• Title/Summary/Keyword: Protein determination

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Backbone NMR Assignments of WW2 domain from human AIP4

  • Seo, Min-Duk
    • Journal of the Korean Magnetic Resonance Society
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    • v.24 no.2
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    • pp.38-42
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    • 2020
  • WW domains are small protein modules consisting of three-stranded antiparallel β-sheet, and involved in the protein-protein interaction for various biological systems. We overexpressed and purified WW2 domain from human AIP4/Itch (a member of Nedd4 family) using a pH/temperature dependent cleavage system. The backbone assignments of WW2 domain were completed, and secondary structure was predicted. Furthermore, backbone flexibility of WW2 domain was determined by 1H-15N heteronuclear NOE and amide hydrogen exchange experiments. The structural information would contribute to the structural determination of WW2 domain as well as the interaction study of WW2 domain with various binding partners.

Trend and Technology of Gene and Genome Research (유전자 및 유전체 연구 기술과 동향)

  • 이진성;김기환;서동상;강석우;황재삼
    • Journal of Sericultural and Entomological Science
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    • v.42 no.2
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    • pp.126-141
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    • 2000
  • A major step towards understanding of the genetic basis of an organism is the complete sequence determination of all genes in target genome. The nucleotide sequence encoded in the genome contains the information that specifies the amino acid sequence of every protein and functional RNA molecule. In principle, it will be possible to identify every protein resposible for the structure and function of the body of the target organism. The pattern of expression in different cell types will specify where and when each protein is used. The amino acid sequence of the proteins encoded by each gene will be derived from the conceptional translation of the nucleotide sequence. Comparison of these sequences with those of known proteins, whose sequences are sorted in database, will suggest an approximate function for many proteins. This mini review describes the development of new sequencing methods and the optimization of sequencing strategies for whole genome, various cDNA and genomic analysis.

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Chemical and Physical Predictors of the Nutritive Value of Wheat in Broiler Diets

  • Ball, M.E.E.;Owens, B.;McCracken, K.J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.1
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    • pp.97-107
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    • 2013
  • The aim of this study was to establish relationships between chemical and physical parameters of wheat with performance and digestibilities of feed components in broiler chickens fed on wheat-based diets. Ninety-four wheat samples were selected for inclusion in four bird trials. Birds were housed in individual wire metabolism cages from 7 to 28 d and offered water and feed ad libitum. Dry matter intake (DMI), liveweight gain (LWG) and gain:feed were measured weekly. A balance collection was carried out from 14 to 21 d for determination of apparent metabolizable energy (AME), ME:gain, dry matter retention, oil and neutral detergent fibre (NDF) digestibility. At 28 d the birds were humanely killed, the contents of the jejunum removed for determination of in vivo viscosity and the contents of the ileum removed for determination of ileal dry matter, starch and protein digestibility. When wheat parameters were correlated with bird performance data, it was found that specific weight was not significantly (p>0.05) related to bird performance. Bird DMI, LWG and gain:feed were best correlated (p<0.05) with the rate of starch digestion, although the coefficients of correlation (r) were still low (0.246 to 0.523). A negative relationship (p<0.01) between AME and total (r = -0.432) and soluble (r = -0.304) non starch polysaccharide (NSP) was observed in this study. Thousand grain weight (TG) was positively correlated with DMI (r = 0.299), LWG (r = 0.343) and gain:feed (r = 0.371). When establishing multiple regression relationships, correlation coefficients greater than 0.8 were achieved for DMI, LWG, gain:feed and ileal crude protein digestibility. However, the economics involved in determining the parameters involved in the regressions make the process impractical.

Characterization of a Low Molecular Weight Heat-Shock Protein cDNA Clone from Nicotiana tabacum

