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Production and diagnostic application of monoclonal antibodies against infectious bursal disease virus (IBDV에 대한 단크론항체 생산 및 진단적 응용)

  • Ryu, Min-Sang;Song, Yoon-Ki;Lee, Seung-Chul;Mo, In-Pil;Kang, Shien-Young
    • Korean Journal of Veterinary Service
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    • v.34 no.1
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    • pp.5-12
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    • 2011
  • Infectious bursal disease (IBD) caused by infectious bursal disease virus (IBDV) is a highly contagious viral disease in chicken. It causes heavy economic loss by immune suppression and high mortality. The IBDV, designated Avibirnavirus in the Family Birnaviridae, has a double-stranded RNA genome formed by two segments, segment A and segment B. Segment A encodes a 108 KDa polypeptide that is self-cleaved to produce pVP2, VP3 and VP4, and later pVP2 is cleaved to VP2. The VP2 contains the antigenic regions responsible for elicitation of neutralizing antibodies and VP3 is a major immunogenic protein of IBDV. In this study, monoclonal antibodies (MAbs) specific for IBDV were produced and characterized. All 15 MAbs were specific for IBDV and did not react with other viruses used in this study. The protein specificity of MAbs was determined by comparing the reactivity patterns of each MAb with IBDV VP2 and VP234 recombinant baculoviruses and Western blot analysis. As a result, 7 MAbs (1F5, 2C8, 2F4, 3C7, 4C3, 6F11, 6G5) and 5 MAbs (2A4, 2G2, 3F5, 3G2, 4F10) were specific for VP2 and VP3, respectively. The protein specificity of 3 MAbs (2B8, 3F7, 3F8) were not determined. Five (2C8, 2F4, 4C3, 6F11, 6G5) of the VP2-specific MAbs had a neutralizing activity against IBDV. Some MAbs reacted with IBDV-infected bursa of Fabricius by indirect fluorescence antibody (IFA) and immunohistochemistry (IHC) assay. The MAbs produced in this study would be used for diagnostic reagents for the detection of IBDV infection.

EFFICIENCY OF PROTEIN UTILIZATION OF FORMALDEHYDE TREATED RAPESEED MEAL BY SHEEP AND ITS INFLUENCE ON CATTLE'S PERFORMANCE

  • Liu, J.X.;Wu, Y.M.;Xu, N.Y.;Wu, Z.W.
    • Asian-Australasian Journal of Animal Sciences
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    • v.6 no.4
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    • pp.601-605
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    • 1993
  • This study was purposed to investigate the efficiency of protein utilization of rapeseed meal (RSM) and formaldehyde-treated RSM (TRSM) by sheep, and their influence on performance of growing heifers. Experiment 1 was conducted according to a double $3{\times}3$ Latin square design involving sex yearling Hu Sheep and three experimental diets. All diets contained 600 g ammoniated rice straw and 200 g concentrate mixture. Three dietary treatments were: (1) 100 g RSM + 40 g soybean meal (URUS), (2) 100 g TRSM + 40 g soybean meal (TRUS) and (3) 100 g TRSM + 40 g treated soybean meal (TRTS). Apparent nitrogen digestibility was significantly higher for diet TRUS than that for URUS or TRTS (p<0.05), but with no significant difference between URUS and TRTS (p>0.05). Proportion of nitrogen retention (NR) to the digestible nitrogen intake for diet TRUS and TRTS was 25.57 (p<0.05) and 23.44% (p<0.05) higher than that for URUS respectively. As a result, proportion of NR to nitrogen intake for diet TRUS and TRTS was 34.74 (p<0.05) and 23.78% (p<0.05) higher than that for URUS respectively. Experiment 2 was conducted with 59 Holstein heifers. They were 12-20 months of age at the start of the trial. The experiment was a $2{\times}2$ factorial trial in which the heifers were given the ammoniated rice straw ad libitum and 1.5 kg hay, and supplemented with either RSM or TRSM at a daily allowance of 1.2 or 1.8 kg per day. The live weight gains for heifers receiving 1.2 and 1.8 kg/d of RSM or TRSM were 0.491 and 0.556 or 0.564 and 0.665 kg/d, respectively. The results suggest that formaldehyde treatment can effectively improve the efficiency of protein utilization of rapessed meal and cattle's performance.

