• Title/Summary/Keyword: polyethylene glycol(PEG)

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Development of Yeast-Vector System for Eukaryotic Gene Cloning - Optimum Condition for Intact Yeast Cell Transformation and Plasmid Stability in the Transformants - (진핵생물 유전자 조작을 위한 효모 vector계 이용에 관한 기초연구 -생효모 형질전환 최적조건과 숙주별 plasmid안정성에 관하여 -)

  • 기우경;조성환;김범규;조무제
    • Microbiology and Biotechnology Letters
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    • v.14 no.2
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    • pp.125-131
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    • 1986
  • In order to obtain the optimum conditions for intact yeast cell transformation in the various yeast host-vector systems, 3 yeast plasmid vectors, YRp7, YEpl3 and YIp5 were introduced into 5 yeast hosts, Saccaromyces cervisiae Dl3-1A, DKD-5D, DBY-746, MC-16 and S2022D with various transformation conditions, and plasmid stabilities in all the transformants were also observed. The highest transformation frequencies in all the host-vector system were obtained in the 16 hour Cultured cell (5.4 $\times$ 10$^6$ - 2.4 $\times$ 10$^8$cells/$m{\ell}$) treated with 0.1-0.2 M lithium chloride in 0.1 M tris-HCl (pH 7.6), 35% polyethylene glycol 4000, and heat-shocked at 42$^{\circ}C$ for 5 minutes after 60 minutes of induction. The intact cell transformation got more transformation frequency in DKD-5D (YRp7) and DBY-746 (YEpl3) than protoplast transformation, but reverse tendency was observed in DKD-5D (YEp13) and Dl3-lA (YRp7). The transformants, D13-1A (YRp7) and DKD-5D (YRp7) were very unstable in selective medium, with 80 to 85% of the transformants losing the plasmid after 70 generations, but the transformants, DKD-5D (YEpl3) and DBY-746 (YEpl3) were quite stable, with 35% of the transformants losing the plasmid.

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Purification of Inositol Triphosphate Kinase from Bovine Brain (소의 뇌로부터 Inositol Triphosphate Kinase의 정제)

  • Kim, Jung-Hye;Lee, Jae-Tae
    • Journal of Yeungnam Medical Science
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    • v.13 no.1
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    • pp.46-58
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    • 1996
  • Inositol 1,4,5-triphosphate($InsP_3$) is a second messenger for mobilizing intracellular $Ca^{2+}$. It can be dephosphorylated by soluble and particulate forms on $InsP_3$ 5-phosphatase, or phosphorylated to produce inositol 1,3,4,5-tetrakisphosphate($InsP_3$) by $InsP_3$ 3-kinase. These enzymes represent possible targets for the regulation of the $InsP_3/InsP_4$ signal. $InsP_3$ 3-kinase which catalyses th ATP-dependent phosphorylation of $InsP_3$ was purified from bovine brain tissue. All operation were carried out at $4^{\circ}C$. Fresh tissure was homogenized and centrifuged. The supernatant was pooled. Proteins were precipitated from 10% polyethylene glycol, and suspended solution was applied to DEAE cellulose column for chromatography. As the result of above procedure, two isozymes of $InsP_3$ 3-kinase, I and II were obtained. Each isozyme was applied to Matriz green gel, Calmodulin-Affigel 15 column and subsequent phenyl-TSK HPLC column. Specific activites(SA) and fold of puriety were observed at each purification step of chromatography. At DEAE cellulose chromatography, SA were I, 0.6 and II, 4.8 nM/min/mg, and folds were I, 17.2 and II, 16.6. At Matrix green gel chromatography, SA were I, 18 and II, 11 nM/min/mg, folds were I, 62.1 and II, 38.0. At calmodulin-Affigel 15 column chromatography, SA were I, 19 and II, 13 nM/min/mg, folds were I, 65.5 and II, 44.8. Finally $InsP_3$ kinase I and II were purified 3,103-fold and 2,310-fold, and SA were I, 900 and II, 670 nM/min/mg, respectively. SDS-polyacrylamide gel electrophoresis elucidated 3 distinct fractions of Mr of 145,000, 85,000 and 69,500 from isozyme I, and 2 distinct fractions of Mr of 79,000 and 57,000 from isozyme II.

