• Title/Summary/Keyword: 2D-PAGE

검색결과 323건 처리시간 0.024초

비트 방식 홀로그램 정보저장 장치의 다중화 방법 (A Multiplexing Method using HOE's for Bit-Type Holographic Data Storages)

  • 박우제;김성필;송석호;오차환;김필수;김지덕
    • 한국광학회지
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    • 제16권5호
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    • pp.462-468
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    • 2005
  • 비트 방식 홀로그램 정보저장 장치는 페이지 방식에 비해서 상용화 하는데 있어서 구성이 간단하고 작은 크기로 설계가 유리한 점 등의 많은 장점을 가지고 있다. 하지만 페이지 방식처럼 하나의 기록점에 2차원 면 정보를 이용해서 높은 저장밀도를 기대 할 수 없는 대신 다중화의 다양화가 필요하다. 본 논문에서는 회전 다중화와 각도 다중화를 동시에 용이하게 할 수 있는 비트 방식 홀로그램 정보저장 장치용 광학계를 제안하고 광학계의 선택도 및 특성 분석, 초점면에 72개의 비트 홀로그램 다중화 실험, 기존 DVD의 10배 정도되는 단위면적당 저장밀도 산출로 본 광학계의 가능성을 검증하였다.

Characterization of Alpha Amylase Producing Thielaviopsis ethacetica and Its Raw Starch Hydrolyzing Ability on Different Agricultural Substrates

  • Dissanayaka, Dissanayaka M.S.;De Silva, Sembukuttige N.T.;Attanayaka, D.P.S.T.G.;Kurera, Mihidukulasuriya J.M.S.;Fernando, Charakrawarthige A.N.
    • 한국미생물·생명공학회지
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    • 제47권3호
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    • pp.412-422
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    • 2019
  • The present study reports the morphological and molecular characterization of the fungal strain, CMSS06 and evaluates its raw starch hydrolyzing ability in four different agricultural substrates (rice bran, banana peel, cassava tubers, and coconut water). The potential use of each agricultural substrate to replace the expensive fermentation media was evaluated with six different fermentation media: rice bran (RB), banana peel (BP), cassava starch (CS), cassava in coconut water (CSCW), cassava in modified coconut water (CMCW), and pure Coconut water (CW). The fungal strain CMSS06 was identified as Thielaviopsis ethacetica by the analysis of the ITS sequences. The T. ethacetica alpha amylase enzyme exhibited maximum alpha amylase activity at 72 h, pH 7.0, and $40^{\circ}C$ on soluble starch. This species resulted in the highest enzyme activity (mU/ml) of 26.06, 10.89, 58.82, 14.2, and 54.67 with the RB, BP, CS, CSCW, and CMCW fermentation media, respectively. The results indicate that CS can be used as a carbon substrate and CMCW can be used to accelerate the fermentation by T. ethacetica. The enzyme was partially purified by 40-60% ammonium sulphate fraction, and it showed total enzyme activity, total protein content, specific activity, purification fold, and a recovery of 2400 mU, 30 mg, 80 mU/mg, 2.7, and 71.1%, respectively. The molecular mass of the T. ethacetica alpha amylase was estimated on SDS-PAGE, and two bands around 50 kDa and 70 kDa were identified. The present study implies that T. ethacetica can produce alpha amylase, and it can be used to hydrolyze raw starch during the fermentation processes.

Molecular Cloning, Characterization, and Application of Organic Solvent-Stable and Detergent-Compatible Thermostable Alkaline Protease from Geobacillus thermoglucosidasius SKF4