Effect of Soy Protein Isolate Coating on Meat Quality of Pork Fresh Cut during Refrigerated Storage

  • Shon, Jin-Han;Kim, Jin-Ho;Eo, Ji-Hyun;Choi, Yong-Hwa
    • Journal of Applied Biological Chemistry
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    • v.55 no.1
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    • pp.27-34
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    • 2012
  • Soy protein isolate (SPI)-based edible coating, with and without carboxymethyl cellulose (CMC), were used to reduce oxidative degradation of cut pork stored at $4^{\circ}C$ for 5 days. The SPI coating reduced (p<0.05) thiobarbituric acid-reactive substances (TBARS) and peroxide value (PV), compared with controls. The inhibition of TBARS and PV for SPI-coated porks with and without CMC, compared with the control was 19.1 and 23.9, and 25.7 and 37.7%, respectively. The SPI coating prevented loss of $L^*$ and $a^*$ values of porks compared to the control. The ability of the SPI coating to provide a moisture barrier for the porks was reduced (p<0.05). The SPI-coated porks with and without CMC reduced moisture loss by 37.3 and 44.6%, respectively, over the control. However, SPI coating of porks did not inhibit the growth of either total plate counts or L. monocytogenes. The result revealed that SPI can effectively be used as a natural antioxidative coating to extend quality and shelf life of pork.

Single-nucleotide polymorphisms in prion protein gene of the Korean subspecies of Chinese water deer

  • Jeong, Hyun-Jeong;Lee, Joong-Bok;Park, Seung-Yong;Song, Chang-Seon;Kim, Bo-Sook;Rho, Jung-Rae;Yoo, Mi-Hyun;Jeong, Byung-Hoon;Kim, Yong-Sun;Choi, In-Soo
    • Korean Journal of Veterinary Research
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    • v.49 no.1
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    • pp.59-62
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    • 2009
  • Susceptibility to chronic wasting disease (CWD) in cervid species has been associated with polymorphisms in the prion protein gene (PRNP). The single nucleotide polymorphisms (SNPs) were found in the PRNP of the Korean subspecies of Chinese water deer via analyses of the DNA sequences obtained from 34 individual deer. Two SNPs were detected at codons 77 and 100. One SNP at codon 77 encoding Glycine was determined to be a silent mutation but the other SNP detected at codon 100 induced an amino acid change, from Asparagine to Serine. The prion protein (PrP) amino acid sequence of the water deer showed 98.8-99.2% homology with those of American elk, white-tailed deer and mule deer. The PrP of the water deer contained amino acid residues closely related with CWD-susceptibility. This study is the first to describe genetic variations in the PRNP of the Korean subspecies of Chinese water deer.

Characterization of the Ryanodine Receptor and SERCA in Fetal, Neonatal, and Adult Rat Hearts

  • Ramesh, Venkat;Kresch, Mitchell J.;Park, Woo-Jin;Kim, Do-Han
    • BMB Reports
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    • v.34 no.6
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    • pp.573-577
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    • 2001
  • The mammalian heart is known to undergo significant mechanical changes during fetal and neonatal development. The objective of this study was to define the ontogeny of the ryanodine receptor/$Ca^{2+}$ release channel and SERCA that play the major roles in excitation-contraction coupling. Whole ventricular homogenates of fetal (F) (19 and 22 days in gestation), postnatal (N) (1 and 7 days postnatal), and adult (A) (5 weeks postnatal) Sprague-Dawley rat hearts were used to study [$^3H$]ryanodine binding and oxalate-supported $^{45}Ca^{2+}$ uptake. For the ryanodine receptor, the major findings were: (1) The ryanodine receptor density, as determined by maximal [$^3H$]ryanodine binding ($B_{max}$), increased 3 fold between the F22 and A periods ($0.26{\pm}0.1$ vs. $0.73{\pm}0.07$ pmoles/mg protein, p<0.01), whereas there was no significant change during the F22 and N1 development phases ($0.26{\pm}0.1$ vs. $0.34{\pm}0.01$). (2) Affinity of the ryanodine receptor to ryanodine did not significantly change, as suggested by the lack of change in the $K_d$ during the development and maturation. For SERCA, changes started early with an increased rate of $Ca^{2+}$ uptake in the fetal periods (F19: $8.1{\pm}1.1$ vs. F22: $19.3{\pm}2.2$ nmoles/g protein/min; p<0.05) and peaked by 7 days (N7) of the postnatal age ($34.9{\pm}2.1$). Thus, we conclude that the quantitative changes occur in the ryanodine receptor during myocardial development. Also, the maturation of the $Ca^{2+}$ uptake appears to start earlier than that of the $Ca^{2+}$ release.

