• Title/Summary/Keyword: co-cultures

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Effect of Culture Conditions on Survival of Frozen-Thawed Blastocysts Fertilized In Vitro (소 체외수정란의 배양조건이 동결-융해 배반포의 생존에 미치는 영향)

  • 윤종택;이호준;노상호;정연길;박용습;최은주;이종완;김용엽;정혜영
    • Journal of Embryo Transfer
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    • v.14 no.3
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    • pp.163-169
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    • 1999
  • This study was carried out to investigate the effect of co-culture system(bovine oviduct epithelial cells; BOEC) and defined culture system(modified TALP ; mTALP) on the development of IVM-IVF embryos, and survival of in vitro produced blastocysts after freezing and thawing. Occytes from the slaugheterhous ovaries were matured and fertilized using general protocol. The results obtained were as the following: 1. Survival rates of frozen-thawed blastocysts using 10% glycerol as cryoprotectant was higher in day 7 blastocysts than in Day 8 and 9 blastocysts from co-cultrue system, but survival rate of frozen-thawed blastocysts was higher in Day 10 blastocysts than in day 8 and 9 blastocysts from defined culture system. Regardless of their age, survival rate of frozen-thawed blastocysts was significantly higher (p<0.05) in co-culture system than in defined culture system. 2. The cell number of blastocysts was significanlty higher (p<0.05) in Day 7 blasotcysts than in Day 8 and 9 blastocysts from co-cultures, but the cell number of blsstocysts was significantly higher (p<0.05) in Day 10 blastocysts than in Day 8 and 9 blastocysts from defined culture system. Regardless of the culture system, blastocysts with higher cell number showed higher survival rates after freezing and thawing.

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Synthesis of Poly[3-hydroxybutyrate-co-3-hydroxyvalerate] by Recombinant Escherichia coli from Whey (재조합 대장균에 의한 유청으로부터 Poly[3-hydroxybutyrate-co-3-hydroxyvalerate] 합성)

  • 김범수;이상엽
    • KSBB Journal
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    • v.18 no.5
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    • pp.404-407
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    • 2003
  • Two recombinant Escherichia coli strains, GCSC6576 harboring a plasmid pSYL107 containing the Ralstonia eutropha polyhydroxyalkanoate (PHA) biosynthesis genes and a fadR atoC mutant LS5218 harboring a plasmid pJC4 containing the Alcaligenes latus PHA biosynthesis genes were compared for their ability to synthesize poly(3-hydroxybutyrate-co-3-hydroxyvalerate) [P(3HB-co-3HV) from whey. The 3HV fraction could be increased by acetic acid induction and oleic acid supplementation in flask cultures of recombinant E. coli GCSC6576. With the pH-stat fed-batch culture of recombinant E. coli LS5218, we obtained a cell concentration, a P(3HB-co-3HV) concentration, a P(3HB-co-3HV) content, and a 3HV fraction of 31.8 g/L, 10.6 g/L, 33.4%, and 6.26 mol%, respectively in 39 h.

Bipolar Heterothallism, a Principal Mating System of Cordyceps militaris In Vitro

  • Shrestha Bhushan;Kim Ho Kyung;Sung Gi Ho;Spatafora Joseph W.;Sung Jae Mo
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.9 no.6
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    • pp.440-446
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    • 2004
  • Interest in in vitro study of entomopathogenic fungi, including Cordyceps species, has been increasing due to their valuable bioactive compounds and biocontrol effects. Among Cordyceps species, in vitro stromata of C militaris has been successfully produced and cultivated for industrial purposes. However, genetic study on in vitro stromata formation of C militaris has not been carried out yet. Here, relationship between mating system and perithecial stromata formation of C militaris is reported. Mating system was determined by observing perithecial stromata formation from mono-ascospore cultures and their pair-wise combinations. Certain combinations of mono-ascospore strains produced perithecial club-shaped stromata, whereas other combinations produced either no stromata or only abnormal non-perithecial stromata. Similarly, mono­ascospore cultures without combination produced either no stromata or only abnormal non­perithecial stromata. Despite obvious heterothallism, self-fertility was occasionally observed in few strains of C militaris. These observations indicated that C militaris behaves as a bipolar het­erothallic fungus and requires two mating compatible strains in order to produce regular club­shaped perithecial stromata, a fundamental requirement for its industrial cultivation.

