• Title/Summary/Keyword: gels

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Translation Initiation Factor IF1-Dependent Stimulation of 30 S Preinitiation Complex Formation: Rapid Isolation and fMEt-tRNA Binging Activity of IF1

  • CHOIK, SANG-YUN;HYUN-JUNG KIM;JUNG-IK YANG;HYO-IL CHANG
    • Journal of Microbiology and Biotechnology
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    • v.11 no.6
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    • pp.986-993
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    • 2001
  • Translation Initiation in prokaryotes involves the formation of a 30 S preinitiation complex, in which translation initiation factors play a role in the stimulation of fMet-tRNA (fMet) binding. However, the specific function and precise mechanism of initiation factor IF1 are still unclear. One a functionally active factor with a high purity. In the present study a large quantity of active IF was rapidly purified, obtained by the overexpression of the infA gene, and then used for a functional study. The induction of infA did not appreciably affect the growth rate of the protease-deficient strain E. coli AR68 harboring the IF1 overproducing plasmid. The level of IF1 obtained was approximately $1-2\%$ of the total cell protein, which enabled the yield of highly purified IF1 (>$98\%$ pure) to be increased to 0.15 mg of IF1/g of cells. The IF1 was isolated within one day by the centrifugatioin of the ribosomal washed fraction, by ammonium sulfate fractionation, chromatography on batch of phosphocellulose, and FPLC Mono S. The overexpressed IF1 was found to be comparable to the factor isolated from normal cells, as determined by migration in NEPHGE/SDS 2-D gels. For binding of fMet-tRNA(fMet) to the 30 S ribosomal subunitis, relatively high levels of binding were obtained when IF2 was present. The addition of IF1 up to 110 pmol proportionally stimulated the binding to a variable extent. This IF1-dependent stimulation of the 30 S preinitiation complex formation demonstrated that IF1 would appear to be exclusively essential for promoting the initiation phase of protein synthesis.

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Gel Color and Texture of Surimi-like Pork from Muscles at Different Rigor States Post-mortem

  • Kang, Geun-Ho;Yang, Han-Sul;Jeong, Jin-Yeon;Moon, Sang-Hoon;Hur, Sun-Jin;Park, Gu-Boo;Joo, Seon-Tea
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.7
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    • pp.1127-1134
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    • 2007
  • Gels were made from surimi-like pork (SLP) made from muscles obtained at 1, 24 and 72 h post-mortem. The SLP from pre-rigor muscle had higher pH and moisture percentage compared to in- or post-rigor muscles. Also, SLP from pre-rigor muscle showed higher concentration of water-soluble protein that was washed out during the process. Gel from post-rigor muscle exhibited higher a* and b* value, and also resulted in higher Chroma and lower hue values. The dark color of gel from post-rigor muscle was related to higher concentration of sarcoplasmic protein in SLP and denser structure in the gel matrix. SDS-PAGE showed higher intensity of the phosphorylase in the sarcoplasmic protein fraction from pre-rigor muscle. Gel from post-rigor muscle showed higher hardness and sensory firmness, and the greater firmness was related to higher concentration of protein in SLP, and a compact network with smaller pockets in the gel matrix.

Skeletal Ryanodine Receptor 1-Heterozygous PSE (Pale, Soft and Exudative) Meat Contains a Higher Concentration of Myoglobin than Genetically Normal PSE Meat in Pigs

  • Obi, T.;Matsumoto, M.;Miyazaki, K.;Kitsutaka, K.;Tamaki, M.;Takase, K.;Miyamoto, A.;Oka, T.;Kawamoto, Y.;Nakada, T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.9
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    • pp.1244-1249
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    • 2010
  • Comparisons of properties between skeletal ryanodine receptor 1 (sRyR1)-heterozygous-mutated and normal types of meat were carried out in pigs using PSE (pale, soft and exudative) meat found during the butchering process. All samples considered to be PSE meat showed irregular running and disorder of the muscle fibers and a wider inter-fiber space upon light microscopic observation. Electron microscopy revealed disintegration, twisting, and disorder of the myofibril arrangement and elimination of the Z line in PSE meat, compared with normal meat. Meat property tests demonstrated greater decreases in water holding capacity, moisture and sarcoplasmic protein, and higher $L^*$ values for the meat color index in PSE meat than in normal meat, but there were no differences in these factors between genetically normal and sRyR1-heterozygous PSE meat. On the other hand, higher $a^*$ and $b^*$ values were observed in sRyR1-heterozygous than in normal PSE meat, and similar alterations to the a* value were observed in terms of the amount of myoglobin and density of the 17-kDa protein band, corresponding to the molecular mass of myoglobin, on SDS-PAGE gels. These results suggest that sRyR1-heterozygous PSE pork contains much more myoglobin than genetically normal PSE meat.

