• Title/Summary/Keyword: 단백질 특성 분석

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DNA Binding Specificity of Proteus mirabilis Transcription Regulator (Proteus mirabilis 전사 조절 단백질의 DNA 결합 특성)

  • Gang, Jong-Back
    • Korean Journal of Microbiology
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    • v.47 no.2
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    • pp.158-162
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    • 2011
  • Amino acid sequence alignment shows that $\underline{P}$roteus $\underline{m}$irabilis $\underline{t}$ranscription $\underline{r}$egulator (PMTR) has cystein sequence homology at metal binding domain to CueR (copper resistance) protein, which conserves two cysteins (Cys 112 and Cys 120 in PMTR). Gel shift assay revealed that PMTR protein bound to promoter region of Escherichia coli copA (copper-translocating P-type ATPase) and Proteus mirabilis atpase (putative copper-translocating P-type ATPase) genes except that of E. coli zntA (zinc-translocating P-type ATPase) gene. DNase I protection experiment indicated that PMTR protein protected the region over -35 box and close to -10 box. DNase I hypersensitive bases were shown at C and A bases of labeled template strand and at G and C bases of labeled non-template strand of DNA. These hypersensitive bases were appeared in other metalloregulatory proteins of MerR family, which suggests protein-induced DNA bending.

Proteins and Amino Acid Composition of Korea Ginseng Classified by Years (한국인삼의 연근별 단백질 및 아미노산 조성)

  • Choi, Cheong;Yoon, Sang-Hong;Bae, Man-Jong;An, Bong-Jean
    • Korean Journal of Food Science and Technology
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    • v.17 no.1
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    • pp.1-4
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    • 1985
  • For the systematic investigation of biochemical characteristics of Korean ginseng protein by years, protein fractions were analyzed by the techniques of polyacrylamide gel electrophoresis, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and gel filtration, while the amino acid composition was studied by amino acid autoanalyzer. Results of polyacrylamide gel electrophoresis and SDS-PAGE showed a few difference in pattern and number of bands depending on the age of the root. However, the number of bands obtained from polyacrylamide gel electrophoresis and SDS-PAGE was 8 and 7 to 11, respectively. When water extracted proteins were fractionated by Sephadex G-200, the main peak among 2 peaks was collected and lyophillized. Its mol. wt. was extimated to be 43,000 by the SDS-PAGE method. In amino acid composition of water extracted protein and main fraction of gel filtration, arginine content was the highest, 47.17% in water extracted protein and 57.36% in main fraction followed by glutamic acid and asparatic acid. On the contrary, cystine and methionine contents were very low in both cases.

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Production and Characterization of Selenium Peptide from Saccharomyces Cerevisiae (효모를 이용한 selenium peptide 생산 및 특성 연구)

  • 김은기;김영옥;이정옥;이백석
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.30 no.1
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    • pp.73-77
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    • 2004
  • Selenium containing peptide was produced by culturing yeast with selenium, Selenium was broadly incorporated in the various size of proteins based on the GPC analysis of the total yeast protein. The ratio of selenium to protein increased with the concentration of added selenium in the culture medium. Antioxidant activity (glutathione peroxidase-like activity) was proportional to the concentration of selenium concentration in the peptide. Different size of proteins were obtained by hydrolyzing the total yeast protein by protease XIV. Average molecular weight of selenium peptide was analyzed by GPC. Glutathione peroxidase (GPx) activity of the selenium peptide increased as the size of peptide decreased. Sodium selenite had strong inhibition on the yeast growth than sodium selenate. The ratio of selenium to protein was higher with sodium selenate than with sodium selenite. These results showed the potentials of selenium peptide production by yeast cultivation.

Characterization of Recombinant Baculovirus Ewpressing Polyhedrin Gene of Bombyx mori and Autographa californica Nuclear Polyhedrois Virus (두 종류의 다각체 단백질 유전자를 발현하는 유전자 재조합 핵다각체병 바이러스의 특성)

