• 제목/요약/키워드: S-100 protein

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Streptomyces longwoodensis로부터 Autoregulator Receptor Protein 유전자의 클로닝 및 특성 (Characterization and Cloning of the Gene Encoding Autoregulator Receptor Protein from Streptomyces longwoodensis)

  • 여수환;이성봉;김현수
    • 한국미생물·생명공학회지
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    • 제33권2호
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    • pp.96-105
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    • 2005
  • 공시균인 S. longwoodensis IFO 14251의 autoregulators 및 receptor gene 탐색의 일환으로 기존의 Streptomyces속 receptor gene의 공통배열을 primer로 이용하여 PCR을 수행하였다. 예상되는 100 bp 크기의 단편을 pUC19 vector에 ligation하여 E. coli $DH5{\alpha}$에 transformation한 후, plasmid를 분리하여 BamHI을 처리하여 $2\%$ agarose gel에 전기영동한 결과, pUC19 외에 receptor gene PCR product가 100 bp 위치에 존재하는 것을 확인하였다. 형질전환된 plasmid로 PCR을 수행한 후, 염기배열을 결정하여 분석한 결과, Streptomyces sp. 유래의 receptor gene의 일부분임이 확인되었다. 따라서 S. longwoodensis IFO 14251에는 lysocellin 생산에 관여한다고 추정되는 autoregulator receptor protein을 코드하는 유전자가 존재할 것으로 예상되어 100 bp의 PCR product를 probe로 이용하여 Southern 및 colony hybridization을 통하여 4.4 kb의 SphI 단편을 가지는 plasmid(pSLT)를 제작하였고 이를 sequencing한 결과, Streptomyces 속 유래의 autoregulator receptor 단백질을 코드하는 유전자와 3개의 open reading frame(651 bp)을 확인하여 sltR이라 명명하였다. 유전자 해석 결과, 기존의 autoregulator receptor proteins과 비교시 $35{\sim}46\%$의 homology를 나타내었다. 재조합 단백질의 발현을 위해, sltR/pET-l7b plasmid를 제작하고 E. coli BL21(DE3)/pLysS을 host cell로 이용하여 재조합 단백질을 발현시켰다. SltR 재조합 단백질의 정제는 DEAE-Sephacel column chromatography와 DEAE-5PW chromatography(HPLC)를 통해 수행하였다. Gel filtration chromatography(HPLC, 55 kDa)와 SDS-PAGE(28 kDa)를 통해 분자량을 확인한 결과, 재조합 단백질은 dimer형태로 존재하는 것으로 확인되었다. 또한, 결합활성에 있어 A-factor type의 autoregulator에 대해 가장 강한 결합활성을 나타내었다.

The Relationship between Milk Protein Phenotypes and Lactation Traits in Brown Swiss and Canadienne

  • Kim, S.;Ng-Kwai-Hang, K.F.;Hayes, J.F.
    • Asian-Australasian Journal of Animal Sciences
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    • 제11권3호
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    • pp.311-317
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    • 1998
  • A total of 1033 Brown Swiss and 610 Canadienne cows were phenotyped for the genetic variants ${\alpha}_{s1}$-casein, ${\beta}$-casein, ${\kappa}$-casein, ${\beta}$-lactoglobulin and ${\alpha}$-lactalbumin. In Brown Swiss, frequency distributions were: 97.3% B and 2.7% C variant of ${\alpha}_{s1}$-casein; 31.6% $A^1$, 51.8% $A^2$, 0.5% $A^3$ and 16.1% B variant of ${\beta}$-casein; 70.4% A, 29.3% B, and 0.3% C variant of ${\kappa}$-casein; 41.7% A and 58.3% B variant of ${\beta}$-lactoglobulin; and 100% B variant of ${\alpha}$-lactalbumin. Corresponding frequencies in Canadienne for those five milk proteins were: 98.6 and 1.4%;58.5, 33.5, 0.08 and 7.9%; 78.8, 21.1 and 0.1%, 42.4 and 57.6%; and 100%. Analysis of variance by least squares showed possible association between milk protein phenotypes and some lactational production traits. There were no significant association of phenotypes of ${\alpha}_{s1}$-casein, ${\beta}$-casein and ${\beta}$-lactoglobulin with milk yield, fat yield, protein yield, fat percentage and protein percentage in both breeds during the three lactations. In the Brown Swiss, ${\kappa}$-casein phenotype was associated with 305-day fat yield and protein yield during the first lactation. ${\kappa}$-Casein AB was associated with higher milk, fat and protein yield during the second lactation. During the third lactation, ${\beta}$-lactoglobulin AA in Canadienne cows was associated with higher protein content in the milk (3.70%) when compared to phenotypes AB (3.54%) and BB (3.64%).

