• Journal of Microbiology and Biotechnology
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• v.15 no.2
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• pp.254-258
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• 2005

We have conducted in vitro reconstitution study of ferritin from its subunits FerH and FerL. For the reconstitution, FerH was produced from an expression vector construct in Escherichia coli and was purified from a heat treated cell extract by using one-step column chromatography. FerL was expressed as inclusion bodies. The denatured form of FerL was obtained by a simple washing step of the inclusion bodies with 3 M urea. The reconstitution experiment was conducted with various molar ratios of urea-denatured FerH and FerL to make the ferritin nanoparticle with a controlled composition of FerH and FerL. SDS-PAGE analysis of the reconstituted ferritins revealed that the reconstitution required the presence of more than 40 molar$\%$ of FerH in the reconstitution mixture. The assembly of the subunits into the ferritin nanoparticle was confmned by the presence of spherical particles with diameter of 10 nm by the atomic force microscopic image. Further analysis of the particles by using a transmission electron microscope revealed that the reconstituted particles exhibited different percentages of population with dense iron core. The reconstituted ferritin nanoparticles made with molar ratios of [FerH]/[FerL]=l00/0 and 60/40 showed that 80 to $90\%$ of the particles were apoferritin, devoid of iron core. On the contrary, all the particles formed with [FerH]/[FerL]=85/ 15 were found to contain the iron core. This suggests that although FerH can uptake iron, a minor portion of FerL, not exceeding $40\%$ at most, is required to deposit iron inside the particle.

• KSBB Journal
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• v.26 no.2
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• pp.119-125
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• 2011

In the synthesis of nanoparticles, much attention has been paid to regulating the particle size. There has been a possible evident that using the central cavity (core) of the protein ferritin has a greatly significant influence on it because the core can generate the nanometer-sized mineral particles of variable metal ions. In this report, recombinant human L-ferritins produced from Saccharomyces cerevisiae were purified and their molecular properties were characterized. The cDNA for human ferritin L chain was also expressed in another host such as Escherichia coli, and the properties of recombinant L-ferritins were compared. From isoelectric focusing experiment, the L-ferritin from the recombinant yeast showed no indication of N-glycosylation. Some post-translational modifications other than N-glycosylation were speculated in the L-ferritins from yeast. A difference was made in the L-ferritins in their iron uptake rates and the initial rate of the L-ferritin from yeast was slightly increased. The reconstitution yield and size distribution of the core minerals were analyzed in the L-ferritins by transmission electron microscopy. The L-ferritin from yeast with higher reconstitution yield (54.5%) showed slightly larger sizes (mean 6.92 nm) with narrower size distribution than the L-ferritin from E. coli. It is, in conclusion, speculated that L-ferritin from yeast is relatively superior to the other, in view of the size of nanoparticle and its relative homogeneity.

• Journal of Embryo Transfer
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• v.15 no.3
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• pp.247-253
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• 2000

This study was undertaken to optimize enucleation and reconstitution methods for the production of cloned mice by somatic cell nuclear transfer Outbred ICR mouse oocytes at the metapahse- II stage were retrieved from female mice superovulated by PMSG and hCG. In Experiment 1, oocytes were enucleated in medium supplemented with cytochalasin B (CCB) of 3 levels (0, 7.5 or 15 $\mu\textrm{g}$/mL), and higher rate of encleation was obtained at 7.5 and 15 $\mu\textrm{g}$/mL than at $\mu\textrm{g}$/mL. In Experiment 2, oocytes enucleated in 7.5 $\mu\textrm{g}$/mL CCB-containing medium were reconstituted with different types of somatic cell by following methods; 1) cumulus cells by direct cell injection, 2) cumulus cells by electric fusion (1.25 kV/cm, 2 pulses for each 70 $mutextrm{s}$) or 3) STO cells by the electrofusion. Electrofusion of STO cells with enucleated oocytes yielded the greatest (P<0.05) rate of reconstitution without lysis (76%) than any other combinations. Although significant decrease in the rate of somatic cell introduction was found, the electrofusion of cumulus cells yielded better rate of reconstitution than direct injection (0 vs. 18%). In Experiment 3, the duration of electric stimulation for the fusion was changed to either 50 $mutextrm{s}$ or 90 $mutextrm{s}$, but no significant improvement of reconstitution efficacy was obtained. In conclusion, this study showed that ICR mouse oocytes could be used for the production of reconstituted oocytes and a fusion method of 1.25 KV/cm with 2 pulses using 570 cell was the optimal.