  • 김우진;우수동
    • Journal of Sericultural and Entomological Science
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    • v.38 no.1
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    • pp.36-41
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    • 1996
  • Recombinant baculoviruses having expanded host range were selected by coinfection of Autographa california NPV and Bombyx mori NPV into Sf-9 and BmN-4 insect cell lines. In order to determine the polyhedra morhplogy of RecS-A6, one of a recombinant baculovirus, polyhedra of RecS-A6 produced in insect cells were observed by phase contrast microscope and scanning electron microscope. The results revealed that the recombinant baculovirus had a various polyhedra morphology which was different from its parental viruses, suggesting that the various morhpology of recombinant baculovirus with an expanded host range was due to the genetic recombination of viral genome. To analyze the genomic recombinantion of the recombinant baculoviruses, genomic DNAs of two parent viruses and RecS-A6 were digested with restriction endonuclease and subjected to agarose gel electrophoresis. Southern blot analysis revealed that RecS-A6 has two polyhedrin gene of AcNPV and BmNPV in a viral genome. Polyhedral protein of recombinant baculovirus was analysed by SDS-PAGE. The result showed that molecular weight of polyhedral protein of RecS-A6 containing two polyhedrin gene of AcNPV and BmNPV was as the 31 kDa band of AcNPV and 30 kDa band of BmNPV.

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Construction of a Transgenic Plant to Develop a New Method for the Isolation of Calmodulin-Binding Proteins (새로운 방법을 이용한 칼모둘린 결합 단백질 분리를 위한 형질 전환 식물체의 구축)

  • Kim, Sun-Ho;Lee, Kyung-Hee;Kim, Kyung-Eun;Jung, Mi-Soon;Lim, Chae-Oh;Lee, Shin-Woo;Chung, Woo-Sik
    • Journal of Life Science
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    • v.17 no.9 s.89
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    • pp.1177-1181
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    • 2007
  • Calmodulin (CaM), a ubiquitous calcium-binding protein, regulates diverse cellular functions by modulating the activity of a variety CaM-binding proteins (CaMBPs). Because eukaryotes have multiple CaMBPs, it is important to isolate and characterize them in different tissues and conditions. So far a number of CaMBPs have been identified through classical screening methods. Many classes of proteins have been predicted to bind CaMs based on their structural homology with already known targets. In an effort to develop a method for large-scale analysis of CaMBPs in Arabidopsis, we have generated a transgenic plants overexpressing AtCaM2-GFP. We performed protein pull-down assay to test whether exogenously expressed AtCaM2-GFP proteins can interact with CaMBPs. The exogenously expressed AtCaM2-GFP could strongly interact with a CaMBP, AS1 protein. This result suggests that AtCaM2-GFP in transgenic plants may interact with many CaMBPs in plant cell. Therefore, we will be able to isolate kinds of CaMBPs by using these transgenic plants in many different tissue and environments.

Varietal Difference of Protein Content and Amino Acid Composition in Peanuts (땅콩품종의 단백질 함량과 아미노산 조성)

  • Lee, Jung-Il;Park, Hee-Woon;Kang, Kwang-Hee;Kim, Ki-Joon
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.35 no.5
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    • pp.424-439
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    • 1990
  • The protein content of seed meal were examined to evaluate genotypes of higher protein content from 164 peanut germplasms. The variations in amino acids content were investigated from nine Korean leading varieties grown at five locations. Amino acids contents of protein fractions were also ana lysed in the present study, too. The results of the study were summarized as fallows. The seed protein of 164 peanut varieties averaged 24.5% ranging from 19.5% to 28.7%, showed 9.2% varietal variation in proteins. Differences were not observed in protein content between plant types, however, protein contents were higher in small-seeded than large seeded varieties. The differences were more greater particulaly in Spanish type varieties. The varieties introduced from Taiwan and Philippine showed higher protein contents, and the cultivars or lines bred in Korea and introduced from Japan were lower in protein contents. protein contents showed non-significantly negative correlations with 100-kernel weight and pod weight per plant, but positive correlations were observed between oil and protein content in all types of peanuts. Significant differences among the varieties and locations were observed for total, essential and non-essential amino acids contents, and aspartic acid. Locational differences for arginine, lysine, methionine, threonine, glutamic acid, glycine and tyrosine, and varietal difference for phenylalanene were revealed as significant. The limiting amino acids from the leading varieties were isoleucine, methionine, threonine, alanine and tyrosine, comparing with FAO recommanding levels of amino acids. Among the protein fractions prolamins was the highest in total amino acids, but essential amino acids was the highest in globulins

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Preparation and Characterization of Silicone Hydrogel Lens Containing Poly(ethylene glycol) (PEG를 포함한 실리콘 수화젤 렌즈의 제조 및 특성)