The New LM-PCR/Shifter Method for the Genotyping of Microorganisms Based on the Use of a Class IIS Restriction Enzyme and Ligation-Mediated PCR

  • Krawczyk, Beata;Leibner-Ciszak, Justyna;Stojowska, Karolina;Kur, Jozef
    • Journal of Microbiology and Biotechnology
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    • v.21 no.12
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    • pp.1336-1344
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    • 2011
  • This study details and examines a novel ligation-mediated polymerase chain reaction (LM-PCR) method. Named the LM-PCR/Shifter, it relies on the use of a Class IIS restriction enzyme giving restriction fragments with different 4-base, 5' overhangs, this being the Shifter, and the ligation of appropriate oligonucleotide adapters. A sequence of 4-base, 5' overhangs of the adapter and a 4-base sequence of the 3' end of the primer(s) determine a subset of the genomic restriction fragments, which are amplified by PCR. The method permits the differentiation of bacterial species strains on the basis of the different DNA band patterns obtained after electrophoresis in polyacrylamide gels stained with ethidium bromide and visualized in UV light. The usefulness of the LM-PCR/Shifter method for genotyping is analyzed by a comparison with the restriction endonuclease analysis of chromosomal DNA by the pulsed-field gel electrophoresis (REA-PFGE) and PCR melting profile (PCR MP) methods for isolates of clinical origin. The clustering of the LM-PCR/Shifter fingerprinting data matched those of the REA-PFGE and PCR MP methods. We found that the LM-PCR/Shifter is rapid, and offers good discriminatory power and excellent reproducibility, making it a method that may be effectively applied in epidemiological studies.

Varietal Classification of Barley by Isozymes and Restriction Fragment Length Polymorphisms (RFLPs) (동위효소 분석과 제한효소 단편 다형화현상을 이용한 보리 품종의 분류)

  • Jin, Byung-Soon;Park, Ro-Dong;Eun, Moo-Young;Lee, Eun-Seop
    • Applied Biological Chemistry
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    • v.36 no.3
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    • pp.139-145
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    • 1993
  • The methods of isoelectric focusing of 4 isozymes in polyacrylamide horizontal slab gels and restriction fragment length polymorphisms (RFLPs) were applied to characterize the biochemical phenotypes of 19 cultivars of barley. Among 19 barley cultivars screened, 7 esterase, 3 phosphoglucose isomerase, 4 peroxidase and 2 alcohol dehydrogenase isozyme phenotypes were distinguished by isoelectric focusing. When purified DNA of each cultivar was digested with restriction enzyme EcoRV and analyzed its RFLPs with barley DNA markers pMSU 51 or pMSU 71, two distinct RFLP patterns were shown. Based on the four isozymes and two RELP polymorphisms, 19 cultivars of barley were classified into 13 biochemical phenotypes. Phylogenetical relationships among 13 biochemical phenotypes classified were determined using Nei's F-statistics and the phylogenetic dendrogram was constructed.

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Effects of Processing Conditions on the Protein Quality of Fried Anchovy Kamaboko Engraulis japonica

  • Ramos, Leny R. Ordonez;Choi, Nam-Do;Ryu, Hong-Soo
    • Fisheries and Aquatic Sciences
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    • v.15 no.4
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    • pp.265-273
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    • 2012
  • The effects of processing and frozen storage conditions on the quality of anchovy Engraulis japonica fried surimi gels were investigated. Protein content decreased after surimi gel processing from 19.6% (raw meat) to 12.1% (kamaboko) due to the added ingredients and change in water content. Lipid content decreased from 2.8% (raw meat) to 1.3% in minced and 0.5% in surimi, but fried kamaboko showed a 6.9 % lipid level. Thiobarbituric acid values and thiobarbituric acid reactive substances levels were highest in kamaboko samples, 89.5 and 1.9 mg/g solid, and increased gradually with storage time to 101.8 and 4.6 mg/g solid, respectively. In vitro protein digestibility increased from 79.2% in raw anchovy to 88.5% in kamaboko samples. Levels of trypsin inhibitor decreased gradually with processing and during storage time from 2.43 in raw anchovy to 0.31 mg/g solid in the kamaboko sample after 60 days of frozen storage. No noticeable changes in total essential amino acid was observed during processing conditions. Computed protein efficiency ratio for kamaboko was highest (2.59) compared with whole anchovy (1.96), minced (1.94) and surimi (2.50). Fresh fried anchovy kamaboko showed similar values of hardness, springiness, gumminess and chewiness to commercial surimi gel, but a higher values were seen for fracturability and adhesiveness, and lower values for cohesiveness and resilience. The frozen and thawed anchovy kamaboko showed higher values for all of these rheological parameters compared with fresh and commercial kamaboko. Anchovy kamaboko showed the lowest lightness (62.9) and redness (0.16) and similar yellowness (11.9) compared with commercial kamaboko. Frozen storage and vacuum packaging were effective maintaining the shelf life of anchovy kamaboko within 30 days, but were not effective after 45 days due to fat oxidation.