  • Suleiman D Allison;Nur AdeelaYasid;Fairolniza Mohd Shariff; Nor'Aini Abdul Rahman
    • Journal of Microbiology and Biotechnology
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    • 제34권2호
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    • pp.436-456
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    • 2024
  • Several thermostable proteases have been identified, yet only a handful have undergone the processes of cloning, comprehensive characterization, and full exploitation in various industrial applications. Our primary aim in this study was to clone a thermostable alkaline protease from a thermophilic bacterium and assess its potential for use in various industries. The research involved the amplification of the SpSKF4 protease gene, a thermostable alkaline serine protease obtained from the Geobacillus thermoglucosidasius SKF4 bacterium through polymerase chain reaction (PCR). The purified recombinant SpSKF4 protease was characterized, followed by evaluation of its possible industrial applications. The analysis of the gene sequence revealed an open reading frame (ORF) consisting of 1,206 bp, coding for a protein containing 401 amino acids. The cloned gene was expressed in Escherichia coli. The molecular weight of the enzyme was measured at 28 kDa using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The partially purified enzyme has its highest activity at a pH of 10 and a temperature of 80℃. In addition, the enzyme showed a half-life of 15 h at 80℃, and there was a 60% increase in its activity at 10 mM Ca2+ concentration. The activity of the protease was completely inhibited (100%) by phenylmethylsulfonyl fluoride (PMSF); however, the addition of sodium dodecyl sulfate (SDS) resulted in a 20% increase in activity. The enzyme was also stable in various organic solvents and in certain commercial detergents. Furthermore, the enzyme exhibited strong potential for industrial use, particularly as a detergent additive and for facilitating the recovery of silver from X-ray film.

Identification of Bovine Pregnancy-Specific Whey Proteins using Two-Dimensional Gel Electrophoresis

  • Han, Rong-Xun;Choi, Su-Min;Kim, Myung-Youn;Quan, Yan Shi;Kim, Baek-Chul;Diao, Yun Fei;Koqani, Reza;Park, Chang-Sik;Jin, Dong-Il
    • Reproductive and Developmental Biology
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    • 제32권4호
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    • pp.255-261
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    • 2008
  • The early diagnosis of bovine pregnancy is an essential component of successful reproductive planning on farms, because lack of bovine pregnancy over the long term results in reproductive failure and low milk yield-the latter of which is a special concern on dairy farms. This study was designed to identify early pregnancy-specific whey proteins in bovine, by comparing milk samples collected from cattle during pregnancy (Days 30 and 50) and from non-pregnant cattle. In this study, differentially expressed proteins in five pregnant and five non-pregnant Holstein dairy cattle were investigated and compared, using proteomics analysis. The first dimension was applied to a pH $3.0{\sim}10.0$ strip, by loading a 2-mg milk protein sample. After the second-dimension separation was performed, the gels were stained with colloidal Coomassie brilliant blue. The stained gels were scanned and the images were analyzed, to detect variations in protein spots between non-pregnant and pregnant cattle milk protein spots, using ImageMaster, this was followed by analysis with MALDI TOF-MS. Analysis of the 2-DE gel image resulted in a total of approximately $500{\sim}600$ protein spots, of which 12 spots were differentially expressed, six spots were up-regulated, and four spots were down-regulated; two spots were identified as pregnancy-specific proteins. These proteins were identified as lactoferrin, NA-DH dehydrogenase subunit 2, albumin, serum albumin precursor and transferrin. Our results via 2-D PAGE analysis revealed composite profiles of several milk proteins related to early bovine pregnancy, implying the possible use of these milk proteins in the early detection of bovine pregnancy.

The Protein and Isozyme Patterns During in vitro Plant Regeneration of Yooja (Citrus junos Sieb.) and Trifoliate Orange (Poncirus trifoliata Rafin.)