  • Jang, Ha-Na;Chung, Youn-Bok;Kim, Sung-Soo
    • Polymer(Korea)
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    • v.33 no.2
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    • pp.169-174
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    • 2009
  • Silicone hydrogels incorporated with poly(ethylene glycol)(PEG) were prepared and characterized to evaluate the effects of PEG on contact lenses. The silicone hydrogels were copolymerized with methacryloxypropyl tris(trimethylsiloxy) silane (TRIS), methyl methacrylate (MMA), N,N-dimethyl acrylamide (DMA) and PEG-containing monomers such as poly(ethylene glycol) methyl ether methacrylate (PEG- MEM). The silicone hydrogels were characterized using Fourier transform infrared spectroscopy (FT-IR), electron spectroscopy of chemical analysis (ESCA), and scanning electron microscopy (SEM). Water absorbance, water contact angle and light transmittance of the silicone hydrogels were evaluated. The experiments of protein adsorption were also carried out to evaluate the protein adsorption in tears. The peak intensity of C-O bond was increased by the incorporation of PEG-containing monomers and thus PEG incorporation into silicone hydrogels could be confirmed. Phase separation was not shown by the SEM observation of the cross-section of silicone hydrogels. Water absorbancy was increased, while water contact angle and light transmittance were decreased with increasing incorporation of the PEG-containing monomers. The absorption of proteins in tears, albumin, lysozyme and $\gamma$-globulin, on the surface of silicone hydrogels was decreased with increasing incorporation of the PEG-containing monomers.

Isolation of Novel Non-Toxic Bacillus thuringiensis from Soil Samples in Korea (한국 토양으로부터 새로운 무독성 Bacillus thuringiensis 균주의 분리)

  • 노종열;박현우;김호산;진병래;강석권
    • Korean journal of applied entomology
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    • v.34 no.4
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    • pp.373-377
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    • 1995
  • Four Bacillus thuringiensis isolates obtained from soil samples in Korea produce parasporal inclusions non-toxic to 10 insect species of three orders, Lepidopera, Diptera and Coleoptera. These four isolates are named NTB-1, NTB-2, NTB-3 and NTB-4, respectively. The morphology of parasporal inclusions of four isolates observed by phase contrast- and scanning electron microscope was all ovoid. Characterization of four non-toxic B. thuringiensis isolates was determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and restriction endonuclease analysis. The results showed that parasporal inclusion proteins and total plasmid DNA profiles of four isolates are different from other known non-toxic B. thuringiensis strains', suggesting that four isolates are novel.

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Characterization of Plasma Proteins from Bloods of Slaughtered Cow and Pig and Utilization of the Proteins as Adhesives (도축혈액 혈장 단백질의 특성 및 접착제로의 응용)

  • Park, Eun-Hee;Lee, Hwa-Hyoung;Song, Kyung-Bin
    • Applied Biological Chemistry
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    • v.39 no.2
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    • pp.123-126
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    • 1996
  • Simple and rapid method of purification of plasma proteins from bloods of slaughtered animals was developed and the proteins were applied to plywood products as a blood 히ue to utilize waste materials. Plasma protein was obtained by adding 2% trichloroacetic acid (TCA) or 0.6 N HCI as optimal concentration to the supernatant, after centrifugation of bloods. Molecular properties of beef and pig plasma proteins were examined on SDS-PAGE. Application of blood glue to plywood was quite satisfactory compared to the synthetic amino resin by tensile-shear test for the strength of adhesive bonding.

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Characterization of the recombinant cellulase B from Thermotoga maritima (Thermotoga maritima 유래 내열성 cellulase B 융합단백질의 특성 규명)

  • Chung Ho Kim
    • Journal of Applied Biological Chemistry
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    • v.65 no.4
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    • pp.383-386
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    • 2022
  • A gene encoding thermostable cellulase B (TmCelB) was isolated from Thermotoga maritima. The open reading frame (ORF) of TmCelB gene was 825bp long which predicted to encode 274 amino acid residues with a molecular weight of 31,732 Da. The 17 amino acid residues from N-terminal of the TmCelB was known as signal peptides. To analyze the enzymatic activity and biochemical properties, the ORF of TmCelB gene excluding a putative signal sequence encoding 17 amino acids were introduced into the E. coli expression vector, pRSET-B, and overexpressed in E. coli BL21. The optimum temperature of recombinant TmCelB was around 95 ℃, and the optimum pH of recombinant TmCelB was around pH 4.5. The recombinant TmCelB was stable at temperature below 100 ℃.