• Title/Summary/Keyword: plant cultured cells

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Disinfection Efficiency of Medium Pressure UV Lamp on Major Bacteria in Sand Filtered Water (사여과수에 존재하는 우점세균의 중압 자외선 램프 소독능)

  • Ahn, Seoung-Koo;Yang, Yoon-Yong
    • Journal of Korean Society of Environmental Engineers
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    • v.32 no.12
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    • pp.1141-1146
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    • 2010
  • Isolated the heterotrophic aerobic bacteria in sandfiltered water on NA and TSBA solid medium, selected 8 dominant species and identified by Sherlock System. Each samples are irradiated 0, 5, 16, 40 and $60\;mJ/cm^2$ using on CBD (Collimated Beam Device) Medium Pressure UV lamp after these identified bacterium did liquid culture how to make $10^6{\sim}10^7\;cells/mL$ suspended in dilution water. Then cultured bacteria are estimated inactivation rate on plate media. Identified Gram positive group are Bacillus Subtilus, Bacillus megaterium, Rhodococcus erythropolis and Microbacterium laevaniformans; Gram negative group are Pseudomonas vesicularis, Pseudomonas pseudoflava, Alcaligenes paradoxus and Zooglea ramigera. These isolation of bacterium are more stronger reference strain and high resistance of MP UV irradiation, Besides Gram negative bacterium are more sensitive Gram positive bacterium on MP UV dose. Now we are estimating to $60{\sim}100\;mJ/cm^2$ MP UV dose for efficient disinfection in water treatment plant.

Effect of ganglioside GD3 synthase gene expression on VSMC proliferation via ERK1/2 pathway, cell cycle progression and MMP-9 expression

  • Lee, Young-Choon;Kim, Cheorl-Ho
    • Proceedings of the Korean Society of Plant Biotechnology Conference
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    • 2004.10a
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    • pp.81-90
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    • 2004
  • Sialic acid containing glycosphingolipids (gangliosides) have been implicated in the regulation of various biological phenomena such as atherosclerosis. Recent report suggeststhat exogenously supplied disialoganglioside (GD3) serves a dual role in vascular smooth muscle cells (VSMC) proliferation and apoptosis. However, the role of the GD3 synthase gene in VSMC responses has not yet been elucidated. To determine whether a ganglioside is able to modulate VSMC growth. the effect of overexpression of the GD3 synthase gene on DNA synthesis was examined. The results show that the overexpression of this gene has a potent inhibitory effect on DNA synthesis and ERK phosphorylation in cultured VSMC in the presence of PDGF. The suppression of the GD3 synthase gene was correlated with the down-regulation of cyclinE/CDK2. the up-regulation of the CDK inhibitor p21 and blocking of the p27 inhibition,whereas up-regulation of p53 as the result of GD3 synthase gene expression was not observed. Consistently, blockade of GD3 function with anti-GD3 antibody reversed VSMC proliferation and cell cycle proteins. The expression of the CD3 synthase gene also led to the inhibition of TNF--induced matrix metalloproteinase-9 (MMP-9) expression in VSMC as determined by zymography and immunoblot. Furthermore, GD3 synthase gene expression strongly decreased MMP-9 promoteractivlty in response to TNF-. This inhibition was characterized by the down-regulation of MMP-9,which was Iranscriptionally regulated at NF-B and activation protein-1 (AP-1) sites in the MMP-9promoter Finally, the overexpression of MMP-9 in GD3 synthase transfectant cells rescued VSMC proliferation. However MMP-2 overexpression was not affected the cell proliferation. These findings suggest that the fl13 synthase gene represents a physiological modulator of VSMC responses that may contribute to plaque instability in atherosclerosis.