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Functional Expression of Proteomics-guided AfsR2-dependent Genes in Avermectin-producing Streptomyces avermitilis (Avermectin을 생산하는 Streptomyces avermitilis에서의 Proteomics-guided AfsR2-dependent 유전자의 발현)

  • Kim Myung-Gun;Park Hyun-Joo;Im Jong-Hyuk;Kim Eung-Soo
    • Microbiology and Biotechnology Letters
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    • v.34 no.3
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    • pp.211-215
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    • 2006
  • AfsR2 is a global regulatory protein involved in the stimulation of secondary metabolite biosynthesis in various Streptomyces species including avermectin-producing S. avermitilis. Among several AfsR2-dependent genes identified from the comparative proteomics, the polyribonucleotide nucleotidyltransferase (PNP) and the glyceraldehyde-3-phosphate dehydrogenase (GPD) genes were previously proposed to regulate the actinorhodin production in S. lividans upon afsR2 over-expression positively and negatively, respectively. To show the biological significance of the PNP and GPD genes in the S. avermitilis strains, these two genes were functionally expressed in both the wild-type and the avermectin-overproducing mutant strains. The PNP gene expression stimulated secondary metabolite production in the wild-type S. avermitilis ATCC31267, but not in the avermectin-overproducing S. avermitilis ATCC31780. Interestingly, the GDP gene expression stimulated secondary metabolite production by 4-fold in the wild-type S. avermitilis ATCC31267 and by 2.5-fold in the avermectin-overproducing S. avermitilis ATCC31780, respectively. These results suggest that the biological significance of the afsR2-dependent PNP and GPD gene expressions on antibiotic biosynthetic regulation could be significantly different depending on Streptomyces species.

Effect of the addition of protein and lipid on the quality characteristics of Yakju (단백질과 지방첨가가 약주의 품질특성에 미치는 영향)

  • Kang, Ji-Eun;Kim, Jae-Woon;Choi, Han-Seok;Kim, Chan-Woo;Yeo, Soo-Hwan;Jeong, Seok-Tae
    • Food Science and Preservation
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    • v.22 no.3
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    • pp.361-368
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    • 2015
  • In this study, the fermentation characteristics of Yakju were investigated by addition of protein and lipid. These are classified according to raw material (rice, glucose) and inducing substance (rice protein, rice lipid). Alcoholic fermentation occurred at $25^{\circ}C$, after 14 days. The results of this study were as follows: Alcohol content of Yakju with rice protein was higher than those of other samples. The pH and glucose of rice Yakju were detemied to be 4.86~5.13 and 4.17~4.86, respectively. Titratable acid and the total amino acid content of the Yakju with rice protein were the highest among other samples. The optical density contents of the rice Yakju and glucose Yakju were 0.52~0.653 and 0.27~0.61, respectively. The concentration of organic acids in rice Yakju (433.98~519.31 mg%) was higher than that of glucose Yakju (303.76~387.50 mg%). The major organic acid components of the Yakju were succinic, citric, acetic and lactic acids. The nitrogen compound concentrations of rice Yakju (4377.38~10208.06 ppm) was higher than that of glucose Yakju (671.20~9368.93 ppm). The protein odor correlation coefficient was 0.98 (p<0.001) showing a very high correlation coefficient, while lipid odor coefficient showed a negative correlation with -0.038 (p<0.458).

Development of ELISA System for Screening of Specific Binding Inhibitors for Src Homology (SH)2 Domain and Phosphotyrosine Interactions

  • Lee, Sang-Seop;Lee, Kyung-Im;Yoo, Ji-Yun;Jeong, Moon-Jin;Park, Young-Mee;Kwon, Byoung-Mog;Bae, Yun-Soo;Han, Mi-Young
    • BMB Reports
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    • v.34 no.6
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    • pp.537-543
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    • 2001
  • In the present study, an in vitro ELISA system to assess the interaction between Src homology (SH)2 domains and phosphotyrosine that contain peptides was established using purified GST-conjugated SH2 proteins and synthetic biotinylated phosphotyrosine that contain oligopeptides. The SH2 domains bound the relevant phosphopeptides that were immobilized in the streptavidin-coated microtiter plate in a highly specific and dose-dependent manner. The epidermal growth factor receptor (EGFR)-, T antigen (T Ag)-, and platelet-derived growth factor receptor (PDGFR)-derived phosphopeptides interacted with the growth factor receptor binding protein (Grb)2/SH2, Lck/SH2, and phosphatidyl inositol 3-kinase (PI3K) p85/SH2, respectively. No cross-reactions were observed. Competitive inhibition experiments showed that a short phosphopeptide of only four amino acids was long enough to determine the binding specificity. Optimal concentrations of the GST-SH2 fusion protein and phosphopeptide in this new ELISA system for screening the binding blockers were chosen at 2nM and 500nM, respectively. When two candidate compounds were tested in our ELISA system, they specifically inhibited the Lck/SH2 and/or p85/SH2 binding to the relevant phosphopeptides. Our results indicate that this ELISA system could be used as an easy screening method for the discovery of specific binding blockers of protein-protein interactions via SH2 domains.

