• Title/Summary/Keyword: polyacrylamide,

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Studies on the Intraspecific Variations on Geological Distributions of Acusta despecta sieboldiana in Korea (한국산 달팽이 ( Acusta despecta sieboldiana) 의 지리적 분포에 따른 종내 변이에 관한 연구)

  • 이준상;권오길
    • The Korean Journal of Malacology
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    • v.12 no.1
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    • pp.19-31
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    • 1996
  • 국내 11개 지역(경기도 옹진군 덕적도, 경기도 의정부시 소요산, 강원도 춘천시, 충청남도 태안군 안홍, 경상북도 울릉군 울릉도, 전라북도 부안군 변산, 전라남도 신안군 홍도, 흑산도 비금도, 진도, 제주도)에서 채집된 달팽이 (A. despecta sieboleiana)를 대상으로 외부형태 분석과 동위효소를 검출하여 각 지역 집단간의 형태적, 유전적 유연관계와 변이 정도를 연구한 결과는 다음과 같다. 외부형태에 의한 집단간 유연관계는 안홍과 덕적도집단(average taxonomic distance, D=0.358)이 형태적으로 가장 유사한 집단으로 나타났다. Polyacrylamide gel을 사용하여 11가지의 동위효소를 검출한 결과 AKP, ACP, AO, EST, GPD, HBDH, LDH, SDH, XDH의 동위효소에서 유전적 다형현상이 나타났다. 전 집단의 평균 다형형의 빈도는 59.19%이며 이형접합자 빈도는 0.263이고 각 집단의 유전적 차이치는 춘천과 울릉도 집단이 0.066(genetic distance)으로 가장 높은 유사성을 보였다. 지리적 분포에 따른 외부형태 변이와 유전적 변이와의 관계는 유의성이 없었다.

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Possible Roles of LAMMER Kinase Lkh1 in Fission Yeast by Comparative Proteome Analysis

  • Cho, Soo-Jin;Kim, Young-Hwan;Park, Hee-Moon;Shin, Kwang-Soo
    • Mycobiology
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    • v.38 no.2
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    • pp.108-112
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    • 2010
  • To investigate the possible roles of LAMMER kinase homologue, Lkh1, in Schizosaccharomyces pombe, whole proteins were extracted from wild type and lkh1-deletion mutant cells and subjected to polyacrylamide gel electrophoresis. Differentially expressed proteins were identified by tandem mass spectrometry (MS/MS) and were compared with a protein database. In whole-cell extracts, 10 proteins were up-regulated and 9 proteins were down-regulated in the mutant. In extracellular preparations, 6 proteins were up-regulated in the lkh1+ null mutant and 4 proteins successfully identified: glycolipid anchored surface precursor, $\beta$-glucosidase (Psu1), cell surface protein, glucan 1,3-$\beta$-glucosidase (Bgl2), and exo-1,3 $\beta$-glucanase (Exg1). These results suggest that Lkh1 is involved in regulating cell wall assembly.

Simulation of Alpine Field Soil Loss by Outdoor Rainfall Simulator (실외인공강우에 의한 경사밭의 토양 유실량 모의)

  • Shin, Hyun-Jun;Won, Chul-Hee;Kim, Tae-Yoo;Choi, Eu-Tteum;Choi, Joong-Dae
    • Proceedings of the Korea Water Resources Association Conference
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    • 2010.05a
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    • pp.1242-1246
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    • 2010
  • 호우기간 동안 내리는 빗방울의 타격에 의해 흙 입자는 침출수와 함께 이동하여 지표 아래 공극을 막는다. 다져진 지표면은 유출과 토양 유실의 원인이 된다. 발생원으로부터 유실되는 토양을 Polyacrylamide(PAM)과 지표피복재를 이용하여 저감하는 연구를 하였다. PAM은 토양 입자의 결합력을 강화시키고 이탈을 방지하여 토양 유실을 감소시키는데 효과적이다. 이 연구의 목적은 PAM을 덧붙인 볏짚거적, 볏짚거적+톱밥, 볏짚거적+왕겨 등을 이용하여 인공강우 동안 토양 유실을 저감하는 효과를 조사하는 것이다. 실험은 1시간 동안 강우를 모의했으며, 실험 도중에 6~7회 수질 샘플을 채취하였다. 초기유출시간은 총 4차 실험중에 2차를 제외한 나머지에서 볏짚거적+왕겨+PAM이 가장 느리게 유출되었다. SS와 탁도 항목에서 1차 실험을 제외한 나머지 실험에서 볏짚거적+톱밥+PAM이 효과적이었다. 단순히 PAM을 사용하는 것보다 잔여물(residue)을 같이 혼합한 피복재료가 토양 유실과 유출을 저감하는데 더욱 효과적인 것으로 나타났다. 본 연구 결과는 향후 농촌지역에 토양 유실저감에 필요한 자료로 활용할 수 있을 것이라 판단된다.