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Production of Green Fluorescent Protein (GFP) from Transgenic Rice Cell Suspension Culture (형질전환된 벼세포배양에서 green fluorescent protein (GFP) 생산)

  • Lee, Jae-Hwa
    • Journal of Life Science
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    • v.17 no.2 s.82
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    • pp.293-297
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    • 2007
  • Green fluorescent protein (GFP) is an attractive reporter for bioprocess monitoring. A fluorescence-based method was developed to quantify GFP levels in transgenic plants and protein extracts. In this study, GFP was produced and secreted from suspension cells derived from transgenic rice. The RAmy3E promoter placed before the GFP gene controlled by sugars such as sucrose. The effects of sucrose concentration on the secretion of GFP and total protein into the medium were investigated in batch suspension culture. It was possible, therefore, to induce the expression of the GFP by removing sucrose from the cultured media or by allowing the rice suspension cells to deplete sucrose catabolically. The dry cell weight (7.06 g/L) and GFP level were detected as highest at 12%, 3% sucrose after 20 day culture, respectively. However secreted GFP fluorescence at the other sucrose concentrations (6%, 12%, 18% and 24%) were a little amount in media.

Molecular Characterization of Three cDNA Clones Encoding Calmodulin Isoforms of Rice

  • Lee, Sung-Ho;Kim, Cha Young;Lim, Chae Oh;Lee, Soo In;Gal, Sang Wan;Choi, Young Ju
    • Journal of Applied Biological Chemistry
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    • v.43 no.1
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    • pp.5-11
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    • 2000
  • Three cDNA clones encoding rice calmodulin (CaM) isoforms (OsCaM-1, OsCaM-2, and OsCaM-3) were isolated from a rice cDNA library constructed from suspension-cultured rice cells treated with fungal elicitor. The coding regions of OsCaM-1 and O.sCaM-2 were 89% homologous at DNA Ievel, whereas the 5' and 3' untranslated regions were highly divergent. The polypeptides encoded by OsCaM-1 and OsCaM-2 was identical except two conservative substitution at position 8 and 75. The coding region of OsCaM-3 was consist of a typical conserved CaM domain and an additional C-terminal extension. The amino acid sequence of conserved CaM domain of OsCaM-3 shared only 86% identity with that OsCaM-1. The OsCaM-3 cDNA is belongs to a novel group of calmodulin gene due to its C-terminal extension of 38 amino acids, a large number of which are positively charged. The extension also contains a C-terminal CaaX-box prenylation site (CVlL). Genomic Southern analysis revealed at least six copies of CaM or CaM-related genes, suggesting that calmodulin may be represented by a small multigene family in the rice geneme. Expression of OsCaM gene was examined through Northern blot analysis. Transcript level of OsCaM-3 was increased by treatment with a fungal elicitor, whereas the OsCaM-1 and OsCaM-2 genes did not respond to the fungal elicitor. The expression of OsCaM-3 gene was remarkable inhibited in the rice cells treated with cyclosporine A, calcinurin inhibitor.

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Effects of Banha Extract on the Melanin Biosynthesis and Tyrosinase mRNA Level in Bl6 Mouse Melanoma Cells (반하 추출물이 B-16 마우스 흑색종 세포의 멜라닌 생성과 타이로시네이즈 mRNA 양에 미치는 영향)

  • 이상화;김진준
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.23 no.2
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    • pp.23-32
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    • 1997
  • Melanin pigmentation in human skin is a major defense mechanism against ultraviolet light of the sun. Tyrosinase(EC 1.14.18.1) plays a key role in the biosynthesis of ultraviolet of melanin. This is why much researches have been focused on its regulation in controlling the epidermal melanization. We have found that the water-extract of Banha(Pinelliae ternate B.), an oriental medicinal plant, has no tyrosinase inhibitory activity, but does inhibit the melanin biolsynthesis in B16 mouse melanin cells. We also found that Banha extract lowers the tyrosinase activity in cultured cells. To elucidate the action mechanism of Banha extract we have investigated its effect on the tyrosinase mRNA level using reverse transcription-polymerase chain reaction technique. It was revealed that Banha extract reduced the tyrosinase mRNA level in dose dependent manner; when B16 mouse melanoma cells were cultured with 2mg/ml and 5mg/ml of Banha extract, there were 20% and 44% decrease in tyrosinase mRNA level, respectively. These data suggest that the Banha extract exerts its melanogenic inhibitory effect through the transcriptional regulation of tyrosinase mRNA.