  • Park, Soo-Min;Joe, Myung-Kuk;Hong, Choo-Bong
    • Proceedings of the Botanical Society of Korea Conference
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    • 1999.04a
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    • pp.18-18
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    • 1999
  • We characterized a cDNA clone for a low molecular weight heat-shock protein (LMW HSP) from tobacco named TLHS-l. Nucleotide sequence determination of TLHS-1 identified an open reading frame for 159 amino acids. To the upstream of the open reading frame, a sequence of 124 nucleotides was determined. To the 3' downstream of the open reading frame, 212 nucleotides were identified which carried poly(A)-tail. Comparison of the open reading frame and hydropathy plot of TLHS-1 with the previously reported class I LMW HSPs showed high identity which classified TLHS-1 as a class I LMW HSP cDNA clone. We proposed that there are six consensus regions in class I LMW HSPs. RNA blot hybridization for TLHS-1 showed a typical expression pattern of heat-shock-inducible gene from three common tobacco cultivars. The open reading frame of TLHS-1 was overexpressed in Escherichia coli. TLHS-1 protein confers thermal protection of other proteins in vitro and in vivo. Thermal induced aggregation of citrate synthase was reduced by purified TLHS-1 protein, and thermal death rate at $50^{\circ}C$ was reduced in E. coli expressing TLHS-l. From these data, we can expect that TLHS-1 acts as a molecular chaperone.perone.

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Development of Crystallization Distinction Supporting System Using Image Processing

  • Saito, Kanako;Kawabata, Kuniaki;kunimitsu, Satoshi;Asama, Hajime;Mishima, Taketoshi
    • Proceedings of the IEEK Conference
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    • 2002.07c
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    • pp.1788-1791
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    • 2002
  • In the post-genome era. it is one of important research subject to Investigate the roles of the proteins in human body based on decoded genome information during Human Genome Project. In order to clarify them. it is necessary to analyze the structure of the protein crystals and their function. ' Crystallization is the beginning stage of protein structure determination process. There are some methods for structural analysis of the proteins, and general one is X-ray structural analysis method. In order to utilize this method for analyzing the protein crystal's structure, artificial protein crystallization is required. However, since artificial crystallizing work takes much time and manpower. the performance against its cost is still low. Therefore. we started to discuss to develop a supporting system for improving efficiency of the crystallization distinction procedure. In this paper, we examine to realize such supporting system for crystallization distinction using image-processing technique and report about our experimental result with many real protein solution images.

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Determination of the Synthetic Time and the Transport Pattern of Vicilin and Legumin in Ginseng Endosperm Cell Using Double Immunogold Labeling (이중 면역금입자 표지법을 이용한 인삼 배유세포내 Vicilin과 Legumin의 합성시기 및 수송방식)

  • Lee, Chang-Seob;Yu, Seong-Cheol;Kim, Woo-Kap
    • Journal of Ginseng Research
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    • v.19 no.3
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    • pp.267-274
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    • 1995
  • Vicilin and legumin, the storage Proteins of seed, were Purified from ginseng (Panax ginseng C.A. Meyer) endosperm cells. They were immunized in rabbits, and antibodies were raised respectively. Using these two antibodies, double immunogold labeling of vicilin and legumin was carried out to determine the gap of synthetic time and the transport pattern of vicilin and legumin in the ginseng endosperm cells. Vicilin and legumin were synthesized at the same time at early embryo developmental stage. They were secreted from the Golgi bodies and accumulated into the small vacuoles. As the endosperm cells developed, vicilin and legumin localized in the small vacuoles were gradually transported toward the large central vacuole where they were stored. Protein bodies were derived from the vacuoles filled with proteins and distributed in the endosperm cells of mature red seed. Protein bodies were various in size from 1 to 8 ${\mu}{\textrm}{m}$ in which vicilin and legumin were mixed each other. The number of small particles labeled on the vicilin was greater than that of large particles labeled on the legumin in the protein bodies indicating that the amount of vicilin is higher than that of legumin in the protein bodies.

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Determination of Protein Requirements for Maintenance of Elk Doe

  • Kim, Kwan-Woo;Lee, Jinwook;Kim, Dong-Gyo;Lee, Eun-Do;Lee, Sung-Soo;Kim, Do Hyung;Lee, Sang-Hoon
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.40 no.3
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    • pp.177-181
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    • 2020
  • Data on the crude protein requirements of elk doe are nonexistent and the data are essential for their management in Korea. Therefore, this study was conducted to evaluate the crude protein requirement for maintenance of elk doe. Three female elk deer were used in 3 × 3 Latin square design with three diets containing three levels of crude protein (CP) that contained low crude protein (approximately 12%), medium crude protein (15%), and high crude protein (18%). Each three elk doe trials included a 14-day preliminary period and a 5-day collection period. Crude protein intake was 4.83, 6.26, and 9.00 g/d for 12%, 15%, and 18% of CP level, respectively. Crude protein balances were 1.04, 1.41, and 4.14 for 12%, 15%, and 18% of CP level, respectively. The maintenance requirement for CP from the regression equation between CP intake and CP balance were 3.70 g/BW0.75.

