• Title/Summary/Keyword: 단백질 추출

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Agronomic Characteristics and Aromatic Compositions of Korean Wild Codonopsis lanceolata Collections Cultivated in Field (한국 야생더덕 수집종의 노지 재배시 생육 특성과 향기성분 조성)

  • 이승필;김상국;민기군;조지형;최부술;이상철;김길웅
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.41 no.2
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    • pp.188-199
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    • 1996
  • The native ecological environment and aromatic constituents of Korean wild Codonopsis lanceolata and one Japanese strain were investigated to find Codonopsis lanceolata strains showing high aromatics, and to know regional differences among these strains. The results were as follows : There were no remarkable differences among the Korean wild C. lanceolata strains in ecological environments. Recovery yield of essential oils was highest in Togyusan strain with 0.009%. Difference in protein band patterns among these strains was not recognized, and peroxidase and esterase pattern changes were appeared in different collected regions at the leaf and root tissues. Major aromatic constituents were 11 kinds of aliphatic alcohols such as trans-2-hexenal, 1-hexanol, cis-3-hexanol, and trans-2-hexanol. And Togyusan strain, Sobaeksan strain, and Kayasan strain have the highest aliphatic alcohols of plant essential oils. In particular, BHT(butylated hydroxytoluene), one of the antioxidants, was detected in Chirisan strains.

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Isolation of Polyphenol Compounds from the Leaves of Korean Persimmon (Diospyrus kaki L. Folium) (한국산 감잎로부터 Polyphenol계 생리활성물질 분리)

  • An, Bong-Jeun;Bae, Man-Jong;Choi, Hee-Jin;Zhang, Yun-Bin;Sung, Tae-Soo;Choi, Cheong
    • Applied Biological Chemistry
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    • v.45 no.4
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    • pp.212-217
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    • 2002
  • We purified polyphenols from persimmon leaf and tested their biological activity. The 60% acetone extract was lyophilized and applied to test enzyme inhibition of glucosyltransferase and tyrosinase. GTase was 82.4% inhibited at $1.8{\times}10^{-1}$ mg/ml and tyrosinase 21.7% inhibited at 0.8 mg/ml. The acetone extract was fractionated into F-1, 2, 3, 4, 5 by Sephadex Q-50 gel filtration and the fraction-1 and 2 showed higher enzyme inhibition activity than the other fractions. To the Proteinase K treatment and autoclaving of the two fractions had no effect on the enzyme activity, but these results suggested that active fraction was not protein but phenol ring completed compounds. By Sephadex LH-20, MCI-gel and Bondapak $C_{18}$ column chromatographies, compouds 1, 2, 3 and 4 from F-1 fraction, compounds 5 and 6 from F-2 fraction and compounds 7 , 8 from F-3 fraction were purified and re-crystallized. The purified compounds was assumed to be condensed tannins of frame flavan-3-ol frame on the basis of color reagent reaction and to be a mixture of monomer, dimer and trimer according to TLC analysis.

Effect on the Production of Toxin Protein of Bacillus thuringiesis by Culture Conditions (배양 조건이 Bacillus thuringiesis 독소단백질의 생산에 미치는 영향)

  • Kim, Tae Wan;Kim, Tae Hwan;Yu, Yong Man
    • The Korean Journal of Pesticide Science
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    • v.18 no.4
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    • pp.358-363
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    • 2014
  • We was investigated parasporal inclusion proteins change to use industrial medium of new strain Bacillus thuringiensis CAB 565, CAB 566. To confirm medium's oxygen efficient consist of glucose and yeast extract, we was conducted oxygen transfer coefficients (KLa) of medium's concentration and impeller in 20 l-Jar fermentor. When to increase air flow rate and medium concentration, KLa rate is rise. Also it is effective on agitation rate 200 rpm, but KLa rate is decrease when to rise agitation rate. To hold dissolved oxygen rate (upper 50%), Air flow rate is steadily increase on culture to use microsparger. When 16 hour of culture stage, B.t. CAB 565 and B.t. CAB 566 harvested respectively $2.3{\times}10^{10}$, $1.8{\times}10^{10}$ viable cell/ml. When 54 hour, B.t. CAB565, 566 harvested respectively $1.9{\times}10^{10}$, $1.4{\times}10^{10}spore/ml$. To resulting carbon's concentration, It is the most effective that glucose concentration is contained 5% in medium.

