• Journal of Embryo Transfer
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• v.27 no.1
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• pp.9-14
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• 2012

This study was conducted to analyze the transgenic efficiency and sex ratio in ${\alpha}$-1,3-galactosyltransferase (GalT) knock-out (KO) transgenic pigs according to generation. GalT KO piglets were produced by artificial insemination or natural mating. The transgenic confirmation of GalT KO was evaluated by PCR amplification using specific primers. After electrophoresis, three types of bands were detected such as 2.3 kb single band (Wild), 2.3 and 3.6kb double bands (GalT KO -/+; heterozygote), and 3.6kb single band (GalT KO -/-; homozygote). Transgenic efficiency in F1 generation was 64.5% (23/35) of GalT KO (-/+). In F2 generation, GalT KO transgenic efficiency was 36.4% (21/57, Wild), 47.5% (28/57, GalT KO -/+), and 16.1% (8/57, GalT KO -/-), respectively. Interestingly, no homozygote piglets were born in 6 deliveries among total 11 deliveries, although they were pregnant between male (M) and female (F) $F_1$ heterozygote. In the 5 litters including at least one GalT KO -/- piglet, the transgenic efficiency was 13.3% (2/24, Wild), 51.3% (14/24, GalT KO -/+), and 35.3% (8/24, GalT KO -/-), respectively. The sex ratio of M and F was 40:60 in $F_1$ and 49:51 in $F_2$ generation, respectively. Based on these results, GalT KO transgenic pigs have had a reproductive ability with a normal range of transgenic efficiency and sex ratio.

• KSBB Journal
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• v.9 no.1
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• pp.8-15
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• 1994

The research has been carried out to optimize a recombinant S. cerevisine fermentation process for the production of an anticoagulant hirudin. The structural gene coding for hirudin was combined with the GAL10 promoter for controlled expression, the MFal signal sequence for hirudin secretion, and the GAL7 terminator for transcriptional termination. Growth medium composition and environmental conditions were optimized for maximizing cell growth and final hirudin concentration. The optimized conditions included yeast extract 40g/$\ell$, casamino acid 5g/$\ell$, g1ucose 20g/$\ell$, galactose 30g/$\ell$, DO 50% and temperature $30^{\circ}C$. These conditions yielded the specific cell growth rate of $0.13hr^{-1}$, the final cell density of 30g cell/$\ell$ and the final hirudin concentration of 64mg/$\ell$ in the batch fermentation with a 2.5$\ell$ jar fermentor.

• Proceedings of the Korean Society of Life Science Conference
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• 2002.12a
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• pp.13-19
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• 2002

To elucidate the regulatory mechanism for expression of sialyl-glycoconjugates and their biological functions, ninetheen sialyltransferase cDNAs including eleven by our group or co-works have been cloned and characterized so far. The cloned sialyltransferases are classified into four families according to the carbohydrate linkages they synthesize: ${\alpha}2,3-sialyltransferase$ (ST3Gal I-VI), ${\alpha}$ 2,6-sialyltransferase (ST6Gal I), GalNAc ${\alpha}$ 2,6-sialyltransferase (ST6GalNAc I-VI), and ${\alpha}2,8-sialyltransferase$ (ST8Sia I-VI). Each of the sialyltransferase genes is differentially expressed in a tissue-, cell type-, and stage-specific manner. These enzymes differ in their substrate specificity and various biochemical parameters. However, enzymatic analysis conducted in vitro with recombinant enzyme revealed that one linkage can be synthesized by multiple enzymes. We present here an overview of structure and function of sialyltransferases performed by our group and co-works. Genomic structures and transcriptional regulation of two kinds of human sialyltransferase gene are also presented.

• Food Science of Animal Resources
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• v.42 no.3
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• pp.426-440
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• 2022

