• Korean Journal of Microbiology
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• v.43 no.4
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• pp.243-249
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• 2007

The centromere is a highly differentiated structure of the chromosome that fulfills a multitude of essential mitotic and meiotic functions. Alphoid DNA (${\alpha}$-satellite) is the most abundant family of repeated DNA found at the centromere of all human chromosomes, and chromosomes of primates in general. The most important parts in the development of Human Artificial Chromosomes (HACs), are the isolation and maintenance of stability of centromeric region. For isolation of this region, we could use the targeting hook with alphoid DNA repeat and cloned by Transformation-Associated Recombination (TAR) cloning technique in yeast Saccharomyces cerevisiae. The method includes rolling-circle amplification (RCA) of repeats in vitro to 5 kb-length and elongation of the RCA products by homologous recombination in yeast. Four types of $35\;kb{\sim}50\;kb$ of centromeric DNA repeat arrays (2, 4, 5, 6 mer) are used to examine the stability of repeats in homologous recombination mutant strains (rad51, rad52, and rad54). Following the transformation into wild type, rad51 and rad54 mutant strains, there were frequent changes in inserted size. A rad52 mutant strain showed extremely low transformation frequency, but increased stability of centromeric DNA repeat arrays at least 3 times higher than other strains. Based on these results, the incidence of large mutations could be reduced using a rad52 mutant strain in maintenance of centromeric DNA repeat arrays. This genetic method may use more general application in the maintenance of tandem repeats in construction of HAC.

• Biomolecules & Therapeutics
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• v.6 no.1
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• pp.89-94
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• 1998

CJ50001 is a recombinant human granulocyte colony-stimulating facto, (rHuG-CSF) that stimulates the formation of neutrophils from bone marrow stem cells. It was produced in E. colt and purified through refolding and several processes. We produced CS970125(300) using purified C150001 and additives in order to test the stability of CJ50001. When CS970125(300) was stored at 50'S for more than 1 week, high molecular weight proteins were formed and those proteins were detected by non-reducing SDS-PAGE, gel filtration HPLC, and Western blot. Those proteins showed single band at the same position of CJ50001 in reducing SDS-PAGE. These data indicated that those high molecular weight proteins were the multimers of C150001. In biological assays, iu viro and in viro, the multimers did not have biological activity and inhibitory action to that of CJ 50001. The mutimers did not induce toxicity in mice and rats in acute toxicity test. These results suggest that if Cs970125(300) containing CJ50001 is stored at 5$0^{\circ}C$, CJ50001 will be the multimers that do not have biological activity and inhibitory effect to CJ50001 and do not induce acute toxicity.

• Journal of Plant Biotechnology
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• v.36 no.4
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• pp.309-319
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• 2009

Transgenic plant cell cultures for the production of biopharmaceuticals including monoclonal antibodies, recombinant proteins have been regarded as an alternative platform in addition to traditional microbial fermentation and mammalian cell cultures. Plant-made pharmaceuticals (PMPs) have several advantages such as safety, cost-effectiveness, scalability and possibility of complex post-translational modifications. Increasing demand for the quantity and diversity of pharmaceutical proteins may accelerate the industrialization of PMP technology. Up to date, there is no plant-made recombinant protein approved by USFDA (Food and Drug Administration) for human therapeutic uses due to the technological bottlenecks of low expression level and slight differences in glycosylation. Regarding expression levels, it is possible to improve the productivity by using stronger promoter and optimizing culture processes. In terms of glycosylation, humanization has been attempted in many ways to reduce immune responses and to enhance the efficacy as well as stability. In this review article, all these respects of transgenic plant cell cultures were summarized. In addition, we also discuss the general characteristics of plant cell suspension cultures related with bioreactor design and operation to achieve high productivity in large scale which could be a key to successful commercialization of PMPs.

• Molecules and Cells
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• v.25 no.4
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• pp.494-503
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• 2008

Many therapeutic glycoproteins have been successfully generated in plants. Plants have advantages regarding practical and economic concerns, and safety of protein production over other existing systems. However, plants are not ideal expression systems for the production of biopharmaceutical proteins, due to the fact that they are incapable of the authentic human N-glycosylation process. The majority of therapeutic proteins are glycoproteins which harbor N-glycans, which are often essential for their stability, folding, and biological activity. Thus, several glyco-engineering strategies have emerged for the tailor-making of N-glycosylation in plants, including glycoprotein subcellular targeting, the inhibition of plant specific glycosyltranferases, or the addition of human specific glycosyltransferases. This article focuses on plant N-glycosylation structure, glycosylation variation in plant cell, plant expression system of glycoproteins, and impact of glycosylation on immunological function. Furthermore, plant glyco-engineering techniques currently being developed to overcome the limitations of plant expression systems in the production of therapeutic glycoproteins will be discussed in this review.