목질진흙버섯 (Phellinus linteus) HN00K9 균사체 배양 대두의 단백질함량 및 항산화활성 특성 (Characteristics of the protein and antioxidant contents in soybeans cultured with Phellinus linteus HN00K9)

  • 김자윤;백예린;이상호 ;강희완
    • 한국버섯학회지
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    • 제21권3호
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    • pp.126-131
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    • 2023
  • 목질진흙버섯 HN00K9균사체 액체종균을 대두에 배양한 대두(PMS)의 조단백질 함량은 41.99%로 HN00K9 균사체 배양하지 않은 대두(UCS) 38.61%보다 높은 단백질 함량이 검출되었다. PMS와 UCS에서 16종의 유리 아미노산이 검출되었으며 총 아미노산함량은 PMS에서 39,963 mg/100 g으로 UCS의 36,817 mg/100 g보다 8% 이상 증가되었다. 특히 glutamic acid는 PMS에서 7,413 mg/100 g로 전체 아미노산 중 가장 많은 18.5%로 나타났다. 총 polyphenol 함량은 PMS에서 2.66 mg GAE/g로 UCS 1.45 mg GAE/g보다 45%이상 높게 검출되었으며 PMS는 33.3%의 DPPH radical 소거 능을 보여 UCS의 11.5%보다 3배 증가한 높은 항산화 활성을 보였다. 따라서 본 연구의 PMS는 높은 단백질함량과 항산화 활성이 강화된 기능성 식품소재로 활용할 수 있을 것으로 기대된다.

국산 조제분유의 열처리 등급에 관한 기초 연구 (The Fundamental Studies of Heat Treatment Class in Domestic Infant Formula)

  • 박영희
    • 한국식품영양과학회지
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    • 제27권3호
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    • pp.495-500
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    • 1998
  • To compare the extent of heat treatment in domestic infant formula, pH,titratable acidity, undenatured whey protein contents, HMF contents and protein-reducing substances of three commercial products (A, B, C) were measured. The pH of B products was lowest and the titratable acidity of B product was highest. The contents of undenatured whey protein per 100ml serum were 0∼30mg(A products), 90∼130mg(B products)and 80∼90mg(C products), respectively. Distinct differences of undenatured whey protein contents according to the manufacturer and infat's stage in age could be observed. The HMF contents of tested products showed 10.9∼21.5umol/L and B-2 product(B products for the second stage of 5∼9 month) was the highest among tested products. The protein-reducing substances showed 4.46∼9.50mg K4Fe(CN)6/100ml serum nd B-2 product was the highest among tested products.

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사과 겹무늬썩음병균(Botryosphaeria dothidea)에 의해 부패된 사과 과실에서 Pectin질 분해효소의 생산과 Pectin질의 변화 (Production of Pectolytic Enzymes and Change of Pectic Substances from Apple Fruits Infected with Botryosphaeria dothidea)

