• 제목/요약/키워드: 2D gel electrophoresis

검색결과 335건 처리시간 0.023초

Purification and Characterization of Inulin Fructotransferase (Depolymerizing) from Arthrobacter sp. A-6

  • PARK, JEONG-BOK;YONG-JIN CHOI
    • Journal of Microbiology and Biotechnology
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    • 제6권6호
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    • pp.402-406
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    • 1996
  • Inulin fructotransferase (depolymerizing) (EC 2.4.1.93) was purified 34-fold from the culture broth of Arthrobacter sp. A-6 by using a combination of ammonium sulfate fractionation, DEAE-Sepharose CL-6B chromatography and Sephacryl S-200 gel filtration. The purified enzyme converts inulin into di-D-fructofuranose dianhydride III(DFA III) and small quantities of fructo-oligosaccharides. The temperature and pH optima of the enzyme were $70^{\circ}C$ and 6.0, respectively. Molecular weight of the enzyme was determined to be 49 kDa by 12$%$ SDS-polyacrylamide gel electrophoresis, and 145 kDa by Sephacryl S-200gel filtration. This indicates that the functional inulin fructotransferase of Arthrobacter sp. A-6 has a homomeric trimer structure. The enzyme had an isoelectric point of pH 4.6. The N-terminal amino acid sequence of the purified enzyme subunit was Ala-Asp-Asn-Pro-Asp-Gly(\ulcorner)-Ser-Asn-Met(or Glu)-Tyr-Asp-Val.

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하천에서 분리한 비소 내성세균의 유전적 특성 (Genetic Characteristics of Arsenic Compounds-Resistant Bacteria Isolated from Stream Water)

  • 정미경;이호자
    • 미생물학회지
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    • 제29권1호
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    • pp.63-68
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    • 1991
  • Several arsenic compound-resistant bacteria were isolated from Jung-Rang stream. The isolates, D-3, D-12, and D-14 were characterized phenotypically and genetically, and identified as Serratia liquefaciens, Klebsiella oxytoca, and Klebsiella pneumoniae, respectively. A plasmid of 67kb was found in Klebsiella oxytoca D-12 and designated as pMH12. Transfer of this plasmid from D-12 to E. coli HB101 was occurred, and the resulting transconjugant strains expressed the same level of heavy metal resistance as the donor strain. The physical presence of this plasmid in transconjugant was detected with agarose gel electrophoresis. Arsenite-sensitive derivatives of isolate D-12 were obtained with Mitomycin C treatment which cured pMH12. Antibiotics and heavy metal resistances were also examined to be used as a proper marker for the isolates in gene cloning. Isolate D-12 has resistance to several heavy metal ions such as $Cd^{2+}$ , $Zn^{2+}$ and $Hg^{ 2+}$ Also, all the other arsenite resistant isolates showed resistance to several heavy metal ions and antibiotics.

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재래한우의 혈액단백질 및 효소의 유전적 다형현상 (Genetic Polymorphisms of Blood Proteins and Enzymes in Korean Native Cattle)

  • 상병찬;류승희;이상훈;한성욱;이창수
    • 농업과학연구
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    • 제22권2호
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    • pp.170-179
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    • 1995
  • 본 연구는 축협중앙회 한우개량사업소에서 사육하고 있는 한우 238두의 혈청내에 존재하는 혈액 단백질 및 효소의 유전적 다형을 조사하기 위하여, 혈청단백질인 post-transferrin-2(pTf-2), transferrin(Tf), post-albumin(pAlb) 및 albumin(Alb)과 혈청 효소인 ceruloplasmin(Cp) 및 amylase-I(Am-I), 그리고 혈구단백질인 hemoglobin(Hb)을 PAGE(polyacrylamide gel electrophoresis)와 STAGE(starch gel electrophoresis) 방법으로 유전적 다형을 분석하여 이들의 유전자형 및 유전자빈도를 추정하였던바 그 결과는 다음과 같다. 1. 혈청단백질인 pTf-2 좌위는 pTf-2 F와 S의 공우성 대립유전자가 검출되었고, 유전자형은 pTf-2 FF, FS 및 SS 형이 확인되었으며, 이들의 분포는 각각 46.22, 46.64 및 7.14%이었고, 유전자 빈도는 pTf-2 F와 S에서 각각 0.695 및 0.305이었다. 2. Tf 좌위는 Tf A, D1, D2 및 E의 대립유전자가 검출되었으며, 유전자형에 있어서는 Tf AA, ADl, AD2, AE, D1D1, D1D2, D2D2 DIE D2E 및 EE형이 확인되었고, 이들의 분포는 각각 0.84, 13.87, 13.03, 10.92, 22.27, 12.61, 2.94, 15.l3, 6.72 및 1.68%이었으며, 유전자빈도에 있어서는 Tf A, D1, D2 및 E의 빈도가 각각 0.197, 0.430, 0.191 및 0.181이었다. 3. pAlb 좌위는 pAlb F와 S 유전자가 검출되었으며, 유전자형의 분포에 있어서는 pAlb FFFS 및 SS 형이 각각 42.86, 33.19 및 23.95%이었고, 유전자빈도에 있어서는 pAlb F와 S에서 각각 0.595 및 0.405이었으며, Alb에서는 Alb A 유전자 빈도가 1.000이었다. 4. Cp 좌위는 Cp F와 S 유전자가 확인되었으며, 유전자형의 분포는 Cp FF, FS 및 SS 형에서 각각 23.11, 34.87 및 42.02% 이었고, Cp F와 S 유전자빈도는 각각 0.405 및 0.595이었다. 5. Am-I 좌위는 Am-I B와 C 대립유전자가 검출되었으며, 유전자형의 분포는 Am-I BB, BC 및 CC형에서 각각 51.26, 16.81 및 31.93%이었고, 유전자빈도는 Am-I B와 C가 각각 0.597 및 0.403이었다. 6. Hb 좌위는 Hb A와 B 대립유전자가 검출되었으며, 유전자형의 분포는 Hb AA, AB 및 BB형에서 각각 83.19, 16.39 및 0.42%이었고, Hb A와 B의 유전자빈도는 각각 0.914 및 0.086%이었다.