• BMB Reports
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• v.44 no.10
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• pp.665-668
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• 2011

In order to clarify the impact of Ca-binding sites (Ca1 and 2) on the conformational stability of neutral proteases (NPs), we have analyzed the thermal, pH and organic solvent stability of a NP variant, V189P/A195E/G203D/A268E (Q-mutant), from Salinovibrio proteolyticus. This mutant has shown to bind calcium more tightly than the wild-type (WT) at Ca1 and to possess Ca2. Q-mutant was resisted against autolysis, thermoinactivation and pH denaturation in a Ca-dependent manner and exhibited better activity in organic solvents compared to the WT enzyme. These results imply that Ca1 and Ca2 are important for the conformational stability of NPs.

• Journal of Biomedical Engineering Research
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• v.30 no.6
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• pp.521-528
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• 2009

This study is about to evaluation of postural stability according to characteristics of electrical stimulation on the ankle muscles. We measured body sway(center of pressure, COP) when various parameters of electrical stimulation was applied to ankle muscles in stable and unstable posture. Subjects consisted of 10 young adults, and electrical stimulation was delivered on right and left of tibialis anterior and Achilles tendon. The body sway was measured during electrical stimulation of three duty cycle and frequencies in stable posture and three amplitudes of sensory threshold in unstable posture. Consequently, the COP Shift is higher during electrical stimulation of 1/30(duty ratio) and 100Hz(frequency) in stable posture. In unstable posture, 100% amplitude of sensory threshold induced postural stability. These findings are important for the rehabilitation system of postural stability and the use of electrical stimulation as somatosensory information.

• Molecules and Cells
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• v.27 no.1
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• pp.1-3
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• 2009

Mitotic spindle mediates the segregation of chromosomes in the cell cycle and the proper function of the spindle is crucial to the high fidelity of chromosome segregation and to the stability of the genome. Nucleation of microtubules (MTs) from centrosomes and chromatin represents two well-characterized pathways essential for the assembly of a dynamic spindle in mitosis. Recently, we identified a third MT nucleation pathway, in which existing MTs in the spindle act as a template to promote the nucleation and polymerization of MTs, thereby efficiently amplifying MTs in the spindle. We will review here our current understanding on the molecular mechanism, the physiological function and the cell-cycle regulation of MT amplification.

• Journal of Microbiology and Biotechnology
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• v.7 no.4
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• pp.258-261
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• 1997

For the efficient production of lysophospholipid the hydrolysis of phospholipid using phospholipase $A_1$ from Serratia sp. MK1 was studied in an aqueous-solvent, a two-phase and an emulsion system. Judged on the basis of productivity and the degree of hydrolysis, the yield of lysophospholipid in a two-phase system was found to be better than that obtained in an emulsion system. Among the 13 organic solvents tested phospholipase $A_1$ showed the most efficient catalytic activity and stability in butyl acetate. When 20% phospholipid was used it was completely hydrolyzed in this two-phase system.

• Journal of Microbiology and Biotechnology
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• v.9 no.2
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• pp.132-139
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• 1999

A cell-entrapment technique using compressed air was applied to Bifidobacterium longum KCTC 3128 for the improvement of bifidobacteria viability. The main cell-entrapment matrix used was alginate, and viability improvement of the B. longum entrapped in alginate lattices was monitored along with the effects of other additional biopolymers. A prerequisite for acquiring consistent results was the uniformity of bead size and cell distribution which was achieved by using compressed air and mixing the cell suspension with sterilized alginate powder, respectively. The viability losses of the B. longum entrapped in alginate beads in the presence of three different substances logarithmically increased in relation to the reaction time, and proportionately decreased with an increased alginate concentration and bead diameter. The strongest improvement in B. longum viability was exhibited with a bead containing 3％ alginate and 0.15％ xanthan gum.

• Macromolecular Research
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• v.17 no.12
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• pp.995-1002
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• 2009

Luminescent polynorbornene (PNB)/quantum dot (CdSe@ZnS; QD) composite nanorods and nanotubes were successfully prepared using anodic aluminum oxide (AAO) membranes of various pore sizes as templates. To protect QDs with high quantum yield from quenching during the phosphoric acid treatment used to remove the AAO templates, chemically stable and optically clear norbornene-maleic anhydride copolymers (P(NB-r-MA)) were employed as a capping agent for QDs. The amine-terminated QDs reacted with maleic anhydride moieties in P(NB-r-MA) to form PNB-grafted QDs. The chemical- and photo-stability of QDs encapsulated with PNB copolymers were investigated by photoluminescence (PL) spectroscopy. By varying the pore size of the AAO templates from 40 to 380 urn, PNB/QD composite nanorods or nanotubes were obtained with a good dispersion of QDs in the PNB matrix.