The bioactive functions of oligosaccharides from human milk have been reported by many studies. Many of oligosaccharides isolated from colostrum and/or milk of dairy animals have been reported to have similar chemical structures with those in human colostrum and/or milk. It has been proved by several studies that the oligosaccharides with similar chemical structure shared common bioactivities. Among domesticated dairy animals, bovine/cattle, caprine/goat, and ovine/sheep are the most commonly used species to isolate oligosaccharides from their colostrum and/or milk. Several studies on the oligosaccharides from goat colostrum and milk have revealed similar properties to that of human milk and possess the highest content of sialyl oligosaccharides (SOS) as compared to other ruminants. Indonesia ranks first in Association of Southeast Asian Nations (ASEAN) for goat milk production. Therefore, goat milk is the second most consumed milk in the country. The most reared dairy goat breed in Indonesia is Etawah Grade. However, oligosaccharides from Indonesia dairy animals including goat, have not been characterized. This is the first study to characterize oligosaccharides from Indonesia dairy animals. The present study was aimed to isolate and characterize oligosaccharides, specifically SOS from the colostrum of Etawah Grade goats by using proton/¹H-nuclear magnetic resonance. The SOS successfully characterized in this study were: Neu5Ac(α2-3)Gal(β1-4)Glc (3'-N-acetylneuraminyllactose), Neu5Ac(α2-6)Gal(β1-4)Glc (6'-N-acetylneuraminyllactose), Neu5Gc(α2-3)Gal(β1-4)Glc (3'-N-glycolylneuraminyllactose), Neu5Gc(α2-6)Gal(β1-4)Glc (6'-N-glycolylneuraminyllactose), Neu5Ac(α2-6)Gal(β1-4) GlcNAc (6'-N-acetylneuraminyllactosamine) and Neu5Gc(α2-6)Gal(β1-4)GlcNAc (6'-N-glycolylneuraminyllactosamine). This finding shows that Etawah Grade, as a local dairy goat breed in Indonesia, is having significant potential to be natural source of oligosaccharides that can be utilized in the future food and pharmaceutical industries.

• Natural Product Sciences
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• v.13 no.3
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• pp.195-198
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• 2007

Hepatoprotective action of alcoholic extract of Bacopa monniera (BME) was evaluated on Dgalactosamine (D-GalN) induced rat liver toxicity. Bacopa monniera extract reduced the elevated serum enzyme activities of ALT, AST, ALP, LDH, ${\gamma}-GT$ and the formation of hepatic malondialdehyde induced by D-GalN. The alcholic extract of Bacopa monniera also significantly restored the decreased levels of glutathione and the decreased activities of glutathione peroxidase, glutathione reductase, superoxide dismutase, catalase and glucose-6-phosphatase. Therefore these results suggest that Bacopa monniera has hepatoprotective effect against D-GalN induced hepatotoxicity.

• Proceedings of the International Union of Geodesy And Geophysics Korea Journal of Geophysical Research Conference
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• 2003.05a
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• pp.19-19
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• 2003

본 연구에서는 국립해양조사원의 '해양2000호'를 통해 1996년과 1997년에 측정한 동해 지역의 중력자료에 대한 자료 처리를 하였다. 효과적인 자료처리를 위해 선상중력자료 처리에 필요한 각종 보정 절차와 문제점 등을 알아보았으며, 선상자료와 해면고도계자료의 비교 및 이를 통한 선상자료의 검증을 실시하였다. 선상중력자료는 측정과 처리 과정에 있어 여러 사항을 고려하여야 한다. 즉, 육상중력기점을 이용한 절대중력으로의 환산 문제, 선박 항해 위치의 부정확성에 기인하는 문제 및 중력계의 기계적 특성과 중력 측정이 이루어질 때의 해상 조건에 의한 영향 등으로 선상중력자료에 나타나는 여러 오차를 최소화하여야 한다. 선상중력자료로부터 각종 지구중력장 연구에 필요한 중력이상을 계산하기 위해 선상중력 측정시 기인되는 각종 요인의 오차를 고려한 효과적인 보정이 이루어져야 한다. 즉, 선상중력계의 기계변이 보정, 탐사선에 대한 위치 자료의 획득 및 필터링, 그리고 탐사선의 이동으로 인한 Eotvos 효과의 정확한 계산 및 보정이 필요하고, 선상중력계의 기계적 특성에 의해 나타나는 시간지연에 대한 보정도 필요하다. 또한 이러한 보정을 통해 계산한 중력 이상에서 각 교점의 오차를 보정하는 교정오차 보정도 실시하여야 한다. 특히, 탐사선의 이동으로 인한 지구자전 각속도의 상대적인 증감의 효과로 나타나는 Eotvos 효과의 영향은 선상중력자료의 정확도에 가장 큰 영향을 미친다. 이의 정확한 계산 및 보정을 위해서는 정확한 위치정보가 필요하며 본 연구에서는 이를 위해 GPS 항해정보에 대한 Kalman 필터를 실시하였고, Eotvos 효과에 대해 Savitzky-Golay 필터를 적용하여 최적의 Eotvos 보정을 시도하였다. 본 연구에서 계산된 동해지역의 중력이상에 대한 대력적인 범위는 경도 129° - 133°이고 위도 35° - 38.3° 부근이다. 이 지역에 대한 고도이상은 최소 -42.46 mGal에서 최고 161.13 mGal사이에 분포하며, 고도이상의 평균은 14.450 mGal이다. 또한 Bouguer 이상은 최소 -l5.09 mGal에서 최고 218.61 mGal이고 이의 평균은 82.681 mGal이다. 그리고 동해지역의 선상중력 측정지역에서 선상자료에 의한 중력이상과 Altimeter 자료에 의한 고도이상의 전반적인 윤곽은 비슷하면서도 일부 작은 이상의 차이가 나타났으며, 지형자료와 비교하여 보면 Altimeter에 의한 결과보다 선상측정에 의한 결과가 더욱 잘 일치하고 있어 본 연구에서 계산한 선상자료의 타당성을 알 수 있다. 고도이상의 차이는 최소 -25.94 mGal에서 최대 85.33 mGal의 차이를 보이며 차이의 평균은 3.517 mGal, RMS는 6.774 meal이다. 이는 비교적 큰 차이로 선상측정자료의 중요성과 필요성을 단적으로 나타내고 있다.