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Assessment of Discoidal Polymeric Nanoconstructs as a Drug Carrier (약물 운반체로서의 폴리머 디스크 나노 입자에 대한 평가)

  • BAE, J.Y.;OH, E.S.;AHN, H.J.;KEY, Jaehong
    • Journal of Biomedical Engineering Research
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    • v.38 no.1
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    • pp.43-48
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    • 2017
  • Chemotherapy, radiation therapy, and surgery are major methods to treat cancer. However, current cancer treatments report severe side effects and high recurrences. Recent studies about engineering nanoparticles as a drug carrier suggest possibilities in terms of specific targeting and spatiotemporal release of drugs. While many nanoparticles demonstrate lower toxicity and better targeting results than free drugs, they still need to improve their performance dramatically in terms of targeting accuracy, immune responses, and non-specific accumulation at organs. One possible way to overcome the challenges is to make precisely controlled nanoparticles with respect to size, shape, surface properties, and mechanical stiffness. Here, we demonstrate $500{\times}200nm$ discoidal polymeric nanoconstructs (DPNs) as a drug delivery carrier. DPNs were prepared by using a top-down fabrication method that we previously reported to control shape as well as size. Moreover, DPNs have multiple payloads, poly lactic-co-glycolic acid (PLGA), polyethylene glycol (PEG), lipid-Rhodamine B dye (RhB) and Salinomycin. In this study, we demonstrated a potential of DPNs as a drug carrier to treat cancer.

Development of Composite Hollow Fiber Membranes for Olefin Off-gas Recovery (올레핀 배가스의 분리를 위한 중공사형 복합막의 개발)

  • Kim Jeong-Hoon;Choi Seung-Hak;Lee Soo-Bok
    • Membrane Journal
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    • v.15 no.2
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    • pp.157-164
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    • 2005
  • In this study, composite hollow fiber membranes were developed for the recovery of olefin monomers in polyolefin industry off-gases. Polyetherimide (PEI) hollow fiber support membranes were fabricated from spinning solutions containing PEI, NMP and polyethylene glycol (PEG). The influence of dope solution and inner coagulant composition on the permeation properties and structure of hollow fiber supports was examined. PDMS was used as a selective layer and coated on PEI hollow fiber support. The thickness of active layer was controlled by changing coating solution concentration. The permeation properties of hollow fiber supports and composite membranes were characterized with a pure gas permeation test. The optimized composite hollow fiber membrane has $10\;{\mu}m$ selective layer and shows excellent separation performance; the ideal selectivity of olefins over nitrogen is in the following order: 1-butylene (6.4) > propylene (17) > ethylene (97), which selectivity data are similar to the intrinsic olefin/nitrogen selectivities of PDMS. This confirms that the new composite hollow fiber membranes suitable for olefin off-gas recovery has developed successfully.

Transcriptome Profiling and Characterization of Drought-Tolerant Potato Plant (Solanum tuberosum L.)

  • Moon, Ki-Beom;Ahn, Dong-Joo;Park, Ji-Sun;Jung, Won Yong;Cho, Hye Sun;Kim, Hye-Ran;Jeon, Jae-Heung;Park, Youn-il;Kim, Hyun-Soon
    • Molecules and Cells
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    • v.41 no.11
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    • pp.979-992
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    • 2018
  • Potato (Solanum tuberosum L.) is the third most important food crop, and breeding drought-tolerant varieties is vital research goal. However, detailed molecular mechanisms in response to drought stress in potatoes are not well known. In this study, we developed EMS-mutagenized potatoes that showed significant tolerance to drought stress compared to the wild-type (WT) 'Desiree' cultivar. In addition, changes to transcripts as a result of drought stress in WT and drought-tolerant (DR) plants were investigated by de novo assembly using the Illumina platform. One-week-old WT and DR plants were treated with -1.8 Mpa polyethylene glycol-8000, and total RNA was prepared from plants harvested at 0, 6, 12, 24, and 48 h for subsequent RNA sequencing. In total, 61,100 transcripts and 5,118 differentially expressed genes (DEGs) displaying up- or down-regulation were identified in pairwise comparisons of WT and DR plants following drought conditions. Transcriptome profiling showed the number of DEGs with up-regulation and down-regulation at 909, 977, 1181, 1225 and 826 between WT and DR plants at 0, 6, 12, 24, and 48 h, respectively. Results of KEGG enrichment showed that the drought tolerance mechanism of the DR plant can mainly be explained by two aspects, the 'photosynthetic-antenna protein' and 'protein processing of the endoplasmic reticulum'. We also divided eight expression patterns in four pairwise comparisons of DR plants (DR0 vs DR6, DR12, DR24, DR48) under PEG treatment. Our comprehensive transcriptome data will further enhance our understanding of the mechanisms regulating drought tolerance in tetraploid potato cultivars.