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Purification and Acetylation of Protein X Subunit of Pyruvate Dehydrogenase Complex (PDC) from Bovine Kidney

  • Ryu, Ryu;Song, Byoung-J.;Hong, Sung-Youl;Huh, Jae-Wook
    • Archives of Pharmacal Research
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    • v.19 no.6
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    • pp.502-506
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    • 1996
  • Protein X is one of the subunits of pyruvate dehydrogenase complex. The biological role of this protein has not been fully elucidated, mainly because of the difficulty in its dissociation from the tightly bound dihydrolipoamide acetyltransferase-protein X subcomplex. We have found that the detachment of protein X from acetyltransferase subunit can be easily accomplished by the cycles of freezing and thawing proces. Several lines of evidence including sodium dodecyl sulfate-polyacrylamide gel electrophoresis, N-terminal amino acid sequence analysis and acetylation with $[2^{14}C]$ pyruvate confirmed that the purified protein is protein X. The purified intact form of protein X was acetylated by $[2^{14}C]$ pyruvate in the presence of py-ruvate dehydrogenase subunit.The acetylation efficiency of this protein was lower than that of acetyltransferase and was not affected by the presence of acetyltransferase.

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Nucleotide Sequence and Homology Analysis of phnC Gene Encoding Glutathione S-transferase from Pseudomonas sp.DJ77 (Pseudomonas sp. DJ77에서 Glutathione S-transferase를 암호하는 phnC 유전자의 염기서열과 상동성 분석)

  • 우희종;신명수;김성재;정용제;정안식;박광균;김영창
    • Korean Journal of Microbiology
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    • v.33 no.2
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    • pp.86-91
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    • 1997
  • Pseudomonas sp. DJ77로부터 클로닝된 glutathione S-transferase 유전자(phnC)의 염기서열을 결정하였다. 603bp의 open reading frame(ORF)이 존재하였고 개시코돈 앞에서 Shine-Dalgarno sequence를, 종결코돈 뒤에서는 terminator sequence를 발견하였다. phnC 유전자에서 만들어지는 phnC 단백질은 21,416 Da으로 SDS-polyacrylamide gel 전기영동 결과와 일치하였다. PhnC는 Bulkholderia cepacia LB400, Cycloclasticus oligotrophus RB1의 GST와 각각 53.7%, 49%의 높은 상동성을 나타냈다. 아미노산 서열의 상동성과 필수잔기들의 존재유무로 판단할 때 PhnC GST는 theta class GSTs와 진화적으로 유연관계가 높았지만 alpha, mu, pi, sigma class GSTs에서 구조적, 기능적으로 중요하다고 알려진 아미노산 잔기들이 PhnC GST에도 보존되어 있었다. 또한, phnC 유전자의 위치가 C. oligotrophus RB1, B. cepacia LB400 등의 GST 유전자 위치와 유사하다는 점에서 PhnC 효소는 난분해성 방향족 탄화수소의 분해에 관여하는 것으로 생각된다.

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Proteome Analysis of Amniotic Fluid by gradient 2-D PAGI (Gradient 2-D PAGE를 이용한 양수 프로테옴 분석)

  • 이은희;김재찬;변상요
    • KSBB Journal
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    • v.18 no.1
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    • pp.35-38
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    • 2003
  • Analysis of proteome in amniotic fluid was performed by 2-D PAGE (polyacrylamide gel electrophoresis). Proteins in amniotic fluid were separated by centrifugation and solubilized in buffer solution for IEF, using an IPG strip of pH 4-7L. Both a homogeneous slab gel of 12.5% and a gradient gel of 8-18%, were used. After 2-D PAGE, spots were stained with silver nitrate and picked up for in-gel digestion. Digested peptides were analyzed by MALDI-TOF and proteins were further identifical. More protein spots were detected in the gradient gels and a protein not previously reported was identified.