Production of Tropane Alkaloids by Optimal Culture Conditions in Adventitious Root of Hyoscyamus niger L.

  • Park, Dong-Jin;Min, Ji-Yun;Kim, Yong-Duck;Kang, Seung-Mi;Jung, Ha-Na;Kang, Hoon-Serg;Choi, Myung-Suk
    • Korean Journal of Medicinal Crop Science
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    • v.13 no.5
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    • pp.237-241
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    • 2005
  • Scopolamine and hyoscyamine are important anticholinergic compounds obtained from Hyoscyamus niger. Adventitious roots induced from rhizome of H. niger and roots were cultured in SH medium supplemented with 3% (w/v) sucrose and 0.5 mg/L IBA. Roots were grown rapidly after 10 days of cultures. Scopolamine production was increased 7 times and hyoscyamine production was increased 12 times after 10 day of cultures. SH medium was best in root growth. But, highest scopolamine productivity was observed in WPM medium, followed White medium and best hyoscyamine productivity was resulted in MS medium. Sucrose was increased scopolamine and hyoscyamine production were increased the medium supplemented by sucrose comparing to than those by other carbon sources.

Effect of Polyamines on Indigo Biosynthesis in Hairy Root Cultures of Polygonum tinctorium Lour. (Polyamine이 쪽 모상근배양에서 인디고 생합성에 미치는 영향)

  • Kim, Jin-Nam;Jang, Hong-Gi;Park, Sang-Un;Ryu, Hwa-Won
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.51 no.spc1
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    • pp.247-250
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    • 2006
  • We herein studied the effect of polyamines (putrescine, spermidine and spermine) on growth and indigo biosynthesis in hairy root cultures of Polygonum tinctorium Lour. Our results revealed that polyamine treatment increased hairy root growth and indigo biosynthesisat all tested concentrations, with the highest growth rate (4.4 g/ flask) and indigo yield (216 ug/g) induced by 70 mg/L putrescine. These results show far the first time that the growth rates and indigo biosynthesis of Polygonum tinctorium hairy roots may be improved by addition of polyamines to the liquid culture medium.

Nitrogen Removal from Wastewaters by Microalgae Without Consuming Organic Carbon Sources

  • Lee, Kwang-Yong;Lee, Choul-Gyun
    • Journal of Microbiology and Biotechnology
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    • v.12 no.6
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    • pp.979-985
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    • 2002
  • The possibility of microalgal nitrogen treatment was tested in wastewaters with a low carbon/nitrogen (C/N) ratio. Chlorella kessleri was cultured in the two different artificial wastewaters with nitrate as a nitrogen source: one contained glucose for an organic carbon source and the other without organic carbon sources. The growth rates of the two cultures were almost identical when the aeration rate was over 1 vvm. These results suggest that microalgae could successfully remove nitrogen from wastewater, as far as the mass transfer of $CO_2$, was not limited. Nitrate was successfully reduced to below 2 mg $NO_3^-$-N/ml from the initial nitrate concentration of 140 mg $NO_3^-$-N/ml in 10 days, even in the wastewater with no organic carbon source. Similar results were obtained when ammonium was used as the sole nitrogen source instead of nitrate. Higher concentrations of nitrogen of 140, 280, 560 and 1,400 mg/ml were also tested and similar amounts of nitrogen were removed by algal cultures without showing any substrate inhibition.