Analysis of the Effects of Bone Marrow Biopsy Decalcification Methods on Histopathological Examination (골수생검조직의 조직병리검사에서 탈회방법에 따른 결과 분석)

  • Park, Ji Young;Han, Kyung Hee
    • Korean Journal of Clinical Laboratory Science
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    • v.48 no.4
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    • pp.371-377
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    • 2016
  • Decalcification is routinely performed to obtain a pathological diagnosis using bone marrow biopsy. During the decalcification process using a conventional acidic solution, such as HCl, the antigenicity of tissue is damaged. Especially DNA and RNA in the bone marrow are impaired. Hence, there is the need for a standardized decalcification protocol that preserves the antigenicity of tissue. To this end, we compared the effects of two commonly used decalcifiers: Commercial decalcifier (Calcl-Clear Rapid, HCl) and the EDTA (12.5%, pH 7.0). Bone marrow biopsies sampled from 71 patients were decalcified in accordance with the protocols of respective groups-HCI versus EDTA. The differences of decalcification protocols were analyzed with respect to Hematoxylin & Eosin staining, Gomori'sreticulum staining, and immunohistochemical staining and molecular analysis. Immunohistochemical staining used Ki-67, CD20 and CD138 as primary antibodies and molecular analysis was conducted through the DNA concentration analysis, in situ hybridization (ISH) and immunoglobulin heavy chain (IGH) gene rearrangement. On the routine histopathology analysis, there was no difference between HCl and EDTA. Moreover, in case of immunohistochemical staining, the cytoplasmic membrane or cytoplasmic CD markers was well preserved. However, nuclear proteins, such as Ki-67, were stained with low quality. Conversely, according to the molecular analysis, the EDTA protocol preserved the DNA and RNA compared with the HCI. The differences of DNA quantity and quality were statistically significant between protocols of HCl and EDTA. We used 38 cases in HCl and 12 cases in EDTA. Consequently, the EDTA protocol maintains the antigenicity of the protein on tissue and is acceptable for examination with molecular base analysis. Decalcification of bone marrow biopsy by EDTA is highly recommended for the examination of immunohistochemical staining and molecular analysis.

Silymarin Attenuates Invasion and Migration through the Regulation of Epithelial-mesenchymal Transition in Huh7 Cells (간암세포주에서 상피간엽전환억제를 통한 Silymarin의 침윤 및 전이 억제 효과)

  • Kim, Do-Hoon;Park, So-Jeong;Lee, Seung-Yeon;Yoon, Hyun-Seo;Park, Chung Mu
    • Korean Journal of Clinical Laboratory Science
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    • v.50 no.3
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    • pp.337-344
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    • 2018
  • Hepatocellular carcinoma (HCC), a major type of hepatoma, is associated with high recurrence and mortality because of its uncontrolled metastatic feature. Silymarin is a polyphenolic flavonoid from Silybum marianun (milk thistle) and exhibits anti-carcinogenic activity through modulation of the epithelial-mesenchymal transition (EMT) in several cancer cells. In this study, the inhibitory mechanism of silymarin against migration and invasion was investigated in the Huh7 HCC cell line. Wound healing and in vitro invasion assays were conducted to examine the effects of silymarin on migration and invasion. Western blot analysis was also applied to evaluate the inhibitory effects of silymarin on the EMT-related genes and their upstream signaling molecules. Silymarin inhibited the migratory and invasive activities of Huh7 cells. In addition, silymarin attenuated the protein expression levels of vimentin and matrix metalloproteinase (MMP)-9 as well as their transcription factors, Snail, and nuclear factor $(NF)-{\kappa}B$, while the expression of E-cadherin was increased by the silymarin treatment. Among the upstream signaling molecules, the phosphorylation of Akt was inhibited by the silymarin treatment, which was confirmed by the selective inhibitor, LY294002. Consequently, silymarin inhibited the invasive and migratory activities in Huh7 cells through the modulation of EMT-related gene expression by the PI3K/Akt signaling pathway, which may have potential as a chemopreventive agent against HCC metastasis.

