• Title/Summary/Keyword: Co-incubation

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Genetic characteristics of the novel insect pathogenic Bacillus thuringiensis subsp. aizawai strain (새로운 곤충병원성 Bacillus thuringiensis subsp. aizawai 균주의 유전학적 특성)

  • Seo, Mi Ja;Youn, Young Nam;Yu, Yong Man;Kim, Ki Su
    • Korean Journal of Agricultural Science
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    • v.41 no.4
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    • pp.351-359
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    • 2014
  • For identifying the plasmid DNA coding cry gene of Bacillus thuringiensis subsp. aizawai KB098 with high insecticidal activity against Spodoptera exigua, mutant isolates with no crystal protein were produced by $42^{\circ}C$ incubation condition and then mutant plasmid DNA band patterns were compared with those of KB098. KB098 isolates had 4 cry genes, cry1Aa, cry1Ab, cry1C, cry1D, and also had been found seven plasmid DNA. Though the SDS-PAGE experiment, it was confirmed that mutant didn't produce 130~145kDa protein band involved in bipyramidal shape crystal. Also, five mutant isolates had no cry genes coding plasmid DNA in PCR. In result of comparison the plasmid DNA of KB098 and 5 mutant isolates, only 1 plasmid DNA band was left out in mutant plasmid DNA pattern, so that the missing band was extracted from the gel. The missing(disappeared) plasmid DNA was the largest molecular size among the 7 plasmid DNA of KB098 and it was also confirmed this plasmid DNA had all 4 cry genes through PCR.

Nitrogen mineralization of oil cakes according to changes in temperature, moisture, soil depth and soil texture (유박의 온도, 수분, 토심, 및 토성에 따른 질소의 무기화)

  • Cho, Sung-Hyun;Chang, Ki-Woon
    • Journal of the Korea Organic Resources Recycling Association
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    • v.15 no.1
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    • pp.149-158
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    • 2007
  • To investigate the characteristics of the nitrogen mineralization of oil cakes according to changes in temperature(10, 20, $30^{\circ}C$) moisture(40, 50, 60, 70% of field capacity in loam and 50, 60, 70, 80% of field capacity in sandy loam), mineral nitrogen was measured in soil after incubation for 30 days. In addition, the mineralization in sandy loam and loam which had different soil texture were compared. According to incubating the castor seed, soybean, and rice bran cakes with soil, the higher the temperature and moisture content were, the higher the content of mineral nitrogen was observed. The content of mineral nitrogen was higher in sandy loam than loam. The content of mineral nitrogen was decreased with soil depth increasing and was also higher in sandy loam than loam.

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Xylanase Production by Mixed Culture Using Crude Hemicellulose from Rice Straw Black Liquor and Peat Moss as an Inert Support

  • Shata, Hoda Mohamed Abdel Halim;El-Deen, Azza Mohmed Noor;Nawwar, Galal Abdel Moen;Farid, Mohmed Abdel Fattah
    • Journal of Applied Biological Chemistry
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    • v.57 no.4
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    • pp.313-320
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    • 2014
  • Black liquor (BL) is a by-product of rice straw pulping process. It is a low costs raw material for production value-adding proteins and enzymes, which has been paid more and more attention to reduce its environmental pollution. Mixed cultures of micelial fungi, Trichoderma reesei Northern Regional Research Laboratory (NRRL)11236, Trichoderma reesei NRRL 6165 and Aspergillus niger strains NRC 5A, NRC 7A, and NRC 9A were evaluated for their ability to produce xylanase using crude hemicellulose (CHC) prepared from BL and peat moss as an inert support under solid state fermentation (SSF). The most potent strains, A. niger NRC 9A (818.26 U/g CHC) and T. reesei NRRL 6165 ($100.9{\pm}57.14$ U/g CHC), were used in a mixed culture to enhance xylanase production by co-culturing under SSF. In the mixed culture, xylanase production ($1070.52{\pm}12.57$ U/g CHC) was nearly1.3 and 10.6-fold increases over the activities attained in their monocultures, A. niger NRC 9A and T. reesei NRRL 6165, respectively. Optimization of the culture parameters of the mixed culture SSF process, concentration of ammonium sulfate and corn steep liquor, CHC/peat moss ratio, inoculum size and ratios of the two strains, initial pH value, initial moisture content and incubation time, exhibited a significant increase ($2414.98{\pm}84.02$ U/g CHC) in xylanase production than before optimization.