Efficient Selection Method for Drought Tolerant Plants Using Osmotic Agents

  • Park, Dong-Jin;Im, Hyeon-Jeong;Jeong, Mi-Jin;Song, Hyeon-Jin;Kim, Hak-Gon;Suh, Gang-Uk;Ghimire, Balkrishna;Choi, Myung-Suk
    • Journal of Forest and Environmental Science
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    • v.34 no.3
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    • pp.224-234
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    • 2018
  • An efficient method to select drought tolerant Korean native plants using in vitro culture system was established in this study. While the plant growths and root inductions of each plant were proportionately affected by concentrations of mannitol on in vitro culturing seven plant species to test tolerance to osmotic stress, growth index (GI) and number of root induction of Chrysanthemi zawadskii var. latilobum and Dianthus chinensis var. semperflorens plantlets were higher than the others in 125mM mannitol. In test with polyethylene glycol (PEG), plantlets of C. zawadskii var. latilobum and D. chinensis var. semperflorens showed higher GI and number of root induction than the others in 33.3mM. On testing whether the well grown plants under osmotic stress are tolerant to virtual drought stress, there were significant differences in the withering rates of C. zawadskii var. latilobum and D. chinensis and those of were Aster yomena and Centaurea cyanus after 12 days without watering. It was found that significantly lower stomata numbers were shown in both drought tolerant plants than the sensitive plants. Averages of the stomata circumferences and the stomata area in the plantlets of the tolerant species were larger than those of the sensitive plants D. chinensis var. semperflorens showed the lowest transpiration level per unit area. The highest stomatal area per unit area was found in C. zawadskii, followed by D. chinensis var. semperflorens, Aster yomena and C. cyanus. In conclusion, C. zawadskii var. latilobum and D. chinensis var. semperflorens were more tolerant to drought than other two species. Furthermore in vitro selection was successfully used to screen drought tolerance species of native plant species.

Effect of Osmotic and Solid Matrix Priming to Improve Germination and Early Growth of Carrot and Onion Seed (Osmotic Priming 및 Solid Matrix Priming 처리에 의한 당근과 양파종자의 발아증진과 초기생육에 미치는 영향)

  • 강점순
    • Journal of Bio-Environment Control
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    • v.13 no.3
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    • pp.135-142
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    • 2004
  • The objective of this study was carried out to determine which method is effective for improving seed germination in carrot and onion among osmotic or solid matrix priming. Seeds were osmotic primed with -0.5 MPa polyethylene glycol 8000 (PEG) solution and solid matrix primed with Micro Cel E (seed, Micro Cel E, and water mixed with the ratio of 4.0:2.0:8.0 by weight for carrot, and the ratio of 4.0:1.2:5.0 for onion at $20^{\circ}C$ for 4 days). Seeds were rapidly absorbed water after osmotic priming and solid matrix priming treatment. The moisture content of solid matrix primed seeds was lower than that of osmotic primed seeds in carrot and onion. Osmotic priming and solid matrix priming did not influenced on increasing of percent germination, but $T_{50}$ of treated seeds was shorter than those of untreated seeds at four temperature regimes. The effects of osmotic or solid matrix priming in reducing the $T_{50}$ was greater when the seeds were germinated at $15^{\circ}C$ than at $25^{\circ}C$ and $30^{\circ}C$ temperature. Solid matrix primed seeds germinated faster than osmotic primed seeds at all temperature in both carrot and onion. After priming, surface-dried seeds germinated faster than dried-back seeds in carrot and onion. Emergence speed of carrot and onion seeds was faster than untreated seeds by osmotic priming or SMP treatment although the final emergence percentage was not significantly influenced. On the other hand, early growth was not significantly influenced by osmotic priming or SMP treatment of carrot and onion seeds.