  • 박석희;이창은
    • 한국균학회지
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    • 제21권2호
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    • pp.106-111
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    • 1993
  • 사과 겹무늬썩음병균 Botryosphaeria dothidea에 의해 부패된 사과과실에서 pectin질 분해효소를 추출하여 그들의 활성과 pectin 성분의 변화를 조사하였다. 본 병원균은 exo-polygalacturonase(exo-PG), exo-polymethylgalacturonase(exo-PMG), polygalacturonate-trans -e1iminase(PGTE)와 pectinmethyl-trans-eliminase(PMTE)를 생산하였다. 부패된 사과 과육에서 exo-PG와 exo-PMG는 접종 후 7일째 specific activity가 각각 21.15 및 24.65 units/mg protein으로 높게 나타났다. PGTE와 PMTE의 활성은 7일째 각각 5.60과 7.90 uints/mg protein으로 나타났으나 exo-type의 효소보다는 그 활성이 낮았다. 수용성 pectin은 부패가 진행됨에 따라 14일째에 11.50 mg/100 mg-AIS이었고, versene-soluble pectin 역시 7.31 mg/100 mg-AIS로 나타나 건전과와 비교하여 각각 4.23 및 2.16 mg/100 mg-AIS 증가하였다. 부패과의 총 가용성 펙틴 함량은 총 pectin의 72.4%로서 건전과와 비교하여 24.8% 더 높았다. 불용성 pectin 함량은 부패가 진행됨에 따라 15.32 mg/100 mg-AIS에서 7.16 mg/100 mg-AIS로 현저하게 감소하였으며, 총 pectin 함량은 큰 변화가 없었다.

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닭 태자의 근육위와 유문부에 대한 조직학적 및 면역조직화학적 연구 (Histological and immunohistochemical studies on the gizzard and pylorus regions of the chicken embryos)

  • 이재현;구세광;이형식
    • 대한수의학회지
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    • 제38권4호
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    • pp.686-695
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    • 1998
  • Histological changes, distributions and relative frequencies of bovine Sp-1/chromogranin (bCG)-, serotonin-, gastrin-, cholecystokinin-8(CCK-8)-, somatostatin-, S-100 protein-, polypeptide YY(PYY)- and glucagon-immunoreactive cells were investigated in the gizzard and pylorus of the chicken embryos from 10 days of incubation to hatching. Histologically, the pseudostratified columnar epithelium were observed from 10 days of incubation to 15 days of incubation, thereafter these epithelium were differentiated to simple columnar epithelium, gastric gland and/or mucosal gland. In the gizzard, bCG-immunoreactive cells were observed from 19 days of incubation and S-100 protein-immunoreactive cells were detected from 15 days of incubation to 18 days of incubation. No serotonin-, gastrin-, CCK-8-, somatostatin-, PYY- and glucagon-immunoreactive cells were found in this region. In the pylorus, bCG-, gastrin- and somatostatin-immunoreactive cells were observed from 16 days of incubation respectively, thereafter these cells were increased with ages. CCK-8-immunoreactive cells were detected on hatching and S-100 protein-immunoreactive cells were detected from 16 days of incubation to 18 days of incubation. No serotonin-, PYY- and glucagon-immunoreactive cells were observed in this region.

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Polyamine에 의한 옥수수 Ribosome의 활성 촉진 (Stimulation of Ribosome Activity of Zea mays by Polyamine)

  • 김기남
    • Journal of Plant Biology
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    • 제36권1호
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    • pp.83-90
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    • 1993
  • As a part of the study on the relation between exogenous polyamines and various components necessary for protein biosynthesis in the germinating maize seeds, the effects of the polyamines on protein biosynthesis and irbosome activity were investigated. The protein biosynthesis activity by S-30 containing all components necessary for protein biosynthesis was increased by exogenous polyamines, spermidine, spermine and putrescine. As the concentration of polyamine treated was increased, the optimal Mg2+ concentration of in vitro poly U-dependent protein synthesis system was gradually reduced. However, the optimal Mg2+ concentration of poly U-dependent system containing optimal polyamine was 10 mM regardless of the sort of polyamine. It could be infered that polyamines play an important part in protein biosynthesis in the higher plant, and that the role of polyamines take partially the place of Mg2+ action. The activities of ribosome and S-100 in protein biosynthesis were increased by 46.7% and 17.7% with spermidine, and by 44.1% and 16.2% with spermine, and by 29.1% and 19.3% with putrescine. It could be concluded that the increase of protein biosynthesis by polyamines in mainly owing to the ribosome activation.