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Molecular Cloning and Expression of the Metallothionein Gene under Environmental Stresses in Sweet Potato (고구마 metallothionein 유전자의 클로닝 및 환경 스트레스 하에서 발현 분석)

  • Kim, Young-Hwa;Yu, Eun Jeong;Huh, Gyung-Hye
    • Journal of Life Science
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    • v.27 no.12
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    • pp.1415-1420
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    • 2017
  • The metallothionein (MT) gene (IbMT3) was selected from an EST library of suspension-cultured sweet potato cells. The MT gene, which is one of abundant ESTs in the library, is involved in stress regulation of cells and tissues. A full-length IbMT3 cDNA was obtained and analysis of its nucleotide sequence revealed that IbMT3 encoded a type 3 MT protein, based on its structural characteristics. The function of type 3 MT in plants is not yet known. Northern blot analysis showed stronger expression of IbMT3 in suspension-cultured cells than in sweet potato plant leaves. Since cell culture is known to impose a state of oxidative stress on cells, sweet potato plants were subjected to oxidative stress to investigate the transcriptional regulation of IbMT3. When the herbicide methyl viologen (MV) was administered for 6, 12, and 24 hr, IbMT3 transcription rapidly increased at 6 hr and then decreased. A cold treatment at $15^{\circ}C$ for 24 and 48 hr resulted in a gradual increase in IbMT3 expression. These findings indicate that IbMT3 expression is regulated in response to environmental and oxidative stress. IbMT3 isoform is expected to have antioxidant effects in sweet potato plants and may play an important role in cellular adaptation to oxidative stress.

Inhibitory Effect of Taraxci Herba Methanol Extract on Pro-inflammatory Mediator in Lipopolysaccharide;Activated Raw 264.7 cells

  • Jo, Mi-Jeong;Chu, Yan-Hui;Back, Young-Doo;Lee, Byung-Wook;Shin, Soon-Shik;Kwon, Young-Kyu;Kim, Sang-Chan
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.22 no.4
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    • pp.907-913
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    • 2008
  • Taraxci Herba (TH; Pogongyoung in Korean) has been used in traditional oriental medicine for the treatment of various ailments. The biological activity of this plant is not yet evaluated systematically. This study was conducted to evaluate the inhibitory effects of TH on the production of nitric oxide (NO) and the expression of inducible nitric oxide synthase (iNOS) in lipopolysaccharide (LPS)-activated Raw264.7 cells. The aim of the present work is to investigate a potential anti-inflammatory activity of TH. The Raw264.7 cells were cultured in DMEM medium for 24 h. After serum starvation for 12 h, the cells were treated with TH for 1 h, followed by stimulating NO production with LPS ($2{\mu}g/ml$). As result of this study, TH inhibited the levels of NO, PGE2, $TNF-{\alpha}$, IL-6 and $IL-1{\beta}$, and the expressions of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) activated by LPS. These inhibitory effects were mediated though the inhibition of phosphorylation of inhibitory kappa B ($I{\kappa}B$). These findings showed that TH could have some anti-inflammatory effects.

The Production of Anti-cancer Substances by in vitro Grown Cultures of Panax ginseng C.A. Meyer