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Effects of Dietary Protein Levels and Sources on Calcium and Phosphorus Metabolism in Young Korean Women (한국성인 여성의 단백질 섭취수준과 동.식물성 급원이 칼슘 및 인대사에 미치는 영향)

  • 구재옥
    • Journal of Nutrition and Health
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    • v.24 no.2
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    • pp.124-131
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    • 1991
  • This study was to examine the effects of dietary protein levels and the sources of protein(animal and plant) on calcium and phosphorus metabo lism in 10 healthy Korean female adults. The 26­d day study consisted of a 6-day adaptation period and lO-day moderate protein(109 N, 550mg Ca) a and IO-day high protein( 14g N, 570mg Ca) pe­r riod. During the experimental period, the subjects w were divided into two groups, either consuming a animal protein diet(75 % animal protein) or plant protein diet(75 % plant protein). Calcium(300 mg) was supplemented to two subjects of each d dict group for the last 4 days. Feces, urine and diet were analyzed nitrogen. calcium and phos­p phorus. The apparent absorption of calcium was significantly increased as the protein intake was inc­r reased from 60g to 90g in animal protein diet. A Average calcium absorption rate was 30% and 46 % from high and moderate levels of animal p protein diet Urinary calcium excretion was not affected by t the amount of the protein intake in both animal and plant protein diet. There was significantly higher urinary calcium excretion(I34mg) in high I level of animal protein diet than that( 83mg) in h high level of plant protein diet. Calcium balance was improved as the protein intake increased and c calcium was supplemented. Phosphorus absorp­t tion was more efficient in the high animal diet(77.81 %) than in the high plant diet(55-65%). The overall results indicate that an increase of protein and calcium supplement in moderate pro­ttein intake can improve calcium balance due to the increase of calcium absorption.

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Effect of water temperature on protein requirement of Heteropneustes fossilis (Bloch) fry as determined by nutrient deposition, hemato-biochemical parameters and stress resistance response

  • Fatma, Shabihul;Ahmed, Imtiaz
    • Fisheries and Aquatic Sciences
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    • v.23 no.1
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    • pp.1.1-1.14
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    • 2020
  • Background: Dietary protein requirements are dependent on a variety of factors and water temperature is one of the most important abiotic factors affecting protein requirement of fish. This study was, therefore, conducted to investigate effects of water temperature on dietary protein requirement of fry Heteropneustes fossilis which has high demand in most of the Asian markets. Methods: Quadruplicate groups of 30 fish per treatment (2.97 ± 0.65 cm; 5.11 ± 0.34 g) were fed seven isoenergetic diets (17.9 kJ g-1 gross energy; 14.99 kJ g-1 digestible energy) containing dietary protein levels ranging from 28 to 52% at two water temperatures (18 and 26 ℃). Experimental diets were fed to apparent satiation as semi-moist cakes thrice daily at 17:00, 12:00, and 17:30 h for 12 weeks. For precise information, various growth parameters, protein deposition, hematological parameters, metabolic enzymes, and stress response were analyzed, and effects of water temperature on dietary protein requirement was recommended on the basis of response from above parameters. Results: Groups held at 26 ℃ attained best growth, feed conversion, and protein deposition at 44% dietary protein indicating that temperature affected dietary protein requirement for optimum growth of H. fossilis fry and protein requirement seems to be satisfied with 44% dietary protein. Interestingly, interactive effects of both dietary protein levels and temperature were not found (P > 0.05). Fish reared at 18 ℃ had comparatively higher values for aspartate and alanine transferases than those reared at 26 ℃ water temperature which exhibited normal physiological value for these enzymes indicating that body metabolism was normal at this temperature. Hematological parameters also followed same pattern. Furthermore, fish reared at 26 ℃ water temperature exhibited more resistant to thermal stress (P < 0.05). The 95% maximum plateau of protein deposition data using second-degree polynomial regression analyses exhibited dietary protein requirement of fry H. fossilis between 40.8 and 41.8% of diet at 26 ℃ water temperature. The recommended range of dietary protein level and protein/digestible energy ratio for fry H. fossilis is 40.8-41.8% and 27.21-27.88 mg protein kJ-1 digestible energy, respectively. Conclusions: Information developed is of high significance for optimizing growth potential by making better utilization of nutrient at 26 ℃ and, to develop effective management strategies for mass culture of this highly preferred fish species.