Purification and Characterization of Xylanase from Bacillus sp. Strain DSNC 101

  • Cho, Nam-Chul;Bai, Suk
    • Journal of Microbiology and Biotechnology
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    • v.7 no.6
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    • pp.386-390
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    • 1997
  • A xylanase from the Bacillus sp. strain DSNC 101, isolated from soil, was purified to homogeneity by anion-exchange and hydrophobic interaction chromatography followed by gel filtration chromatography. The enzyme cleaved xylan, but not carboxymethyl cellulose, Avicel, soluble starch, and pNPX. The main product of oat spelts xylan hydrolysates was xylobiose. The xylanase had a molecular weight of 25 kDa determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Optimum temperature and pH for the xylanase activity were $50^{\circ}C$ and 6.0, respectively. $K_{m}\;and\;V_{max}$ of the enzyme for oat spelts xylan were 12.5 mg of xylan/ml and 869.5 unit/mg of protein, respectively. Xylanase was completely inhibited by Hg, Cu, and N-bromosuccinimide, but was stimulated by Ca, Co, and Mg.

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Purification and Characterization of Cycloinulooligosaccharide Fructanotransferase from Bacillus macerans CFC1

  • Kim, Hwa-Young;Choi, Yong-Jin
    • Journal of Microbiology and Biotechnology
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    • v.8 no.3
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    • pp.251-257
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    • 1998
  • Cycloinulooligosaccharide fructanotransferase (CFTase) which produces cyclofructan from inulin was purified 332-fold from a culture broth of Bacillus macerans CFCl. The molecular mass of the CFTase was estimated to be 110 kDa by SDS-polyacrylamide gel electrophoresis and gel filtration, indicating that the enzyme has a monomer structure. The maximal level of enzyme activity was observed at pH 7.5 and $45^{\circ}C$. The enzyme was stable in the pH range 6.0 to 9.5, and at temperatures up to $45^{\circ}C$ for 1 h. The enzyme activity was completely inhibited in the presence of 0.5 mM $Ag^+\;or\;Cu^2+$ ion. None of sucrose (GF), l-kestose (GF2), or nystose (GF3) were found to be substrates for the CFTase, but inulooligosaccharides larger than nystose were attacked by the enzyme. The CFTase catalyzes not only the cyclization as the major reaction, but also disproportionation and coupling reactions involving intermolecular transfructosylation in the same manner as cyclodextrin glucanotransferase (CGTase) (EC 2.4.1.19).

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A study on the optimal HPLC condition for peptides complex analysis using mass spectrometry (질량분석기를 단백질 분석에 적용하기 위한 고성능액체크로마토그래피 최적조건 연구)

  • Kwon, Sung Won;Park, Chul Hong
    • Analytical Science and Technology
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    • v.16 no.1
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    • pp.78-81
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    • 2003
  • Peptides separation in high performance liquid chromatography (HPLC) is very important for the analysis of total proteins using mass spectrometry rather than two-dimensional polyacrylamide gel electrophoresis (2D-PAGE). In this study, we investigated the optimal HPLC condition of peptides for the use of mass fingerprinting. As a result of pursuing a combination of solvent additives for HPLC, water and acetonitile containing both 0.1% trifluoroacetic acid and 0.1% acetic acid respectively showed the most efficient resolution and sensitivity.

An Integrated Process for the Separation and Purification of Biologically Active Proteins from Human Urine (인뇨로부터 유용단백질의 통합 분리정제 공정)

  • 김기용;정광회문흥모
    • KSBB Journal
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    • v.8 no.3
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    • pp.287-294
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    • 1993
  • For the purpose of combining the purification processes for several biologically active proteins form human urine, an efficient integrated fractionation procedure has been investigated. The procedure was started by concentration with ultrafiltration and pH precipitation followed by a selectable combination of chromatography on gel filtration, adsorption, ion exchanger, affinity, and reverse phase column. By this process, the purified urokinase, epidermal growth factor and albumin migrated as a single band on SDS-polyacrylamide gel electrophoresis and were fully active. The recoveries of these purified proteins were 48%, 17%, and 46%, respectively.

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