  • Yang, Deok-Chun;Park, Kyung-Hwa;Kim, Yong-Hae;Yoon, Eui-Soo;Kang, Tae-Jin;Park, Kwang-Tae
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 1999.10a
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    • pp.46-57
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    • 1999
  • Ginseng(Panax ginseng C.A. Meyer) is important medicinal plant but requires 4-year cultivation for root harvest because of slow growth. In contrast, ginseng callus and hairy roots grow vigorously and may Produce the same or more biologically active compounds for human health than natural ginseng roots. Therefore, ginseng callus and hairy roots can be used for commercial purposes. Polyacetylene, one of anti-cancer compounds in ginseng, was not detected in the callus cultured on the medium containing 2, 4-B, but cells derived from the callus growth was excellent, The ginseng calli cultured on the medium containing 2mg11 CPA and 0.05mg/1 BA was grown vigorously and produced panaxydol, one of ginseng polyacetylene. The biosynthesis of polyacetylene in callus was not affected by addition of NAA and sucrose in media. The SH medium was better than the MS medium for ginseng callus growth and biosynthesis of panaxydol. Another ginseng anti-cancer compounds, ginsenoside-Rg$_3$, Rh$_1$and Rh$_2$ were detected in ginseng hairy roots by heat treatment. Those of Panax ginseng were obtained after root disks of three-year old roots were infected with Agrobacterium rhizogenes Rl000 $A_4$T in dark condition after one month of culture. The optimum growth of hairy roots was achieved in the culture of 1/2 MS liquid medium in dark(22$^{\circ}C$) under 60 rpm gyratory shaking. Hairy roots grew well in 5 ι Erlenmeyer flasks, 1ι roller drums, 10ι jar-fermenters, and especially in 20ι air-lift .culture vessels. All heat treatments had remarkably different ginsenoside contents. Eleven ginsenosides were determined in heat treatment, eight in freeze dried hairy roots. Contents of ginsenoside-Rbl , Rb2, Rc, Rd. Re, Rf, and Rg$_1$tested in all heat treatments were less than those of freeze dried hairy roots. Contents of glnsenoside-Rg$_2$ in heat treatment for 1 hour at 105$^{\circ}C$ was 4.92mg/g dry wt, 3.9 times higher than 1.27 mg/g dry wt of freeze dried hairy roots. The optimum condition of heat treatment for the production of ginsenoside-Rg$_3$and Rhl was 2 hours at 105$^{\circ}C$, and ginsenoside content was 2.58mg/g dry wt and 3.62mg/g dry wt, respectively. The production of ginsenoside-Rh2 was the highest in heat treatment for 2 hours at 105$^{\circ}C$ among treatments examined, and ginsenoside-Rh$_2$content was 1.08mg/g dry wt.

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In vitro seed germination and callus formation on flower bud of Korean mistletoe ( Viscum album L. var. cololatum [Kom.] Ohwi) (겨우살이 종자 발아 및 화아 배양에 의한 캘러스 형성)

  • Kim, Suk-Weon;Ko, Suk-Min;Liu, Jang-R.
    • Journal of Plant Biotechnology
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    • v.35 no.1
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    • pp.47-53
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    • 2008
  • Effects of growth regulators and culture conditions on seed germination, haustorium development, and callus formation of Korean mistletoe (Viscum album var. coloratum (Kom.) Ohwi) were described. Histological examination showed that seed of V. album contained one or two zygotic embryos with rod shape, and actively dividing cells were mainly distributed in radicle region rather than cotyledon of zygotic embryo. The most significant factor for seed germination and haustorium development of V. album was the requirement of the light. Various growth regulators examined in this study failed to substitute the effect of the light on seed germination. The frequency of callus formation was highest at 27.3% when flower buds were cultured onto B5 medium containing $0.1\;mgl^{-1}$ IAA. Explants from other organs were recalcitrant in forming calluses. Culture conditions described in this study could be applied for production of useful metabolites and multiplication of V. album in future.

Plant Regeneration from Internode Explants of Actinidia arguta and its Histological Observation (다래(Actinidia arguta)의 절간절편체로부터 식물체 재분화 및 해부학적 관찰)

  • 김회경;김준철;진창덕
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.5
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    • pp.263-268
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    • 1997
  • Effects of plant growth regulators and condition on shoot induction from internode explants of Actinidia arguta were investigated. Optimal condition for shoot induction was obtained when internode explants were cultured on MS medium containing 3.0 mg/L zeatin. The frequency showed about 62.5% under darks condition at 27$^{\circ}C$ after 6 weeks of culture. Regenerated shoots were rooted on WPM medium supplemented with 0.5 mg/L NAA and the regenerants were acclimated to an artificial soil with mixture (perlite : vermiculite = 1:1, v/v) for further growth and successfully transplanted to the soil in a highly humidified greenhouse. Histological observations revealed that meristemoids were formed from small and isodiametric cells, and then developed to shoot primordia.

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