  • Park, Min-Hee;Jang, Hyun-Kyu;Cha, Young-Ju;Kim, Ho-Bun;Lee, Sook-Young
    • Plant Resources
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    • 제5권1호
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    • pp.29-44
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    • 2002
  • In this study, plant regeneration through in vitro culture from plantlet stems of Yooja (C. junos Sieb.) and trifoliate orange (P. trifoliata Rafin.) was attempted to make mass-production system of virus-free plants having the same genotype with mother plant. In order to investigate physiological change depending on the developmental stage of plant regeneration, the changes of total protein, peroxidase and esterase activity and their isozyme patterns as well were examined in 1/2 MS medium. The results are as follows : 1. The MS medium for the optimal callus induction and shoot formation was utilized. The medium was supplemented either with 2,4-D and Kinetin or with BA and NAA. The optimal concentrations were the combination of 1.0mg/ 2,4-D +0.3mg/ Kinetin and 1.0mg BA +0.3mg NAA in callus induction and shoot formation, respectively. 2. For the plant regeneration from somatic embryos, 1/2 MS medium was used with supplements of growth regulators (free, 1.0mg/ IBA +1.0mg/ BA ,0.5mg/ IBA +0.5mg/ BA). Shooting and rooting were the best in the treatment of 0.5mg/ IBA and 0.5mg/ BA combination. 3. The total protein content has a tendency of increase with the developmental stage of embryo, but it was decreased at the plantlet. Also it was the highest at 8 and 6 weeks stage in C. junos Sieb. and P. trioliata Rafin, respectively. In the SDS-PAGE pattern of protein, C. junos Sieb. showed bands of 29.0 and 40kDa at 10 weeks. The 45,66 and 97.4 kDa bands at 10 weeks of culture were shown in P. trifoliata Rafin. 4. The highest esterase activity was shown at the 6 and 8 weeks of culture in C.junos Sieb. and P. trifoliata Rafin.., respectively. 5. Esterase isozyme patterns were shown difference according to the developmental stage. In C. junos Sieb. a new band was observed at pl 7.7 following 4 weeks culture. On the other hand, new bands in P. trifoliata Rafin. were observed at pl 7.5~6.5 following 4 and 6 weeks culture, respectively.

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넙치 근소포체 및 근원섬유의 생화학적 특성에 미치는 전기자극의 영향 (Effects of Electrical Stimulation on the Biochemical Properties of Plaice, Paralichthys olivaceus, Sarcoplasmic Reticulum and Myofibrils)

  • 김태진;최영준;김동수;조영제
    • 한국수산과학회지
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    • 제31권4호
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    • pp.545-552
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    • 1998
  • 생선회의 육질을 향상시키는 연구의 일환으로 어육에 있어서 전기자극에 의한 근수축의 증대원인을 밝히고자 전기자극처리가 근소포체의 특성에 미치는 영향을 살펴보았다. 넙치 근소포체의 $Ca^{2+}$-ATPase는 $50^{\circ}C$ 이상의 온도에서 실활되었으며, 전기자극시킨 경우 즉살한 것에 비하여 낮은 $Ca^{2+}$-ATPase활성을 나타내었다. 근육을 $5^{\circ}C$에 저장하였을때 시간이 길어질수록 근소포체의 $Ca^{2+}$-ATPase는 치사직후에 비하여 저하되는 경향을 보였으며, 전기자극 시간이 길어질수록 빠르게 저하되었고. 35초와 60초간 전기 자극시킨 것은 비슷한 저하속도를 나타내었다. SDS-PAGE 결과, 97kDa과 68kDa의 성분이 주된 구성단백질이었으며. 전기자극 시킨것은 즉살시킨 것에 비하여 97kDa의 성분이 감소되었고 전기자극 시간이 길어질수록 현저하였다. LSR은 $27\~32\%$ sucrose 농도에서, HSR은 $38\~45\%$의 농도에서 얻을 수 있었다. LSR의 $Ca^{2+}$-ATPase는 전기자극에 의하여 실활되었으며, HSR은 큰 영향을 받지 않았다 근원섬유의 $Mg^{2+}$-ATPase 활성은 전기자극처리에 의하여 증가되었으며 자극시간이 길어질수록 저장 중 $Mg^{2+}(+Ca^{2+}$)-ATPase 활성의 저하는 즉살한 것에 비하여 빠르게 진행되었다. $Mg^{2+}(-Ca^{2+}$)-ATPase 활성의 변화는 $Mg^{2+}(+Ca^{2+}$)-ATPase 활성과 비슷한 경향을 나타내었으며, 전기자극한 것은 저장 중 저하되었으나 즉살한 것은 치사 직후와 비교하여 큰 차이를 나타내지 않았다. 근원섬유의 $Ca^{2+}$-감수성은 즉살과 전기자극한 것 사이에 차이를 나타내지 않았으며, 저장 중에도 변화를 보이지 않았다.

