• Title/Summary/Keyword: 박유전

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Molecular Cloning and Gene Expression of Sinorhizobium meliloti Mannitol Dehydrogenase in Escherichia coli, and Its Enzymatic Characterization (Sinorhizobium meliloti 유래 Mannitol Dehydrogenase 유전자의 클로닝 및 대장균 내 발현과 효소특성 규명)

  • Jang, Myoung-Uoon;Park, Jung-Mi;Kim, Min-Jeong;Lee, So-Won;Kang, Jung-Hyun;Kim, Tae-Jip
    • Microbiology and Biotechnology Letters
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    • v.41 no.2
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    • pp.153-159
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    • 2013
  • A mannitol dehydrogenase (MDH; EC 1.1.1.67) gene was cloned from the Sinorhizobium meliloti 1021 (KCTC 2353) genome and expressed in Escherichia coli. It was seen to have an open reading frame consisting of 1,485 bp encoding 494 amino acids (about 54 kDa), which shares approximately 35-55% of amino acid sequence identity with some known long-chain dehydrogenase/ reductase family enzymes. The recombinant S. meliloti MDH (SmMDH) showed the highest activity at $40^{\circ}C$, and pH 7.0 (D-fructose reduction) and pH 9.0 (D-mannitol oxidation), respectively. SmMDH could catalyze the oxidative/reductive reactions between D-mannitol and D-fructose in the presence of $NAD^+/NADH$ as a coenzyme, but not with NADP+/NADPH. These results indicate that SmMDH is a typical $NAD^+/NADH$-dependent mannitol dehydrogenase.

섬유소 분해효소 유전자 도입 형질전환 돼지 생산

  • 이연근;박진기;민관식;성환후;임기순;양병철;김진회;류재웅;장원경
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.58-58
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    • 2001
  • 본 연구는 섬유소 분해효소 유전자 Cel D(Cellulase Digestion)가 도입된 형질전환 돼지 생산을 통하여 섬유소 함유 사료의 이용 효율을 증대시키고, 나아가 장기이식동물 및 고가의 의료용 단백질 생산가축을 개발하기 위한 원천기술 확보에 있다. 섬유소분해 유전자의 크로닝 및 조직 특이적 발현벡터를 개발하기 위하여, 우선 소의 위중 제4위 내의 미생물로부터 전체 유전자를 분리하였고, 이렇게 작성된 DNA library에서 섬유소분해 관련 유전자인 약 2.0 kb의 Cel D유전자를 크로닝하였으며, 췌장 특이적 발현 프로모터(rat elastase I: 약 200bp)를 크로닝한 후, 미세주입용 형질전환 재조합 벡터를 구축하기 위하여 rElastase I 프로모터 하류에 섬유소 분해 유전자(Cel D)를 연결하여 약 3.0 kb 크기의 재조합 벡터를 준비하였으며, 재조합 유전자를 1세포기 수정란 전핵내에 미세주입 하기 위해 Sal I과 BglII를 이용 유전자 단편을 만들었다. 구축된 유전자를 미세주입하기 위한 수정란을 회수하기위해 총68두의 돼지를 4-5두씩 분리사육하면서 발정동기화 및 과배란 유기를 위해 PG600, Altrenogest, FSH, hCG를 투여하였으며 hCG투여후 약54시간에 외과적 방법에 의해 총 1,359개의 수정란을 회수하였고, 이중 미세주입가능한 1세포기 수정란은 1,296개로 두당 평균 15.9개 였다. 1,296개의 1세포기 수정란 중에서 재조합 유전자(rE I-CelD)가 미세주입된 660개의 수정란을 32두의 수란돈에 외과적 방법에 의해 이식하였으며, 이식되어진 모돈 13두가 분만하여 40.6%의 임신율을 나타내었다. 이렇게 분만된 13두에서 총 65두(암:33두, 수:32두)의 자돈이 생산되었으며, 형질전환 여부를 판명하기 위해 자돈의 꼬리조직으로 부터 genomic DNA를 추출하고 PCR 검정을 실시하였다. PCR 검정 결과, 섬유소 분해 유전자가 도입된 자돈은 5두 이었으며, 그 결과를 Table에 나타내었다.(Table Omitted) Table 1 에서와 같이 섬유소 분해효소유전자가 형질전환된 자돈은 65마리 중 5마리로 7.69%의 형질전환율을 나타내었으며, 5마리의 자돈중 2두(암:1두, 수:1두)는 분만 후 즉시 폐사되었으며 2두(암:1두, 수:1두)는 86일령 그리고 14일령에 폐사하여 현재 1두(암)가 생존하여 섬유소 분해 사양 실험 중에 있다.