Effect of Adenosine on the Release of $[^3H]-5-hydroxytryptamine$ during Glucose/Oxygen Deprivation from Rat Hippocampal Slices (흰쥐 해마절편에서 포도당/산소 고갈에 의한 5-hydroxytryptamine 유리변동에 미치는 Adenosine의 영향)

  • Cha, Kwang-Eun;Pae, Young-Sook;Lee, Kyung-Eun
    • The Korean Journal of Physiology and Pharmacology
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    • v.1 no.6
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    • pp.657-664
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    • 1997
  • The effects of adenosine, adenosine A1 receptor antagonist (DPCPX), or NMDA receptor antagonist (APV) on the spontaneous release of $[^3H]-5-hydroxytryptamine$ ($[^3H]-5-HT$) during normoxic/normoglycemic or hypoxic/hypoglycemic period were studied in the rat hippocampal slices. The hippocampus was obtained from the rat brain and sliced $400\;{\mu}m$ thickness with the tissue slicer. After 30 min's preincubation in the normal buffer, the slices were incubated for 30 min in a buffer containing $[^3H]-5-HT$ ($0.1\;{\mu}M,\;74{\mu}Ci/8\;ml$) for uptake, and washed. To measure the release of $[^3H]-5-HT$ into the buffer, the incubation medium was drained off and refilled every ten minutes through sequence of 14 tubes. Induction of glucose/oxygen deprivation (GOD; medium depleting glucose and gassed with 95% $N_2/5%\;CO_2$) was done in 6th and 7th tube. The radioactivities in each buffer and the tissue were counted using liquid scintillation counter and the results were expressed as a percentage of the total radioactivities. When slices were exposed to GOD for 20 mins, the spontaneous release of $[^3H]-5-HT$ was markedly increased and this increase of $[^3H]-5-HT$ release was blocked by adenosine ($10\;{\mu}M$) or DL-2-amino-5-phosphonovaleric acid (APV; $30\;{\mu}M$). Adenosine $A_1$ receptor specific antagonist, 8-cyclopentyl-1,3-dipropylxanthine (DPCPX) exacerbate GOD-induced increase of spontaneous release of $[^3H]-5-HT$. These results suggest that Adenosine may play a role in the GOD-induced spontaneous release of $[^3H]-5-HT$ through adenosine $A_1$ receptor activity.

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Influence of Some Flavonoids on N-Nitrosoproline Formation in vitro and in vivo (시험관 및 생체조건하에서 니트로소프로린 생성에 미치는 수종 플라보노이드 화합물의 영향)

  • Lee, Ji-Hyeon;Park, Jae-Sue
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.22 no.3
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    • pp.266-272
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    • 1993
  • Some compounds including flavonoids were tested as scavenger of nitrite which is believed to participate in the formation of N-nitroso compound. Many were found to be potent scavengers and the most potent ones were ascorbic arid, potassium thiocyanate, chlorogenic acid, catechin, morin, luteolin, luteolin-7-glucoside, and naringenin. To evaluate the influence of the above compounds on the endogenous formation of N-nitroso compound, the amount of nitrosoproline (NPRO) was examined by co-incubation of nitrite, proline, and test compounds at various concentrations. The analysis of NPRO by High Performance Liquid Chromatography (HPLC) was newly developed. Most compounds except ascorbic acid and chlorogenic acid were found to be no effects or activatory effects on NPRO formation. From the results obtained, it was suggested that most flavonoids which are contained in our customary diets were not associated with the inhibition of NPRO formation.