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Butachlor가 귀리의 세포분열(細胞分裂) 및 단백질(蛋白質) 합성(合成)에 미치는 영향(影響) (Effects of Butachlor on the Cell Division and Protein Synthesis on Oat(Avena sativa L.))

  • 권성환;김재철
    • 한국잡초학회지
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    • 제9권3호
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    • pp.245-249
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    • 1989
  • 식물생장(植物生長) 억제제(抑制劑)인 butachlor의 작용기구(作用機構)를 구명(究明)하기 위하여 본(本) 제초제(除草劑)가 생장기본요소(生長基本要素)인 세포분열(細胞分裂) 및 단백질(蛋白質) 합성(合成) 억제(抑制)에 미치는 영향(影響)은 다음과 같다. 1. Butachlor의 세포분열(細胞分裂) 억제(抑制) 효과(效果)는 18시간(時間) 처리후(處理後) $10^{-6}M$은 9.3%, $10^{-5}M$은 28.2%의 억제(抑制)를 보였으며, 특히 $10^{-3}M$ 농도(濃度)에서는 81.6%의 현저한 억제(抑制)를 보임으로서 본(本) 제초제(除草劑)는 농도(濃度)가 높아질수록 세포분열(細胞分裂) 억제효과(抑制效果)도 증가(增加)하였다. 2. 단백질(蛋白質) 억제(抑制) 효과(效果)는 18시간(時間) 처리후(處理後) $10^{-4}M$ 농도(濃度)에서 22.9%, $10^{-3}M$은 34.1%의 억제효과(抑制效果)를 보였다. 또한 24시간(時間) 처리후(處理時) $10^{-4}M$은 34.3%로 로서 18시간(時間) 처리후(處理後)의 $10^{-3}M$과 거의 같은 억제효과(抑制效果)를 보이고 있다. 이와같이 butachlor 는 농도(濃度)와 처리시간(處理時間)이 길어짐에 따라서 단백질(蛋白質) 농도(濃度)의 증가(增加)를 보였다. 3. Butachlor 처리(處理)는 전체적인 단백질(蛋白質) 합성(合成)을 억제(抑制)하였고 특히 분자량 16, 18, 30, 43 그리고 43.5KD의 단백질(蛋白質)을 억제(抑制)하는 것으로 나타났으며 귀리(Avena sativa L.)의 root tip은 100,000 이하의 polypeptide subunit의 단백질로 구성(構成)되었다.

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Soluble Expression of a Human MnSOD and Hirudin Fusion Protein in Escherichia coli, and Its Effects on Metastasis and Invasion of 95-D Cells

  • Yi, Shanze;Niu, Dewei;Bai, Fang;Li, Shuaiguang;Huang, Luyuan;He, Wenyan;Prasad, Anand;Czachor, Alexander;Tan, Lee Charles;Kolliputi, Narasaiah;Wang, Feng
    • Journal of Microbiology and Biotechnology
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    • 제26권11호
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    • pp.1881-1890
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    • 2016
  • Manganese superoxide dismutase (MnSOD) is a vital enzyme that protects cells from free radicals through eliminating superoxide radicals ($O^{2-}$). Hirudin, a kind of small active peptide molecule, is one of the strongest anticoagulants that can effectively cure thrombus diseases. In this study, we fused Hirudin to the C terminus of human MnSOD with the GGGGS linker to generate a novel dual-feature fusion protein, denoted as hMnSOD-Hirudin. The hMnSOD-Hirudin gene fragment was cloned into the pET15b (SmaI, CIAP) vector, forming a recombinant pET15b-hMnSOD-Hirudin plasmid, and then was transferred into Escherichia coli strain Rosetta-gami for expression. SDS-PAGE was used to detect the fusion protein, which was expected to be about 30 kDa upon IPTG induction. Furthermore, the hMnSOD-Hirudin protein was heavily detected as a soluble form in the supernatant. The purification rate observed after Ni NTA affinity chromatography was above 95%. The hMnSOD-Hirudin protein yield reached 67.25 mg per liter of bacterial culture. The identity of the purified protein was confirmed by western blotting. The hMnSOD-Hirudin protein activity assay evinced that the antioxidation activity of the hMnSOD-Hirudin protein obtained was $2,444.0{\pm}96.0U/mg$, and the anticoagulant activity of the hMnSOD-Hirudin protein was $599.0{\pm}35.0ATU/mg$. In addition, in vitro bioactivity assay showed that the hMnSOD-Hirudin protein had no or little cytotoxicity in H9c2, HK-2, and H9 (human $CD_4{^+}$, T cell) cell lines. Transwell migration assay and invasion assay showed that the hMnSOD-Hirudin protein could suppress human lung cancer 95-D cell metastasis and invasion in vitro.

