• Title/Summary/Keyword: Mono Q

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Production and Purification of Soluble Recombinant Human Lymphotoxin in Escherichia coli

  • Choi, Yoon-Ho;Na, Doe-Sun;Pan, Jae-Gu;Park, Seung-Kiel;Yoo, Hyang-Sook;Kang, Kook-Hee;Sung, Moon-Hee
    • Journal of Microbiology and Biotechnology
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    • v.2 no.1
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    • pp.21-25
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    • 1992
  • Human lymphotoxin (LT) was produced in E. coli as a soluble protein. The level of recombinant human LT production was about 4% of the total souble proteins of E. coli extracts. Recominant human LT was purified to apparant homogeneity by a simple procedure utilizing FPLC on Mono Q and Mono S columns. The specific activity of the purified LT was $10\times10^7\;units/mg$.

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Purification and Characterization of Polyphenol Oxidase from Lotus Root (Nelumbo nucifera G.) (연근의 polyphenol oxidase 정제 및 특성조사)

  • Moon, Sang-Mi;Kim, Hyun-Jin;Ham, Kyung-Sik
    • Korean Journal of Food Science and Technology
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    • v.35 no.5
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    • pp.791-796
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    • 2003
  • Polyphenol oxidase isoforms were purified from the lotus roots using 50% acetone precipitation, conventional chromatographies of Q-Sepharose and hydrophobic interaction, and high performance liquid chromatographies of Mono-Q and Superdex 75 gel-filtration. Molecular mass of a purified PPO isoform (LPIII-2) was determined to be 56 kDa using gel-filtration chromatography. The active form of LPIII-2 appeared to bea heterodimer, as purified LPIII-2 on SDS-PAGE gel showed two bands that were determined to be 28 kDa and 26 kDa. To further characterize PPO, partially purified PPO isoforms (LP-II, LP-III) were obtained from Q-Sepharose anion-exchange chromatography. In substrate specificity, the partially purified PPO isoform LP-II showed a high affinity to catechol, while LP-III showed a high affinity to pyrogallol. The optimum pH of LP-II and LP-III was pH 7.0. Interestingly, the partially purified PPO isoforms showed high activities at low temperatures $(0{\sim}5^{\circ}C)$, and as temperatures rose, the activities decreased. Both PPO isoforms were stable at $40^{\circ}C$ and were inactivated by incubation at $60^{\circ}C$ for 40 min.

Purification and Assay of Extracellular Autolysin from Moraxella sp. CK-l (Moraxella sp. CK-1의 세포외 Autolysin의 분리 정제 및 활성도 측정)

  • 오영상;이장현;한명수;윤문영
    • Korean Journal of Microbiology
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    • v.39 no.3
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    • pp.148-154
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    • 2003
  • Moraxella sp. CK-l is known to inhibits the growth of Anabaena cylindrica, a cyanobacterium. It has been documented that the ability of this growth inhibition of Anabaena cylindrica was attributed to extracellular autolysin from Moraxella sp. CK-l. However, it remains to be elucidated identification and characterization of autolysin have yet been elucidated. In this study, we tried to purify and identify autolysin secreted from Moraxella sp. CK-l. Cells were grown in a complex liquid medium (BGC-11) and culture supernatants were collected, followed by ammonium sulfate fractionation. Fractions were further separated with anion exchange column, Mono-Q, in FPLC system and analyzed by SDS/PAGE. The fraction containing high autolysin activity showed a single distinct protein peak in anion column and molecular mass of about 17 kDa in SDS/PAGE. Nterminal amino acid sequencing of the protein was analyzed, of which result showed the homology with some proteases, including extracellular serine protease, Dichelobacter nodosus.

Trends of Post-fire Forest Recovery in the South Sikhote-Alin Mountains, Russian Far East

  • Komarova, Tatiana A.;Sibirina, L.A.;Papaik, M.J.;Park, J.H.;Kang, HoSang
    • Journal of the Korean Society of Environmental Restoration Technology
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    • v.16 no.3
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    • pp.83-95
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    • 2013
  • To understand natural regeneration and stand development after fire in mixed broadleaved-coniferous forests of Sikhote-Alin Mountains, ten sample plots of $50m{\times}50m$ size were established in 1975 and 1983 at the stands burned by wildfires in 1973 and 1982, respectively. And, the number of naturally regenerated seedlings were monitored in two $50m{\times}4m$ subplots in each plot. The most fire-sensitive conifer species is Abies nephrolepis, while Betula costata is the most fire-sensitive broadleaved tree species. The most fire-resistant species were Q. mongolica, T. taquetii and A. mono. The results of 20 and 30 years after the fire showed that pioneer tree species, e.g. Populus, Salix, and Betula, were regenerated immediately at the early stage of stand development and grew where there is a mono canopy layer with high density. On the other hand, the densities of successors, e.g. Pinus koraiensis, Picea jezoensis, Abies nephrolepis, Acer mono and Tilia taquetii, which were present in the study plots before the fire, increased gradually. Naturally regenerated tree species after forest fire by the growth rate were divided into three groups according to their annual height growth. The seral tree species (Betula costata, Betula platyphylla, Padus maackii, Populus tremula and Sarix caprea) belong to the first group and have the highest growth rate (from 40 to 96 cm per year). The late successional broad-leaved trees (Tilia taquetii, Acer mono and Quercus mongolica) belong to the second group and have intermediate annual height growth (from 3.7 to 13.5 cm per year). The late successional coniferous species (Picea jezoensis, Pinus koraiensis and Abies nephrolepis) form the third group and have the least annual height growth (from 1.4 to 3.5 cm per year).