Preparation and Characterization of Antimicrobial Films Using Water Soluble Polymer (수용성 고분자를 이용한 항균 필름의 제조 및 특성 연구)

  • Choi, Jun Ho;Choi, Yoo Sung;Oh, Il Hong;Kim, Maeng Su;Lee, In Hwa
    • Korean Chemical Engineering Research
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    • v.49 no.4
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    • pp.449-455
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    • 2011
  • This study was performed to develop antimicrobial films using polyvinyl alcohol and methyl cellulose. Methyl cellulose and polyvinyl alcohol films plasticized with PEG(polyethylene glycol) were prepared by solvent casting process under addition of 0.025~1.0 wt% ampicillin and 0.1~1.0 wt% streptomycin as an antimicrobial agent. The mechanical properties of prepared films were examined by universal testing machine(UTM). Tensile strength of methyl cellulose films was 15.44~21.70 $N/mm^2$. Tensile strength of PVA(15 wt%) film was 20.2~51.5 $N/mm^2$, and the tensile strength of the antimicrobial films were decreased linearly with increasing the antibiotic loading amount up to 1 wt%. Antimicrobial activities of PVA and methyl cellulose films containing ampicillin and streptomycin through the disc diffusion test for the Staphylococcus aureus and Escherichia coli. The antimicrobial activity of methyl cellulose films and PVA containing ampicillin were higher than that of containing streptomycin methyl cellulose films. The results indicate the films may be a proper materials for antimicrobial packing applications.

Dehydration and Drying Characteristics of Gingers Using Dehydrating Agent by Dextrose Equivalent and Molecular Weight Condition (포도당 당량과 분자량 조건별 탈수제를 적용한 생강의 탈수와 건조 특성)

  • Lee, Hyun-Seok;Kwon, Ki-Hyun;Kim, Byeong-Sam;Jeong, Jin-Woong;Kim, Jong-Hoon;Sung, Jung-Min
    • Food Science and Preservation
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    • v.17 no.6
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    • pp.763-769
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    • 2010
  • We examined variations in ginger dehydration and recovery rate upon use of dextrose of different equivalence values. The dehydration rate varied with dextrose equivalent and the dehydration rate increased as the equivalence value increased. Both dehydration and recovery rates varied with dextrose molecular weight. Moisture content was lowest in samples prepared by freeze-drying, and neither dextrose equivalent nor molecular weight affected moisture level. Upon color analysis, ginger dried using dextrose varying in equivalence and molecular weight was similar in color to the original material, unlike ginger dried by other methods. Hot-air-dried ginger scored lowest in all sensory tests, compared with ginger prepared by molecular press dehydration using dextrose varying in equivalence and molecular weight. With respect to the appearance of ginger, freeze-dried samples were optimal, but molecular press dehydration yielded samples that scored best upon overall evaluation. When all quality evaluation items were taken together, molecular press dehydration resulted in a better quality product than the older hot-air or freeze-drying methods.

Studies on Protoplast Formation and Regeneration of Coriolus versicolor (구름버섯의 원형질체(原形質體) 형성(形成)과 재생(再生)에 관한 연구(硏究))

  • Bok, Jin-Woo;Park, Seol-Hee;Choi, Eung-Chil;Kim, Byong-Kak;Yoo, Young-Bok
    • The Korean Journal of Mycology
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    • v.18 no.3
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    • pp.115-126
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    • 1990
  • To establish basic techniques for protoplast fusion of Coriolus versicolor several factors affecting protoplast formation and regeneration were investigated. Protoplast isolation was at maximum with 2.5-day cultured mycelia of C. versicolor treated with the combination of two enzymes, Novozym 234 (10 mg/ml) and cellulase Onozuka R-10 (15 mg/ml), for 3-4.5 hours at $30^{\circ}C.$ As an osmotic stabilizer for stabilizing the protoplast, 0.6 M sucrose was the best for formation and regeneration of the protoplast from the mycelia of the fungus and the regeneration frequency was 3.48%. Protoplast fusion was made by a modified method of Peberdy using PEG (M.W. 4,000). The fusion frequency between two mutants of C. versicolor was 1.86% and the fusion products showed differences in growth rate and colony morphology.

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