생쥐의 纖維芽細胞와 SCK 腫瘍細胞에서 Stress와 pH에 의한 Stress Protein 遺傳子發見의 調節 (Modulation of Stress Protein Gene Expression by Environmental Stress and pH in the Mouse Fibroblasts and SCK Tumor Cells)

  • Kang, Man-Sik;Lee, Chung-Choo;Lee, Bonggeun;Suh, Mi-Young
    • 한국동물학회지
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    • 제28권2호
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    • pp.108-119
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    • 1985
  • Stress protein (SP) 遺傳子發現의 調節機構를 밝히기 위한 한가지 방편으로 환경의 stress와 pH가 SP의 合成誘導에 어떤 作用을 하는지를 SDS-PAGE를 이용해서 分析하였다. 蛋白質合成의 전반적 양상은 MEF와 SCK 세포에서 달랐으나 SP의 양상은 동일하였다. 그중에서 $SP_70$의 誘導와 感衰의 kinetics는 특히 흥미로웠다. $SP_70$의 kinetics는 酸性 pH와 正常 pH에서 類似하였으나 最大量의 SP 合成에 필요한 溫度와 그 處理時間은 pH에 의해 달리 나타나서, 酸性 pH 에서는 자은 溫度와 짧은 處理時間에서 나타나고 더욱 오래 지속되는 경향을 보였다. SP의 合成誘導와 SP mRNA의 축적은 actinomycin D에 의해 阻止되는 사실로 미루어 SP의 合成이 誘導되기 위해서는 새로운 mRNA의 合成이 필요함을 알수 있었고, cycloheximide 처리의 결과는 SP의 合成誘導에 앞서서 어떤 特異한 蛋白質의 合成은 일어나지는 않음을 알수 있었다. 이상과 같은 몇가지 實驗結果는 MEF와 SCK 세포에서 SP의 合成誘導는 일차적으로 轉寫水潗에서 調節되며, $SP_70$의 合成은 自動調節됨과 아울러 SP의 水潗은 세포늬 stress 상태와 相關關係가 있는 것으로 推論할수 있음을 보여주었다.

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Genetic Variation and Genetic Relationship of Seventeen Chinese Indigenous Pig Breeds Using Ten Serum Protein Loci

  • Mo, D.L.;Liu, B.;Wang, Z.G.;Zhao, S.H.;Yu, M.;Fan, B.;Li, M.H.;Yang, S.L.;Zhang, G.X.;Xiong, T.A.;Li, K.
    • Asian-Australasian Journal of Animal Sciences
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    • 제16권7호
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    • pp.939-945
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    • 2003
  • Seventeen Chinese indigenous pig breeds and three introduced pig breeds had been carried out by means of vertical polyacrylamide gel electrophoresis (PAGE). According to the results, eight serum protein loci were highly polymorphic except Pi-2 and Cp. The polymorphism information content (PIC) of Hpx was the highest (0.5268), while that of Cp was the lowest (0.0257). The population genetic variation index showed that about 84% genetic variation existed in the population, and the rest of 16% distributed between the populations. The genetic variation of Yimeng black pig and Duroc were the highest and the lowest, respectively. The genetic variation of Chinese indigenous pig breeds was much more than that of exotic groups. Genetic distance results showed that Chinese indigenous pig breeds were classified into four groups with the three introduced pig breeds clustered into another group. The results also supported the geographic distribution of Chinese indigenous pig breeds in certain extent.