta}$ for 24 hrs, metaphaseⅡ of COCs were obtained 95.8%, 100% of matured COCs, respectively and there were no differences among the concentrations of TGF-${\beta}$. Matured COCs with TGF-${\beta}$ cultured in maturation medium after in vitro fertilization, developmental rate to blastocyst were 0~0.8%. Matured COCs with TGF-${\beta}$ were cultured in TCM 199+10% FBS, 0.8% BSA, 0.1% PVA, blastocyst formation were showed in 12.4%, 12.8%, 8.5% of those and cultured in IVMD or IVD without serum were 38.4%, 34.8%, respectively. There were significant differences among the media (P<0.05). TGF-${\beta}$ is available for i vitro maturation of bovine COCs, but further investigation would be need for finding the synergistic autocrine/paracrine fashion of other growth factors in early bovine development.

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Studies on the Enzyme of Rhizopus oryzae - Part I. Production of Acid Protease and Enzymatic Characteristics - (Rhizopus oryzae의 효소(酵素)에 관(關)한 연구(硏究) - 제(第)1보(報), 산성(酸性) Protease 생산(生産) 및 효소(酵素)의 특성(特性) -)

  • Hou, Won-Nyong;Chung, Man-Jae
    • Applied Biological Chemistry
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    • v.22 no.3
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    • pp.135-141
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    • 1979
  • These experiments were conducted to investigate the conditions of the acid protease production by Rhizopus oryzae and the characteristics of crude enzyme. The results obtained were as follows: 1. The optimum culture time and the optimum amount of added water to the wheat bran medium were about 48 hrs and $80{\sim}120%$, respectively. 2. The addition of $(NH_4)_6Mo_7O_{24},\;(NH_4)_2SO_4,\;NH_4NO_3$, casein, and albumin, respectively, as nitrogen sources to the wheat bran medium was effective. Of these, the optimum concentrations of addition of $(NH_4)_6Mo_7O_{24}$ and casein which were the most effective were 0.1% and 1.0%, respectively. 3. The addition of glucose, galactose, maltose, lactose, and soluble starch, respectively, as carbon sources to the wheat bran medium was effective. Of these, the optimum concentration of addition of glucose which was the most effective was 3.0%. 4. The addition of $KH_2PO_4$ as a phosphate salt to the wheat bran medium was effective. The optimum concentration of addition of $KH_2PO_4$ was 0.3%. 5. The optimum pH for the enzyme action was 2.4, the optimum temperature about $40^{\circ}C$, and the stable pH range $2.0{\sim}5.0$. The enzyme was stab1e below $40^{\circ}C$. 6. The enzyme activity increased rapidly for 10 minutes after addition, thereafter it increased slowly. 7. The enzyme activity increased rapidly to 2 ml of addition, but nearly did not increase at the amounts greater than 2ml.

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