• Journal of Embryo Transfer
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• v.28 no.3
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• pp.281-287
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• 2013

A major barrier to progress in pig to primate organ transplantation or cell therapy is the presence of terminal ${\alpha}$-1,3-galactosyl epitopes on the surface of pig cells. Therefore, the purpose of this experiment was to establish and cha- racterize mesenchymal stromal/stem cells (MSCs) derived from ${\alpha}$-1,3-galactosyltransferase (GalT) knock out (GalT KO) pig to confirm their potential for cell therapy. Bone marrow (BM)-MSCs from GalT KO pig of 1 month old were isolated by Ficoll-Paque PLUS gradient and cultured with A-DMEM + 10% FBS on plastic dishes in 5% $CO_2$ incubator at 38.5. GalT KO BM-MSCs were analyzed for the expression of CD markers ($CD45^-$, $29^+$, $90^+$ and $105^+$) and in vitro differentiation ability (adiopogenesis and osteogenesis). Further, cell proliferation capacity and cell aging of GalT KO BM-MSCs were compared to Wild BM-MSCs by BrdU incorporation assay (Roche, Germany) using ELISA at intervals of two days for 7 days. Finally, the cell size was also evaluated in GalT KO and Wild BM-MSCs. Statistical analysis was performed by T-test (P<0.05). GalT KO BM-MSCs showed fibroblast-like cell morphology on plastic culture dish at passage 1 and exhibited $CD45^-$, $29^+$, $90^+$ and $105^+$ expression profile. Follow in ginduction in StemPro adipogenesis and osteogenesis media for 3 weeks, GalT KO BM-MSCs were differentiated into adipocytes, as demonstrated by Oilred Ostaining of lipid vacuoles and osteocytes, as confirmed by Alizarinred Sstaining of mineral dispositions, respectively. BrdU incorporation assay showed a significant decrease in cell proliferation capacity of GalT KO BM-MSCs compared to Wild BM-MSCs from 3 day, when they were seeded at $1{\times}10^3$ cells/well in 96-well plate. Passage 3 GalT KO and Wild BM-MSCs at 80% confluence in culture dish were allowed to form single cells to calculate cell size. The results showed that GalT KO BM-MSCs($15.0{\pm}0.4{\mu}m$) had a little larger cell size than Wild BM-MSCs ($13.5{\pm}0.3{\mu}m$). From the above findings, it is summarized that GalT KO BM-MSCs possessed similar biological properties with Wild BM-MSCs, but exhibited a weak cell proliferation ability and resistance to cell aging. Therefore, GalT KO BM-MSCs might form a good source for cell therapy after due consideration to low proliferation potency in vitro.