Inhibitory Effects and Molecular Mechanism of Adipocyte Differentiation by Rosae laevigata Fructus Ethanol Extracs (금앵자 에탄올 추출물에 의한 3T3-L1 지방세포의 분화억제 효과와 그 메커니즘 규명)

  • Jeong, Hyun Young;Jeong, In Kyo;Nam, So Yeon;Yun, Hee Jung;Kim, Byung Woo;Kwon, Hyun Ju
    • Microbiology and Biotechnology Letters
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    • v.44 no.1
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    • pp.89-97
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    • 2016
  • Obesity is caused by excess accumulation of body fat and contributes to various pathological disorders such as diabetes, hypertension, cardiovascular disease, and cancer. In this study, we investigated the effect of a 30% ethanol extract of Fructus Rosae laevigata (RLE) on adipogenesis in 3T3-L1 adipocytes, measured by triglyceride accumulation and expression of adipogenesis-related transcription factors during differentiation of pre-adipocytes into adipocytes. RLE decreased the intracellular triglyceride contents (assessed by Oil Red-O staining) in a dose-dependent manner. It also downregulated the expression of adipogenic transcription factors and inhibited cell proliferation during the mitotic clonal expansion phase of adipocyte differentiation by inducing G1 phase arrest. We investigated the alterations in the levels of G1 phase arrest-related proteins. The expression of p21 protein significantly increased, while the levels of Cyclin E, Cdk2, and phospho-Rb decreased in a dose-dependent manner in 3T3-L1 cells treated with RLE. These results suggest that RLE inhibits the differentiation of 3T3-L1 adipocytes by suppressing the expression of adipogenic transcription factors and inducing G1 phase arrest in the early stages of adipocyte differentiation.

Study on Identification and Purification of Germanium-fortified Yeast (게르마늄강화효모의 게르마늄결합 단백질의 분리 및 확인에 관한 연구)

  • Lee, Sung-Hee;Lee, Sang-Kwang;Lee, Hyun-Joo;Yi, Yong-Sub;Park, Eun-Woo
    • Applied Biological Chemistry
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    • v.49 no.1
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    • pp.55-59
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    • 2006
  • This study was designed to investigate the optimum manufacturing condition of germanium-fortified yeast, and the binding properties of germanium (Ge) in germanium-fortified yeast. The nutritional optimum conditions were glucose 3.0 (w/v) %, yeast extracts 0.3 (w/v) % and peptone 0.5 (w/v) %, and the amounts of yeast cells were 67.4 mg/ml. And, the standard germanium-fortified yeast was produced under the condition at the ratio of yeast cell and germanium solution was 1 : 0.5 (50%), pH 6.5 and $35-40^{\circ}C$ during fermentation. In results of the identification, binding of germanium-protein showed structural difference between the inorganic Ge $(GeO_2)$ added during fermentation process and germanium-fortified yeast. Therefore, germanium-fortified yeast made by biosynthetic technology formed structurally safe organic germanium during fermentation process. Germanium-fortified yeast can be applied as a new functional material far the improvement of health, the prevention and treatment of chronic degenerative disease like cancer, and the enforcement of immune system.

섬유소 분해효소 유전자 도입 형질전환 돼지 생산

  • 이연근;박진기;민관식;성환후;임기순;양병철;김진회;류재웅;장원경
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.58-58
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    • 2001
  • 본 연구는 섬유소 분해효소 유전자 Cel D(Cellulase Digestion)가 도입된 형질전환 돼지 생산을 통하여 섬유소 함유 사료의 이용 효율을 증대시키고, 나아가 장기이식동물 및 고가의 의료용 단백질 생산가축을 개발하기 위한 원천기술 확보에 있다. 섬유소분해 유전자의 크로닝 및 조직 특이적 발현벡터를 개발하기 위하여, 우선 소의 위중 제4위 내의 미생물로부터 전체 유전자를 분리하였고, 이렇게 작성된 DNA library에서 섬유소분해 관련 유전자인 약 2.0 kb의 Cel D유전자를 크로닝하였으며, 췌장 특이적 발현 프로모터(rat elastase I: 약 200bp)를 크로닝한 후, 미세주입용 형질전환 재조합 벡터를 구축하기 위하여 rElastase I 프로모터 하류에 섬유소 분해 유전자(Cel D)를 연결하여 약 3.0 kb 크기의 재조합 벡터를 준비하였으며, 재조합 유전자를 1세포기 수정란 전핵내에 미세주입 하기 위해 Sal I과 BglII를 이용 유전자 단편을 만들었다. 구축된 유전자를 미세주입하기 위한 수정란을 회수하기위해 총68두의 돼지를 4-5두씩 분리사육하면서 발정동기화 및 과배란 유기를 위해 PG600, Altrenogest, FSH, hCG를 투여하였으며 hCG투여후 약54시간에 외과적 방법에 의해 총 1,359개의 수정란을 회수하였고, 이중 미세주입가능한 1세포기 수정란은 1,296개로 두당 평균 15.9개 였다. 1,296개의 1세포기 수정란 중에서 재조합 유전자(rE I-CelD)가 미세주입된 660개의 수정란을 32두의 수란돈에 외과적 방법에 의해 이식하였으며, 이식되어진 모돈 13두가 분만하여 40.6%의 임신율을 나타내었다. 이렇게 분만된 13두에서 총 65두(암:33두, 수:32두)의 자돈이 생산되었으며, 형질전환 여부를 판명하기 위해 자돈의 꼬리조직으로 부터 genomic DNA를 추출하고 PCR 검정을 실시하였다. PCR 검정 결과, 섬유소 분해 유전자가 도입된 자돈은 5두 이었으며, 그 결과를 Table에 나타내었다.(Table Omitted) Table 1 에서와 같이 섬유소 분해효소유전자가 형질전환된 자돈은 65마리 중 5마리로 7.69%의 형질전환율을 나타내었으며, 5마리의 자돈중 2두(암:1두, 수:1두)는 분만 후 즉시 폐사되었으며 2두(암:1두, 수:1두)는 86일령 그리고 14일령에 폐사하여 현재 1두(암)가 생존하여 섬유소 분해 사양 실험 중에 있다.