Purification and Characterization of the Recombinant Arabidopsis thaliana Acetolactate Synthase

  • 조규봉;홍성택;최명운;장수익;최정도;고은희
    • Bulletin of the Korean Chemical Society
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    • v.18 no.6
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    • pp.648-653
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    • 1997
  • Acetolactate synthase was purified from Escherichia coli MF2000/pTATX containing Arabidopsis thaliana acetolactate synthase gene. Purification steps included DEAE cellulose ion exchange column chromatography, phenyl sepharose hydrophobic column chromatography, hydroxylapatite affinity column chromatography, and Mono-Q HPLC. Molecular weight was estimated to be ∼65 KDa and purification fold was 109 times. The enzyme showed a pH optimum of 7 and the $K_M$ value was 5.9 mM. The purified enzyme was not inhibited by any of the end products, valine, leucine, and isoleucine.

A Study on the Menopause Experience : A Q Methodological Approach (폐경기 경험에 관한 연구 : Q 방법론적 접근)

  • 신혜숙
    • Journal of Korean Academy of Nursing
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    • v.25 no.4
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    • pp.807-824
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    • 1995
  • This study was intended to provide women who are experiencing menopause with effective nursing care by exploring the menopause experience. The purpose of the research was to understand the subjective feelings of women about the question of what the menopause experience is. Q-Methodological method was used for that purpose. As the research method, Q-statements were collected preliminary to the study of through in -depth interviews and a literature review. For the study 34 Q-statements were selected. There were 21 women as subjects for the research. The 21 women sorted the 34 statements using the principle of Forced Normal Distribution. The principle of Forced Normal Distribution, which has nine scales to measure the individual opinions, was called Q-Factor Analysis by using PC Quanl Program to supply the material. As a result, there were four categories(self-com-passion type, self-regulation type, self-perception type, self-abandonment type) of special opinion about the menopause experience in these women. The first type was called Self-compassion This type' was associated with varying degrees of emotonal instability(psychological withdrawal). And type in menopause signifies loss of a socially valued status and may result in depressive symptoms. This type expresses the menopause experience as associated with negative reaction. The second type is called Self-regulation. This type overcomes the menopause experience more actively than the other types and do not express the menopause experience as one of suffering. This type make efforts by themselves to regulate the mono-pause experience. The third type is called Self-perception. This type perceives the experience of menopause which as typical menopausal syndrome (hot flashes and decreased vaginal lubrication, decreased estrogen producing atrophic changes of the labia and vaginal mucosa, making intercourse uncomfortable). The fourth type is called Self- abandonment. This type denies the experience of menopause. The meaning of menopausal experience is significantly related with a life accident or life load. Also, This type experiences pain in the musculoskeletal system. As a result, The meaning of the menopausal experience is affected by perceived subjective experience of the nurses, the need to understand each persons meaning of the menopause experience and to develop appropriate nursing interventions based on the typology of menopause experience. Finally. The result of the study will provide basic data for nursing intervention the menopausal women.

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Characteristics of a New Obligate Methanol-Oxidizing Bacterium (새로운 절대 메탄올 산화세균의 분리 및 특성)

  • Kim, Si-Wook;Park, Yong-Ha
    • Korean Journal of Microbiology
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    • v.31 no.4
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    • pp.261-266
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    • 1993
  • A new methyltrophic bacterium which utilizes methanol as a sole source of carbon and energy was isolated from soil. It was Gram-negative, nonmotile, nonspore-forming rod, and strictly aerobic bacterium. Catalase and oxidase activities were present. Nitrate was reduced to nitrite. Vitamins and other growth factors were not required. Generation time was 1.6 hr under the optimal condition. The isolate assimilated methanol via the ribulose mono-phosphate pathway (Enter-Doudoroff varient) and did not have .alpha.-ketoglutarate dehydrogenase. It assimilated ammonia through glutamate dehydrogenase. The guanine plus cytosine content of the DNA was 61.0 mol%. The celular fatty acid composition was primarily straight-chain saturated $C^{16 : 0}$ acids (palmitic acids) and unsaturated $C_{16 :1}$ acid (palmitoleic acids), and the isolate also contained two unidentified $C_{17}$ branched fatty acids. The major ubiquinone was Q-8, and Q-6 and Q-7 were present as minor components. Phosphatidylethanolamine and phosphatidylglycerol were predominantly present, and diphosphatidyglycerol was also detected. Based on the physiological and biochemical properties, the isolate was assigned to a novel species of the genus Methylobacillus, Methylobacillus methanolovorus sp. nov.