• Asian-Australasian Journal of Animal Sciences
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• v.18 no.4
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• pp.538-544
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• 2005

The present experiment was conducted to assess the effect of reconstitution (R) on utilization of red sorghum (S) in diets of broiler chickens. Day-old broiler chicks (n=360) were randomly divided into 36 groups of 10 chicks each, and 9 dietary treatments were allotted to 4 groups (replicates) in a completely randomized design. Out of the 9 treatments, one was corn-soy based control (D1). The rest of the treatments were diets consisting of four levels (25, 50, 75 and 100% part of corn) of raw red sorghum (S25-S100) or four levels of reconstituted red sorghum (RS25-RS100). The tannin content reduced from 2.3% to 1.6% after reconstitution of red sorghum. Body weight gain reduced significantly (p<0.01) in diets containing unprocessed red sorghum beyond 33% in diet or reconstituted red sorghum at any level. However, during finishing growth phase the birds receiving either processed or unprocessed sorghum (barring S75) had statistically similar gains in body weight. During over all growth phase (0-6 wk), live weight gains in all the dietary treatments did not differ statistically. Feed intake during 0-6 wk was significantly higher (p<0.05) in diets containing sole red sorghum than corn-soy based control diets Feed conversion ratio during 0-3 wk period in control and unprocessed red-sorghum diets were similar but statistically poorer (p<0.01) FCR emanated from reconstituted groups, while during 3-6 wk of age FCR was poorer (p<0.05) in diets containing 75% red sorghum, either processed or unprocessed. FCR, in overall growth phases, in control diet was statistically similar to the groups fed diets containing up to 33% unprocessed or 16% reconstituted group. The carcass traits and yield of organs did not differ (p>0.05) due to the various levels of red-sorghum. It was concluded that though the tannin content was reduced by 30% by the reconstitution process, but this did not give any additional advantage in broiler performance. More over, red-sorghum can be used effectively up to 33% in diet replacing 50% of corn after proper adjustment of proteins, energy and amino acids.

• Proceedings of the KOSOMBE Conference
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• v.1996 no.05
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• pp.103-107
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• 1996

In this paper, We propose an algorithm for reconstitution of chromosome images to extract its morphological feature parameters. It is reconstituted from 460 chromosome images using the 32 direction line algorithm. We extract three morphological feature parameters such as centromeric index, relative length ratio, and relative area ratio. The experiment results show that our method is batter than that of other researchers comparing with the error of feature parameters.

• BMB Reports
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• v.35 no.3
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• pp.255-261
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• 2002

GTP cyclohydrolase I (E.C. 3.5.4.16) is a homodecameric protein that catalyzes the conversion of GTP to 7,8-dihydroneopterin triphosphate (H2NTP), the initial step in the biosynthesis of pteridines. It was proposed that the enzyme complex could be composed of a dimer of two pentamers, or a pentamer of tightly associated dimers; then the active site of the enzyme was located at the interface of three monomers (Nar et al. 1995a, b). Using mutant enzymes that were made by site-directed mutagenesis, we showed that a decamer of GTP cyclohydrolase I should be composed of a pentamer of five dimers, and that the active site is located between dimers, as analyzed by a series of size exclusion chromatography and the reconstitution experiment. We also show that the residues Lys 136, Arg139, and Glu152 are of particular importance for the oligomerization of the enzyme complex from five dimers to a decamer.

• Korean Journal of Heritage: History & Science
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• v.48 no.1
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• pp.148-165
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• 2015

Iron was one of the most influential factors for formation and development of ancient countries. The diffusion of ironware had increased agricultural productivity and brought about military technical revolution. Needless to say, the rise and fall of the countries depended on the possession of stable iron production. Raw materials and fuels are the key factors for mass production of iron and a transportation route is essential to supply the goods. Jungwon area satisfies the three factors. There are many iron manufacture sites such as Jincheon Seokjang-ri Gusan-ri, and Chunju Chilgeum-dong Tangeumdae earthen ramparts in the Jungwon area. In order to study the ancient iron manufacture technique, reconstitution experiment was carried out using restored furnace which was made based on the Jincheon Seokjang-ri B-23 furnace. Some notable results were identified with the experiment as in the followings. Firstly, a roasting process has a connection with the decrease of hardness of the iron ore. Secondly, melting of the blast pipe as well as the formation of product within the furnace had a crucial effect on the cessation of the experiment. Thirdly, reduced iron in various locations within the furnace prove that there was enough reducing environment during the working. Not only melting point but also properties of iron can vary depending on the carbon contents. For the reason, formation of approximate environment in which iron can react to the chalcoal is the most important factor in terms of iron manufacture.

• Korean Journal of Heritage: History & Science
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• v.48 no.4
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• pp.138-153
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• 2015

A Jincheon Seokjangri B23 furnace was reconstructed and iron smelting experiment was performed to investigate an ancient Baekjae iron production process. The work mainly described in this paper is the $1^{st}$ and $2^{nd}$ experiments among the several experiments carried out at Jungwon National Research Institute of Cultural Heritage. Iron ore(magnetite) and oak charcoal were used as a source and a foot bellow was used for air supply. Common results of the experiments are masses of iron, slag and charcoal formed in the furnace. Most iron lumps were formed nearby the tuyere rather than the area of tapping hole. Metallographic and chemical analysis shows that the iron lumps can be used for either forge or cast depending on their carbon content. Low Fe content and glassy texture of the inner slags suggest that the operation environment was quite reducing. Based on the results of the iron smelting experiments, measurements and analysis, various information was obtained regarding physical-chemical and metallurgical processes of the ancient iron smelting process. It is firmly believed that its undisclosed contents can be revealed more in depth with continual reconstitution experiments.