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Anti-inflammatory and antimicrobial effects of Jeju rosemary essential oil against skin flora (제주산 로즈마리 에센셜 오일의 항염 및 피부 상재균에 대한 항균 활성)

  • Kim, So-Hee;Yi, Mi-Ran;Kim, Chang-Soo;Kim, Jung-Mi;Bu, Hee-Jung
    • Journal of the Korean Applied Science and Technology
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    • v.35 no.3
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    • pp.744-756
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    • 2018
  • The purpose of this study was to investigate the anti-inflammatory effect and the antimicrobial activity to skin flora of essential oil from rosemary that naturally grown in Jeju. rosemary essential oil was extracted by water distillation essential oil extraction method. In order to confirm the anti-inflammatory activity of rosemary essential oil, it was confirmed that the production of NO and $PGE_2$ induced by LPS in RAW 264.7 cells was inhibited in a concentration-dependent manner. Western blot analysis showed that the expression of iNOS and COX-2, which are biosynthetic enzymes, decreased in a concentration-dependent manner. In addition, production of $TNF-{\alpha}$ and IL-6 the pro-inflammatory cytokines were inhibited. Antimicrobial activities of three S. epidermidis and three P. acnes strains including two antibiotic resistant strains were observed in paper disc method and MIC and MBC tests showed inhibition of bacterial growth and death. From the results of the experiment, we confirmed that rosemary essential oil has the anti-inflammatory and antimicrobial efficacy and it could be used as a cosmetic and skin care material in the future.

A Pilot Study for Development of the Serious Game Contents for Education in the Elderly Diabetes (노인 당뇨환자 교육용 기능성 게임 콘텐츠 개발을 위한 예비연구)

  • Kim, Yu Jeong
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.18 no.7
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    • pp.184-192
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    • 2017
  • This study was a pilot test of serious educational game content(named Roly Poly 160) designed to enhance self-care by elderly diabetics. Roly Poly 160 was developed in eight steps (literature review, demand survey and consultation, extraction of serious content using games, development of Roly Poly 160, pretest for users, workshop for health care workers, final completion of Roly Poly 160 and user satisfaction survey of Roly Poly 160). Roly Poly 160 is intuitively structured in three modules (self-management, card games, and quiz games) that can be self-managed by the elderly and is designed based on six principles. First, we constructed an intuitive interface considering the age of users. Second, the menu was selected as the main menu for Koreans and calorie learning was made at the same time. Third, the calories and nutrients (carbohydrate, fat, protein, calcium, sodium) of selected foods are analyzed, and all test data recorded in the questionnaire chart are graphically displayed by year, month and week so that the change trends can be grasped at a glance. Fourth, necessary data were saved and output and used as educational data. Fifth, user data are made compatible and aggregated and up to 100 million members can be registered. Sixth, it is designed to be developed as a mobile app if necessary. Using Roly Poly 160, 119 diabetic patients were diagnosed with diabetes, and the satisfaction score was 4.26 out of 5. These findings indicate that Roly Poly 160 is appropriate as a diabetes self-care tool and suggests that there is a possibility to use it as a program to educate people about diabetes in public health centers, hospitals and clinics.