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A Rat Liver Lysosomal Membrane Flavin-Adenine Dinucleotide Phosphohydrolase

  • Shin, Hae-Ja;Lim, Woon-Ki
    • BMB Reports
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    • v.29 no.3
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    • pp.253-260
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    • 1996
  • An enzyme that hydrolyzes flavin-adenine dinucleotide (FAD) was found to be present in rat liver lysosomal membrane prepared from Triton WR-1339 filled lysosomes (tritosomes) purified by flotation on sucrose. This FAD phosphohydrolase (FADase) exhibited optimal activity at pH 8.5 and had an apparent Km of approximately 3.3 mM. The activity was decreased 50~70% by dialysis against EDTA and this was restored by $Zn^{2+}$, $Mg^{+2}$, $Hg^{+2}$, and $Ca^{+2}$ ions inhibited the enzyme, but $F^-$ and molybdate had no effect. The enzyme was also inhibited by p-chloromercuribenzoate (pCMB), reduced glutathione and other thiols, cyanide, and ascorbate. The presence of ATP, ADP, AMP. ${\alpha}-{\beta}-methylene$ ATP, AMP-p-nitrophenyl phosphate (PNP), GMP, and coenzyme A (CoA) decreased the activity on FAD, but pyrimidine nucleotides, adenosine, adenine, or $NAD^+$ were without effect. Phosphate stimulated the activity slightly. FAD phosphohydrolase activity was separated from ATPase and inorganic pyrophosphatase activities by solubilization with detergents and polyacrylamide gel electrophoresis and by linear sucrose density gradient centrifugation suggesting that the enzyme is different from ATPase, inorganic pyrophosphatase, and soluble lysosomal FAD pyrophosphatase. Paper chromatography showed that FAD was hydrolyzed to flavin mononucleotide (FMN) and AMP which were further hydrolyzed to riboflavin and AMP by phosphatases known to be present in lysosomal membranes. Incubation of the intact Iysosomes with pronase showed that the active site of FAD phosphohydrolase must be oriented to the cytosol. The FAD hydrolyzing activity was detected in Golgi, microsome, and plasma membrane, but not in mitochondria or soluble lysosomal preparations.

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Inhibition of L-DOPA-Induced Increase in Dopamine Content by $(1R,9S)-{\beta}-Hydrastine$ Hydrochloride in PC12 cells

  • Yin, Shou-Yu;Lee, Jae-Joon;Kim, Yu-Mi;Jin, Chun-Mei;Yang, Yoo-Jung;Kang, Min-Hee;Lee, Myung-Koo
    • Natural Product Sciences
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    • v.10 no.3
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    • pp.119-123
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    • 2004
  • The effects of BHSH on L-DOPA-induced increase in dopamine content in PC12 cells were investigated. L-DOPA treatment at $20\;or\;50\;{\mu}M$ increased dopamine content after both 24 and 48 h of incubation in PC12 cells. However, the co-treatments of BHSH $(10-50\;{\mu}M)$ with L-DOPA $(20\;or\;50\;{\mu}M)$ significantly inhibited the increase of dopamine content induced by L-DOPA. BHSH treatment at $10-50\;{\mu}M$ significantly inhibited basal aromatic L-amino acid decarboxylase (AADC) activity in a concentration-dependent manner at 15 min, and then AADC activity was rapidly recovered to the control level at about 2 h. These results indicate that the inhibition of AADC activity by BHSH was, in part, contributed to the early-stage decrease of dopamine content induced by LDOPA in PC12 cells. Taken together, it is proposed that the short-term inhibition of dopamine biosynthesis by BHSH was mediated by the regulation of tyrosine hydroxylace (TH).