Production of 8-epi-Tomentosin by Plant Cell Culture of Xanthium strumarium

  • Park, Jae-Sung;Yi, Gi-Hwan;Nam, Min-Hee;Park, Sun-Ho
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.6 no.1
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    • pp.51-55
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    • 2001
  • This study was conducted to establish a plant cell culture system for the production of medically important secondary metabolites from Xanthium strumarium. The effects of plant growth regulators including NAA, 2,4-D, kinetin, and ABA were examined in terms of callus induction, maintenance of callus and suspension cultures. It was shown that callus was induced upon treatment with NAA while embryo was induced after treatment with 2,4-D. Callus formation was further improved by treatment with ABA and NAA. The level of callusing increased by 17-29% for the seed case, cotyledon, leaf, and hypocotyl and by 96% in the case of the root. Suspension cell lines were established using calli produced from cotyledon, hypocotyl and root and cultured at 25$\^{C}$ under light conditions. The cells grew up to 15g/L with NAA 2ppm, BA 2ppm, and ABA 1ppm treatment. Supernatants of suspension cultures of cell lines derived from coyledon and hypocotyl produced some distinctive secondary metabolites, one of which was identified as 8-epi-tomentosin, which belongs to the xanthanolides. The amounts of 8-epi-tomentosin produced by the cotyledon- and hypocotylderived cell lines were 13.4mg/L and 11.0mg/L, respectively.

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Removal of Residual Toxic Phytohormone in Suspension Cultures of Panax vietnamensis (베트남 인삼세포 현탁 배양에서 잔류 독성 식물호르몬 제거)

  • Lee, Seung-Ho;Kim, Nam-Hyuck;Kim, Ki-Hyun;Quang, Le Bach;Luong, Hoang Van;Byun, Sang-Yo
    • KSBB Journal
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    • v.23 no.2
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    • pp.183-185
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    • 2008
  • Studies were made to remove the toxic phytohormone, 2,4-D, in suspension cultures of Panax vietnamensis. Cells grown in normal MS medium with 2,4-D were inoculated and grown in the MS medium without hormone. Not a big difference was observed in growth characteristics between media with and without 2,4-D. The 2,4-D in the culture, however, was completely removed. During the culture, the residual 2,4-D was consumed rapidly at the early growth stage. The intra-cellular 2,4-D was consumed first and the 2,4-D in the medium was used afterward.

Regulation of γ-Aminobutyric Acid Production in Tobacco Plants by Expressing a Mutant Calmodulin Gene

  • Oh, Suk-Heung;Cha, Youn-Soo
    • Journal of Applied Biological Chemistry
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    • v.43 no.2
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    • pp.69-73
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    • 2000
  • In order to understand the biological role of calmodulin in plants, transgenic plants expressing a mutant calmodulin (VU-4, Iys to ile-115) have been analyzed. We found that tobacco plants expressing VU-4 calmodulin have approximately twofold higher $\gamma$-aminobutyric acid (GABA) levels than the control plants. Cell suspension cultures established from the stem explants of the transgenic tobacco seedlings also have higher levels of GABA than the control cell cultures. Specific activity of glutamate decarboxylase (GAD), which catalyzes the decarboxylation of glutamate to $CO_2$ and GABA, of the transgenic tobacco cell extracts was about twofold higher than the activity of the control cell extracts. Western-blot analysis showed that the GAD is highly expressed in the transgenic tobacco plants. GAD partially purified from tobacco cell extracts showed approximately threefold $Ca^{2+}$/calmodulin-dependent activation. These data suggest that GABA synthesis in the transgenic tobacco plants is elevated, possibly due to higher levels of the calmodulin-dependent GAD enzyme and/or as a result of enhanced activation due to increased levels of the foreign calmodulin.

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Physiological Characteristics of Immobilized Streptomyces Cells in Continuous Cultures at Different Dilution Rates

  • Kim, Chang-Joon;Chang, Yong-Keun;Chun, Gie-Taek;Jeong, Yeon-Ho;Lee, Sang-Jong
    • Journal of Microbiology and Biotechnology
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    • v.12 no.4
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    • pp.557-562
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    • 2002
  • Physiological characteristics such as specific productivity, morphology of Streptomyces cells Immobilized on celite beads, and operational stability at different dilution rates were investigated in continuous immobilized-cell cultures for the production of kasugamycin. At a dilution rate (D) of 0.05 $h^{-1}$, a relatively high specific productivity was attained and the loss of cell-loaded beads was negligible. At D=0.1 $h^{-1}$, a higher specific productivity and cell concentration could be obtained, resulting in a significantly improved volumetric kasugamycin productivity. However, no stable operation could be maintained due to a significant loss of cell-loaded beads from the reactor that was caused by their fluffy morphology developed in the later stage. At D=0.2 $h^{-1}$, the production of kasugamycin and cell growth were observed to be severely inhibited by the high concentration of residual maltose.