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Disassembly of Chlorophyll-Protein Complexes in Arabidopsis thaliana during Dark-Induced Foliar Senescence

  • Lee, Choon-Hwan
    • Journal of Plant Biology
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    • 제39권4호
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    • pp.301-307
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    • 1996
  • The disassembly of Chl-protein complexes during dark-induced senescence (DIS) was investigated using detached third and fourthleaves of 21$\pm$1 day-old Arabidopsis thaliana. Although Chl content decreased linearly after 1 d, a significant decrease of photochemical effeciency (Fv/Fm) was observed after 2 d. In experiments using native green gel electrophoresis of Chl-protein complexes combined with additional two-dimensional SDS-PAGE analysis, we could observe the degradation of both photosystems after 2 d. Although light-harvesting complex(LHC) for PSI (LHCI) was degraded first in PSI complex, small PSII apoproteins including CP47/CP43 and D1/D2 apoproteins were degraded first in PSII complexes. LHC for PSII (LHCII) trimers were stable until 4 d. The level of LHCII monomers was increased until 3 and decreased thereafter, resulting in the increase of free pigments. These results suggest that the disassembly process of PSI is different from that of PSII.

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Proteomic Analysis기법을 이용한 홍화자약침액(紅花子藥鍼液)이 간암세포주(肝巖細胞柱)의 유전자(遺傳子) 발현(發顯)에 미치는 영향(影響) (Effect of Carthami Tinctorii Fructus Herbal-acupuncture Solution(CTF-HAS) on Gene Expression in HepG2 carcinomar cells by Proteomic Analysis)

  • 이경민;임성철;정태영;서정철;한상원
    • 대한약침학회지
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    • 제8권2호
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    • pp.23-28
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    • 2005
  • Objective : It has long been known about the osteogenic effect of CTF-HAS on bone tissues. However, it has not been determined the effect of CTF-HAS on cancer cells. The purpose of this study is to screen the CTF-HAS mediated differentially expressed genes in cancer cells such as HepG2 hepatoma cells lines. Methods : CTF-HAS was prepared by boiling and stored at $-70^{\circ}C$ until use. For proteomic analysis, total protein was analyzed by 2D gel electrophoresis and Q-TOF mass spectrometer. Results : In proteomic analysis, three spots were identified by 2D-gel electrophoresis and Q-TOF analysis. One down-regulated protein was heat shock 70kDa protein 5 and up-regulated proteins were chaperonin and 2-phospho -pyruvate-hydratase ${\alpha}-enolase$ by 1.5mg/ml of CTF-HAS. Discussion : Proteomic analysis approach were performed to screen the differential expression genes. The screened genes will be used for the better understanding in therapeutic effect of CTF-HAS on cancer field.