Synthesis and Characterization of Biodegradable Elastic Hydrogels Based on Poly(ethylene glycol) and Poly(${\varepsilon}-caprolactone$) Blocks

  • Im, Su-Jin;Choi, You-Mee;Subramanyam, Elango;Huh, Kang-Moo;Park, Ki-Nam
    • Macromolecular Research
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    • v.15 no.4
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    • pp.363-369
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    • 2007
  • Novel biodegradable elastic hydrogels, based on hydrophilic and hydrophobic polymer blocks, were synthesized via the radical crosslinking reaction of diacrylates of poly(ethylene glycol) (PEG) and poly(${\varepsilon}-caprolactone$) (PCL). PEG and PCL diols were diacrylated with acryloyl chloride in the presence of triethylamine, with the reaction confirmed by FT-IR and $^1H-NMR$ measurements. The diacrylate polymers were used as building-blocks for the syntheses of a series of hydro gels, with different block compositions, by simply varying the feed ratios and molecular weights of the block components. The swelling ratio of the hydrogels was controlled by the balance between the hydrophilic and hydrophobic polymer blocks. Usually, the swelling ratio increases with increasing PEG content and decreasing block length within the network structure. The hydrogels exhibited negative thermo-sensitive swelling behavior due to the coexistence of hydrophilic and hydrophobic polymer components in their network structure, and such thermo-responsive swelling/deswelling behavior could be repeated using a temperature cycle, without any significant change in the swelling ratio. In vitro degradation tests showed that degradation occurred over a 3 to 8 month period. Due to their biodegradability, biocompatibility, elasticity and functionality, these hydrogels could be utilized in various biomedical applications, such as tissue engineering and drug delivery systems.

Effect of P(MAA-co-PEGMA) Hydrogel Compositions and Loading pH on the Loading Efficiency of Cosmetic Active Agents (P(MAA-co-PEGMA) 수화젤의 조성과 탑재 pH가 화장품 활성물질의 탑재효율에 미치는 영향)

  • Lee, Eun-Mi;Kim, Kyu-Sik;Kim, Bum-Sang
    • Polymer(Korea)
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    • v.33 no.5
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    • pp.441-445
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    • 2009
  • In order to develop an intelligent delivery system for cosmetic active agents (e.g., arbutin, ascorbic acid, and adenosine), pH-responsive P(MAA-co-PEGMA) hydrogel particles having an average size of approx. $2{\mu}m$ and spherical shape were synthesized via dispersion photopolymerization. There was a drastic change in the swelling ratio of P (MAA-co-PEGMA) hydro gels at a pH 5, which is the $pK_a$ of the hydrogel. To determine the factors that have an effect on the loading of cosmetic active agents into the hydrogel, the loading efficiency of the active agents was investigated at various MAA and EG compositions of the hydrogel and the loading pH conditions. When the MAA contents in the hydrogel decreased, the loading efficiency of the active agents increased. In loading experiments with different pH conditions, the active agents showed higher loading efficiency into the hydrogel at the pH where the hydrogel was at the swollen state than at the pH where the electrostatic repulsion was minimized.

Wound Healing Effect of Curcumin Gel for Transdermal Delivery (경피전달을 위한 커큐민 젤의 창상치유효과)

  • Kim, Jin;Kim, Man Jong;Lee, Ki-Young
    • Polymer(Korea)
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    • v.37 no.3
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    • pp.387-392
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    • 2013
  • The aim of this study was to investigate the effect of trasdermal delivery of the curcumin gel on healing of the rats' dorsum wounds. Carbopol 934 and propylene glycol were used to prepare gels containing 1% curcumin. Curcumin gel was evaluated for various properties such as antioxidant, cell viability, anti-inflammatory, in vivo wound healing. The free radical scavenging activity using 1,1-diphenyl-2-picryl hydrazyl (DPPH) was 50% at 12.5 ppm concentration. Inhibition of nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells by curcumin gel. In the curcumin gel-treated group, the re-epithelialization in wounds was significantly increased compared to the control group throughout the experimental period. These results suggested that curcumin may be helpful for the promotion of wound healing.

Identification of Species and Sex of Korean Roe Deer (Capreolus pygargus tianschanicus) Using SRY and CYTB Genes

  • Han, Sang-Hyun;Cho, In-Cheol;Lee, Sung-Soo;Tandang, Leoncia;Lee, Hang;Oh, Hong-Shik;Kim, Byoung-Soo;Oh, Moon-You
    • Animal cells and systems
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    • v.11 no.2
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    • pp.165-168
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    • 2007
  • The nucleotide sequences of a male-specific marker sex determining region Y (SRY) gene and a mitochondrial cytochrome B (CYTB) gene were characterized and analyzed to establish a molecular method for identification of species and sex of Korean roe deer (Capreolus pygargus tianschanicus). Similarity search result of SRY sequences showed very similar result to those reported in Moose (Alces alces) and Reindeer (Rangifer tarandus), both of which had 95.9% similarity in identity. CYTB genes were very similar to those reported in Siberian roe deer (C. pygargus pygargus) which had 98.6% similarity and not to European roe deer (C. capreolus), suggesting that the DNA samples tested were of Siberian roe deer lineage. Polymerase chain reaction (PCR)-based sex typing successfully discriminated between carcasses of male and female roe deer. Males had SRY band on agarose gels and females did not. The result of this molecular sex typing provided similar information with that obtained by genital organ observation. Therefore, this molecular method using male specific marker SRY and mitochondrial CYTB genes would be very useful for identification of the species and sex of the carcass remains of roe deer.