• Bulletin of the Korean Chemical Society
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• v.13 no.1
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• pp.4-5
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• 1992

• Journal of Acupuncture Research
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• v.17 no.2
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• pp.95-117
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• 2000

The purpose of this morphological studies was to investigate the relation between the meridian, acupoints and viscera using neuroanatomical tracers. The common locations of the spinal ganglia, sympathetic chain ganglia, spinal cord and brain projecting to the large intestine meridian were observed following injection of transganglionic tracer, WGA-HRP and transsynaptic neurotropic virus, pseudorabies virus(PRV), Bartha strain(Ba) and PRV-Ba-Gal (Galactosidase)) into the the large intestine(cecum, colon and rectum), ST37 and LI4. After survival times of 96 hours following injection into the thirty rats with WGA-HRP, PRV-Ba and PRV-Ba-Gal. They were perfused, and their spinal ganglia, sympathetic chain ganglia, spinal cord and brain were frozen sectioned($30{\mu}m$). These sections were stained by HRP and X-gal histochemical and PRV immunohistochemical staining method, and observed with a light microscope. The results were as follows : 1. WGA-HRP labeled neurons innervating the large intestine were observed bilaterally within the T13-L4 sympathetic chain ganglia, and T9-11 spinal ganglia. WGA-HRP labeled neurons innervating ST37 were observed within the L3-5 sympathetic chain ganglia, and L2-4 spinal ganglia. WGA-HRP labeled neurons innervating LI4 were observed in the middle cervical ganglion and stellate ganglion, and C5-8 spinal ganglia. 2. In spinal cord, PRV-Ba labeled neurons projecting to the large intestine, ST37 and LI4 were found in thoracic, lumbar and sacral spinal segments. Densely labeled areas of each spinal cord segment were founded in lamina N, V, VII(intermediolateral nucleus), Ⅸ, X and dorsal nucleus. 3. In medulla oblongata, PRV-Ba and PRV-Ba-Gal labeled neurons projecting to the large intestine, ST37 and LI4 were commonly found in the A1 noradrenalin cells/C1 adrenalin cells/caudoventrolateral reticular nucleus, dorsal motor nucleus of vagus nerve, nucleus tractus solitarius, raphe obscurus nucleus, raphe pallidus nucleus, raphe magnus nucleus and gigantocellular nucleus. 4. In pons, PRV-Ba and PRV-Ba-Gal labeled neurons were commonly found in locus coeruleus, Kolliker-Fuse nucieus and A5 cell group. 5. In midbrain, PRV-Ba and PRV-Ba-Gal labeled neurons were commonly found in central gray matter. 6. In diencephalon, PRV-Ba and PRV-Ba-Gal labeled neurons were commonly found in paraventricular hypothalamic nucleus. These results suggest that PRV-Ba and PRV-Ba-Gal labeled common areas projecting to the large intestine may be correlated to that of the large intestine meridian, ST37 and LI4. Especially, These morphological results provide that interrelationship of meridian-acupoints -viscera may be related to the central autonomic pathways.

• Journal of Sericultural and Entomological Science
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• v.40 no.1
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• pp.52-59
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• 1998

For the construction of plasmid and bmNPV sarrying the FRT recognition site for the FLP recombinases, we synthesized the wild type FRT dligonucleotides. The target FRT sequences consist of three 13bp repeated DNA sequences; two repeats in a direct orientation and one inverted relative to the other two. In addition, there is an 8bp spacer region between the repeats which determune the orientation of the FRT recombination site. In order to place the FRT site both in target BmNPV genome and the transfer vector, we constructed a plasmid, FRT site both in the target BmNPv genome and the transfer vector, we constructed a plasmid, pFRT$\beta$-gal, carrying the FRT sites within the cloning sites of pSV vector and a recombinant BmNPV, vFRTPH, carrying the FRT sites at a downstream of polyhedrin promotor, respectively. In order to test the functionality of the FLP/FRT site-specific recombination system, vFRTPH, pFRT$\beta$-gal and pHsFLP DNA were co-transfected into BmN-4 cells. The resulting recombinant virus was designated a vFRT$\beta$2-gal. From construction analysis of the vFRT$\beta$2-gal with PCR technique it was concluded that the entire pFRT$\beta$-gal plasmid with $\beta$-galactosidase gene and origines of replication flanked by two functional hybrid FRT sequences. The efficiency of recombination was 8.7%, which was higher than that(2.2%) of recombination between a conventional transfer vector and the wild type BmNPV.