Genetic Variation of High Molecular Weight Glutenin (HMW-Glu) Subunit in Korean Wheat

  • Hong, Byung-Hee;Park, Chul-Soo
    • 한국작물학회지
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    • 제43권4호
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    • pp.259-263
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    • 1998
  • High molecular weight glutenin (HMW-Glu) subunit compositions of 73 Korean wheat cultivars and experimental lines were evaluated by using one dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis. This method is suitable for obtaining a good resolution of 1Dx2 and 1Ax2$^*$ without adverse effects on separation of other HMW-Glu subunits. Korean wheats examined in this study could be divided into 15 different groups on the basis of HMW-Glu subunit compositions. From the wheat lines tested, it was identified that there were three alleles at the Glu-Al, five at the Glu-Bl and three at the Glu-D1 loci. The null allele of the Glu-Al was occurred in high frequency (79.4%), while low frequencies for 1Ax1 (12.3%) and 1Ax2$^*$(8.2%) were found. High frequency (75.3%) of the subunit pairs of 1Bx7+1By8 at the Glu-Bl loci compared with other subunits was found. The frequencies of subunits 1Dx2. 2+1Dy12 and 1Dx2+1Dy12 from the Glu-D1 loci were 54. 8% and 37.0%, respectively. However, a few Korean wheat lines (8.2%) carried 1Dx5 + 1Dy10 subunit pair which are responsible for good breadmaking quality. The information of HMW-Glu subunit compositions provide a useful tool to characterize wheat lines, and can be directly used in selection of breeding lines of different end-use properties.

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넙치(Paralichthys olivaceus) 혈청단백질의 이차원전기영동분석 조건확립 (Optimization of Conditions for Two-Dimensional Electrophoresis of the Flounder (Paralichthys olivaceus) Serum)

  • 한윤희;남보혜;감영옥;김우진;공희정;이상준;최태진
    • 한국해양바이오학회지
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    • 제2권1호
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    • pp.68-72
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    • 2007
  • 넙치의 혈청 단백질체를 연구하기 위해 본 연구에서는 이차원전기영동 분석 시의 기본조건을 확립하였다. 넙치의 혈청단백질을 이차원전기영동 법으로 분석하기 위해서는 TCA 침전을 이용하여 혈청 내에 고농도로 존재하는 이온물질을 제거하는 전처리과 정이 반드시 요구되었다. 또한 분석 단백질의 농도는 $30{\mu}g$이 적당하였다. 넙치 10개체의 혈청단백질을 이용하여 총 51회의 이차원전기영동을 수행한 결과 1,8207개의 단백질로 이루어진 넙치 혈청단백질 표준 지도를 작성할 수 있었다.

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DGGE에 의한 남태평양 해면 Dactylospongia metachromia의 공생세균 다양성 (Bacterial Diversity of the South Pacific Sponge, Dactylospongia metachromia Based on DGGE Fingerprinting)

  • 정인혜;박진숙
    • 미생물학회지
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    • 제49권4호
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    • pp.377-382
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    • 2013
  • 2012년 2월 미크로네시아의 축(Chuuk)주에서 채집한 해양해면 Dactylospongia meachromia의 공생세균의 주요 군집구조를 Denaturing Gradient Gel Electrophoresis (DGGE) 방법을 이용하여 분석하였다. D. metachromia의 두 개체 CH607과 CH840를 이용하여 DGGE 분석을 수행한 결과, 동종의 두 개체 해면에서 동일한 밴드 패턴을 나타내었다. DGGE 밴드로부터 DNA를 추출하여 염기서열을 분석한 결과, 알려진 염기서열들과 93-100%의 유사도를 나타내었으며 밴드로부터 밝혀진 모든 서열들은 배양되지 않은 세균 클론들과 높은 상동성을 나타내었다. D. metachromia (CH607, CH840)의 공생세균 군집구조는 Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria, Acidobacteria, Actinobacteria, Chloroflexi, Cyanobacteria, Spirochaetes로 총 6문 8강으로 구성되었으며 동일한 지역에서 채집한 같은 종의 해면은 동일한 공생세균 군집구조를 나타냄을 확인하였다.

Association Study between Vitamin D Receptor Gene Polymorphism and Adult Periodontitis in Korea

  • Kang, Byung-Yong;Ha, Nam-Joo
    • Animal cells and systems
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    • 제7권2호
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    • pp.145-149
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    • 2003
  • Adult periodontitis is a chronic inflammatory disease whose etiology is not well defined. Recent studies have shown that vitamin D receptor gene has been a candidate for the susceptibility of adult periodontitis. The purpose of this study is to investigate the frequency of Taq I restriction fragment length polymorphism (RFLP) in the vitamin D receptor gene in nan periodontically healthy controls and 28 adult periodontitis patients. Taq I RFLP in the vitamin D receptor gene was detected by PCR amplification, followed by restriction enzyme digestion and 2% agarose gel electrophoresis. There was no significant difference in the distribution of Taq I RFLP between healthy controls and adult periodontitis group (P > 0.05). Thus, Taq I RFLP in the vitamin D receptor gene may not confer the susceptibility to adult periodontitis in Korean population. However, t allele distributions of this RFLP showed various frequencies among ethnic groups studied. Further studies in other ethnic groups will be required.