Impact of Physical, Chemical and Biological Factors on Lily (Lilium longiflorum cv. Georgia) Pollen Growth and GUS Expression Via Agro-infiltration (물리적, 화학적, 생물적 요인에 의한 백합 (Lilium longiflorum cv. Georgia) 화분의 생장 및 Agro-Infiltration을 이용한 GUS 발현)

  • Park, Hee-Sung
    • Journal of Plant Biotechnology
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    • v.31 no.4
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    • pp.279-283
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    • 2004
  • To lily (Lilium longflorum cv. Georgia) pollen, impacts by some physical, chemical and biological factors were examined in respects of its growth and transient gene expression via agro-infiltration. Rolling movement in liquid medium or vacuum pressure during Agro-infiltration was regarded as a impact that should be minimized for normal pollen growth. Pollen growth was maintained well in relatively broad range of temperature (19 to 27$^{\circ}C$) or pH (5.0 to 8.0). Chemical factors such as cefotaxime (up to 300mg/L), acetosyringone (up to 800 $\mu$M) and syringealdehyde (up to 800 $\mu$M) did not show any harmful effects but kanamycin severely did even at concentration as low as 25mg/L in some cases. For GUS gene expression, acetosyringone at 200 to 400 $\mu$M slightly improved the efficiency while syringealdehyde did not. Brief agro-infiltration followed by 18 hr of co-incubation of pollen along with Agrobacterium was suggested as a condition basically required for the transient expression system using lily pollen regardless of the presence of acetosyringone.

High Postnatal Survival and Efficacy of Female-Derived Donor Cells in the Productive of Somatic Cloned Piglets

  • Cho, Seong-Keun;Park, Mi-Ryung;Hwang, Kyu-Chan;Kwon, Deug-Nam;Im, Yeo-Jeoung;Park, Ju-Joung;Son, Woo-Jin;Kim, Jin-Hoi
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.33-33
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    • 2003
  • This study was conduct to compare the efficacy to produce male and female somatic cloned piglets. Maturation of porcine COCs was accomplished by incubation in NCSU-23 medium supplemented with 0.6 mM cysteine, 10% porcine follicular fluid, 1mM dibutyryl cyclic adenosine monophosphate (dbc-AMP, Sigma, USA), and 0.1 IU/ml human menopausal gonadotrophin (hMG, Teikokuzoki, Japan) for 20h and then cultured without dbcAMP and hMG for another 18 to 24 h. Female and male fetal cells were isolated from each fetus, cultured in ES-DMEM medium containing 10% FCS. Enucleated oocytes were fused with fetal fibroblasts (passage 4 to 15). Reconstructed embryos were cultured in NCSU-23 with 4 mg/ml BSA under mineral oil at 39$^{\circ}C$ in 5% $CO_2$ in air. A total of 12,328 nuclear-transferred embryos (1- to 4-cell stage) were surgically transferred into 69 surrogate gilts. Three recipients aborted during the period of conception. Three gilts delivered eleven female piglets, and five recipients gave rise to birth 22 male piglets. The average birth weigh of the cloned piglets was 1.52 kg (1.38~1.83 kg) in female piglets and 0.84 kg (0.45~1.25 kg) in male piglets. Alive cloned pigs was seven in female piglets (63.6%) and four in male piglets (18.2%). The other two recipients is ongoing. This study suggests that female-derived fetal cell as a nuclear donor has more capability on production of cloned piglets than male.

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Production of Reuterin by Batch and Continuous Reactor and Antimicrobial Characteristics of Reuterin (회분식과 연속식에 의한 루테린 생산 및 루테린의 항균 특성)

  • Yum, Eun-Mi;Kim, Ji-Yeun;Shin, Hyun-Kyung;Ji, Geun-Eog
    • Korean Journal of Food Science and Technology
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    • v.36 no.1
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    • pp.111-115
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    • 2004
  • Reuterin production efficiency of Lactobacillus reuteri, which converts glycerol into reuterin (antimicrobial substance) under anaerobic condition, was examined. When compared at $32,\;37,\;and\;42^{\circ}C$, production rate and total amount produced increased with increasing incubation temperature. Reuterin production terminated earlier at $42^{\circ}C$ than at $32\;and\;37^{\circ}C$. Presence of various amino acids in the reaction mixture generally decreased reuterin production, whereas proline did not inhibit reuterin production. A continuous-type reactor in which glycerol was passed through the chamber containing L. reuteri cells produced greater amount of reuterin than when batch-type process was used.