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A Growth-Stimulating Protein in Cow's Milk

  • Ki, Eun-Kyung;Park, Yong-Bok;Choi, Myung-Sook
    • 동아시아식생활학회지
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    • 제4권3호
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    • pp.59-65
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    • 1994
  • 이 실험에서는 소의 유즙에 있는 성장인자를 세포배양을 통해 확인하고 크로마토그래피를 사용하여 분리 정제하였다. 5%의 우유를 포함하는 배지를 FBS(fetal bovine serum)를 포함한 배지와 비교했을때 Africal green monkey kidney cell의 성장 촉진 효과가 비슷하게 나타났으며, 우유의 성장인자를 분리 정제한 결과 FBS배지에서보다 2,000배 이상의 성장 촉진 효과가 나타났다. 이 성장인자는 hydrophobic column(phenyl-sepharose)과 gel-filtration column(sephadex G-100)을 사용하여 분리했으며 그 결과 milk-deriven growth factor는 phenyl-sepharose에서 50% ethylene glycol step에서 용출되므로 hydrophobic protein임이 증명되었고 size exclusion column chromatography로 부터 분자량이 100,000-15,0000 범위의 고분자 물질임이 밝혀졌다. 또한 우유의 성장인자는 매우 다양하며 그중 본 실험으로부터 정제된 물질이 주요 성장인자로 밝혀졌다.

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S100ß, Matrix Metalloproteinase-9, D-dimer, and Heat Shock Protein 70 Are Serologic Biomarkers of Acute Cerebral Infarction in a Mouse Model of Transient MCA Occlusion

  • Choi, Jong-Il;Ha, Sung-Kon;Lim, Dong-Jun;Kim, Sang-Dae;Kim, Se-Hoon
    • Journal of Korean Neurosurgical Society
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    • 제61권5호
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    • pp.548-558
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    • 2018
  • Objective : Diagnosing acute cerebral infarction is crucial in determining prognosis of stroke patients. Although many serologic tests for prompt diagnosis are available, the clinical application of serologic tests is currently limited. We investigated whether $S100{\beta}$, matrix metalloproteinase-9 (MMP-9), D-dimer, and heat shock protein 70 (HSP70) can be used as biomarkers for acute cerebral infarction. Methods : Focal cerebral ischemia was induced using the modified intraluminal filament technique. Mice were randomly assigned to 30-minute occlusion (n=10), 60-minute occlusion (n=10), or sham (n=5) groups. Four hours later, neurological deficits were evaluated and blood samples were obtained. Infarction volumes were calculated and plasma $S100{\beta}$, MMP-9, D-dimer, and HSP70 levels were measured using enzyme-linked immunosorbent assay. Results : The average infarction volume was $12.32{\pm}2.31mm^3$ and $46.9{\pm}7.43mm^3$ in the 30- and 60-minute groups, respectively. The mean neurological score in the two ischemic groups was $1.6{\pm}0.55$ and $3.2{\pm}0.70$, respectively. $S100{\beta}$, MMP-9, and HSP70 expressions significantly increased after 4 hours of ischemia (p=0.001). Furthermore, $S100{\beta}$ and MMP-9 expressions correlated with infarction volumes (p<0.001) and neurological deficits (p<0.001). There was no significant difference in D-dimer expression between groups (p=0.843). The area under the receiver operating characteristic curve (AUC) showed high sensitivity and specificity for MMP-9, HSP70 (AUC=1), and $S100{\beta}$ (AUC=0.98). Conclusion : $S100{\beta}$, MMP-9, and HSP70 can complement current diagnostic tools to assess cerebral infarction, suggesting their use as potential biomarkers for acute cerebral infarction.