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Purification and Characterization of Mitogen -Activated Protein (MAP) Kinase from Mammalian Tissue Cells (동물 조직세포로부터 Mitogen-activated Protein (MAP) Kinase의 분리 및 성격규명)

  • 김태우;정동주;김윤석
    • Biomedical Science Letters
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    • v.2 no.1
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    • pp.21-30
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    • 1996
  • MAP kinases are a family of serine/threonine specific protein kinases becoming activated in response to different proliferative stimuli by phosphorylation at both threonine and tyrosine residue. Present study shows that MAP kinase was purified from P388 murine leukema cells by SP sephadex C-50, phenyl superose and Mono Q column chromatography and identified with anti-ERKl antibody by western blotting. Immnublotting analysis to the crude extract of P388 cell lysate shows 44 kD and other minor bands but partial purified fraction eluted from phenyl supherose column have 44kD and 66 kD isoform. Subcloned GST-fusion protein from N-terminal of $p56^{kk}$ was tested as a substrate for MAP kinase phosphorylation. It was showed that the wild type and mutant forms(S42A) were fully phosporylated by purified MAP kinase fraction as com-pare with the other mutant form(S59A). This finding suggest that those GST-fusion proteins may be used as substrate for the in vitro test of MAP kinase.

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N-terminal amino acid sequence analysis of major manganese peroxidase (MnP3) produced by static culture of Pleurotus ostreatus (느타리 버섯균의 정치배양으로부터 생산되는 중요한 망간퍼옥시데이즈(MnP3)의 N-말단 아미노산 배열 분석)

  • Ha, Hyo-Cheol
    • Journal of Mushroom
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    • v.17 no.4
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    • pp.185-190
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    • 2019
  • Pleurotus ostreatus No.42, known as the ligninolytic basidiomycetes, showed production of MnP and Lac, but did not show any LiP acitivity in static culture, grown in GPYW liquid medium. Maximum production of MnP (80U/flask) was observed on day 11 of culturing in this medium. Chromatographic purification of MnP included the use of Sepharose CL-6B and Mono-Q. The major MnP isozyme purified by column chromatography was observed to be a 36.4 KDa (single band on SDS PAGE). The 19-amino acid sequence from the N-terminal was determined by protein sequencing to be ATCADGRTTANAACCVLFP. The N-terminal sequence of the major MnP isozyme of P. ostreatus No.42 was found to be the same as a previously reported sequence of an MnP3 isozyme from this fungus.

Effect of Number of Measurement Points on Accuracy of Muscle T2 Calculations

  • Tawara, Noriyuki;Nishiyama, Atsushi
    • Investigative Magnetic Resonance Imaging
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    • v.20 no.4
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    • pp.207-214
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    • 2016
  • Purpose: The purpose of this study was to investigate the effect of the number of measurement points on the calculation of transverse relaxation time (T2) with a focus on muscle T2. Materials and Methods: This study assumed that muscle T2 was comprised of a single component. Two phantom types were measured, 1 each for long ("phantom") and short T2 ("polyvinyl alcohol gel"). Right calf muscle T2 measurements were conducted in 9 healthy male volunteers using multiple-spin-echo magnetic resonance imaging. For phantoms and muscle (medial gastrocnemius), 5 regions of interests were selected. All region of interest values were expressed as the mean ${\pm}$ standard deviation. The T2 effective signal-ratio characteristics were used as an index to evaluate the magnetic resonance image quality for the calculation of T2 from T2-weighted images. The T2 accuracy was evaluated to determine the T2 reproducibility and the goodness-of-fit from the probability Q. Results: For the phantom and polyvinyl alcohol gel, the standard deviation of the magnetic resonance image signal at each echo time was narrow and mono-exponential, which caused large variations in the muscle T2 decay curves. The T2 effective signal-ratio change varied with T2, with the greatest decreases apparent for a short T2. There were no significant differences in T2 reproducibility when > 3 measurement points were used. There were no significant differences in goodness-of-fit when > 6 measurement points were used. Although the measurement point evaluations were stable when > 3 measurement points were used, calculation of T2 using 4 measurement points had the highest accuracy according to the goodness-of-fit. Even if the number of measurement points was increased, there was little improvement in the probability Q. Conclusion: Four measurement points gave excellent reproducibility and goodness-of-fit when muscle T2 was considered mono-exponential.