Identification of Limiting Amino Acids and Determination of Requirement of Total Sulfur-containing Amino Acids in a Low Protein Diet in Young Chicks. (어린병아리에서 저단백질사료내 제한아미노산의 규명과 함유황아미노산의 요구량 결정)

  • Chee, Kew-Mahn
    • Korean Journal of Poultry Science
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    • v.11 no.1
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    • pp.1-12
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    • 1984
  • Since a 13% dietary protein level is generally accepted as a standard in evaluating net protein utilization values of protein sources in chicks, limiting amino acids a 13% protein basal diet containing 15% isolated soy-protein as the only source of dietary protein, were identified. Of such amino acids as methionine, lysine, threonine and tryptophan added to the basal diet singly or as a combination, methionine appeared as the only limiting amino acid for optimum growth of the chicks. When the requirement of total sulfur-containing acids (TSAA) was estimated as the point at which the dose-response curve intersected a line representing the plateau for maximum performance, the TSAA requirements for maximum growth and feed intake were 4.73% and 3.73% of dietary protein, respectively. The values, expressed in terms of TSAA intake, required for maximum weight gain, feed intake and gain/feed ratio were 167.1, 136.8 and 159.1 mg/bird/day, respectively.

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Rapid Evaluation of Chemical Components of Rice Grain Using Near Infrared Spectroscopy (근적외분광분석법에 의한 미질관련 성분 측정)

  • 황흥구;조래광;손재근;이수관
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.39 no.1
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    • pp.7-14
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    • 1994
  • This study was conducted to establish the rapid evaluation method of chemical components of rice grain on the basis of non-destructive method. A near-infrared reflectance spectroscopic(NIRS) method was utilized, for the determination of amylose, protein, magnesium, and potassium content of rice. A multiple linear regression analysis for the data obtained by standard laboratory methods and NIRS method was carried out to make a calibration. The standard error of prediction for amylose, protein, magneisum and potassium content were 0.88%, 0.28%, 12.62mg and 10.79mg, respectively. It was concluded that the NlRS method can be useful the rapid determination of amylose, protein, magnesium and potassium content instead of the existing laboratory method.

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Evaluation of the Kit's Efficiency of Alpha Fetoprotein (AFP) Test (Alpha feto protein(AFP)검사 키트의 유효성 평가실험)

  • Cho, Hyun-su;Noh, Gyeong-woon;You, Tae-min
    • The Korean Journal of Nuclear Medicine Technology
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    • v.22 no.2
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    • pp.101-104
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    • 2018
  • Purpose Alpha fetoprotein(AFP) is a fetal serum protein that increases in germ cell tumors derived from liver cancer or egg yolk. AFP test has been used for screening of liver cancer, determination of tumor stage, determination of therapeutic effect, and fetal congenital malformations. The purpose of this study was to compare the results of the four kits, identify the advantages and disadvantages of each kit, and select the appropriate kits for our laboratory. Materials and Methods Blood samples were obtained from 89 patients attending the Seoul national university hospital. Experiments were carried out in accordance with manufacturer's instructions of four companies(A, B, C, D). The precision, recovery, linearity, and sensitivity test were performed for each kit. Results In case of the precision within the measurement, the CV value of the C kit was less than 5% at the low, middle, and high concentrations. The A, B and D kit's the CV value was less than 5% at the concentrations except the low concentration. The recovery rates of the A, B, C, and D kits were $100{\pm}15%$, $100{\pm}30%$, $100{\pm}16%$ and $100{\pm}14%$, respectively. All kits showed good linearity. Sensitivity was measured as 0.5 IU/mL for A, 0.4 IU/mL for B, 0.98 IU/mL for C, and 0.3 IU/mL for D. Conclusion The CV values of the four kits were within 10%, and the correlation coefficients were close to 1 for $R^2=0.978$, $R^2=0.992$ and $R^2=0.8957$. As a result, they are clinically available. Therefore, each laboratory should select the appropriate kit for their experiment's environment.