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A Study for Mechanism and Oxidation Reaction of Substituted Benzyl Alcohols by 2,2'-Bipyridinium Chlorochromate (2,2'-Bipyridinium Chlorochromate에 의한 치환 벤질 알코올류의 산화반응과 메카니즘에 관한 연구)

  • Kim, Young-Sik;Park, Young-Cho
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.13 no.7
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    • pp.3252-3260
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    • 2012
  • 2,2'-Bipyridinium chlorochromate[$C_{10}H_8N_2HCrO_3Cl$] was synthesized by the reaction of 2,2'- bipyridine with chromium(VI) trioxide in 6M HCl. The structure was characterized by IR and ICP analysis. The oxidation of benzyl alcohol using 2,2'-bipyridinium chlorochromate in various solvents showed that the reactivity increased with the increase in the order of the dielectric constant(${\varepsilon}$), in the order: cyclohexene< chloroform$p-CH_3$, H, m-Br, $m-NO_2$) in N,N'-dimethylformamide. Electron-donating substituents accelerated the reaction, whereas electron acceptor groups retarded the reaction. The Hammett reaction constant(${\rho}$) was -0.64(303K). The oxidation reactivity of alcohols can be a useful factor to study about physical properties such as thermal stability, when the polysilsesquioxane solution is ready for an applying coating agent. The observed experimental data was used to rationalized the hydride ion transfer in the rate-determining step.

Two Cases of Gitelman's Syndrome Diagnosed by Renal Clearance Study (신장청소검사를 이용하여 진단한 Gitelman 증후군 2례)

  • Kim, Tae Hwa;Kim, Seung Jun;Seo, Yu Kyung;Shim, Jung-Yeon;Jung, Hye Lim;Park, Moon Soo;Kum, Dong Hyuk
    • Clinical and Experimental Pediatrics
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    • v.45 no.3
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    • pp.413-417
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    • 2002
  • Gitelman's syndrome is an autosomal recessive disorder characterized by hypokalemic metabolic alkalosis, hypomagnesemia, and hypocalciuria that has recently been reported to be linked to thiazide- sensitive Na-Cl cotransporter gene mutation. In this study, we performed renal clearance studies to differentiate Gitelman's from Bartter's syndrome and to confirm the diagnosis in two patients clinically diagnosed with Gitelman's syndrome. Each patient was hydrated by 20 mL/kg body weight of oral water within 30 minutes, which was followed by intravenous half saline. When urinary flow reached 10 mL/min, samples of urine and serum were obtained to calculate the osmolar clearance, free water clearance, chloride clearance, and distal fractional chloride reabsorption. Subsequently, furosemide or hydrochlorothiazide was administered. Samples were collected and the same parameters were calculated. In our patients, chloride clearance was increased more than 10 times after furosemide administration(2.1 : 25.7 and 2.2 : 27.4 mL/min/100 mL GFR), but not increased after hydrochlorothiazide treatment(2.1 : 1.6 and 2.2 : 2.6 mL/min/100 mL GFR). And the distal fractional chloride reabsorption was significantly decreased by furosemide injection (73% : 15% and 75% : 4.6%), whereas hydrochlorothiazide had no effect on it(73% : 63% and 75% : 78%). These findings indicate that our patients have a defect in thiazide-sensitive Na-Cl cotransporter in the distal tubule, which is compatible with the pathophysiology of Gitelman's syndrome.

Evaluation of Pepper Germplasm and Cultivar Introduction (고추 보존(保存) 유전자원(遺傳資源)의 특성평가(特性評價)와 품종도입(品種導入))

  • Lee, Woo Sung;Kim, Byung Soo;Kim, Sang Gee;Park, Kyu Hwan;Kwon, Young Seok
    • Current Research on Agriculture and Life Sciences
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    • v.8
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    • pp.35-44
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    • 1990
  • Sixty two lines of pepper including Korean local cultivars and introductions were planted out to the experiment farm of Kyungpook National University for seed increase, and 16 plant and fruit characters including days to flower were recorded. In days to flower, KC85(Chilgok Dongmyung) and KC116(PI308791), KC154(Cherry Sweet), and KC185(Choseongochu) were the earliest with 83 days, and KC139(Serrano chili) and KC103(PI297488) were the latest with 126 days. In fruit detachment force, KC85(Chilgok dongmyung) was the tenderest with 0.45kg and KC162(PI22444) was the toughest with 3.34kg. In powdery mildew incidence, KC10(Pimiento), KC23(PI224423), KC51(PI257044), KC56(PI257053), KC126(PI358812), KC119(PI322719), KC143(Emerald Giant), KC148(VR-2), KC163(PI224445), KC163(Papari Mild) were the least diseased. Anthracnose was least on KC39(Serrano Chili). Collection and introduction of pepper germplasm was continued. Fourteen Korean local cultivars and 25 foreign cultivars including 2 wild species were additionally collected or introduced.

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The Use of Aureobasidin A Resistant Gene as the Dominant Selectable Marker for the Selection of Industrial Yeast Hybrid (산업용 효모 Hybrid의 선별을 위한 우성선별표지로서의 Aureobasidin A 내성유전자의 이용)

  • Jeon, Han-Taek;Park, Uhn-Mee;Kim, Keun
    • Microbiology and Biotechnology Letters
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    • v.39 no.2
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    • pp.111-118
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    • 2011
  • For the strain improvement of the industrial polyploid yeast strain through hybridization and protoplast fusion, a dominant selection marker other than a recessive marker such as the auxotrophic marker was required for the selection of the resulting hybrids. In the present investigation, the aureobasidin A resistant gene was tested in relation to whether it can be used as the dominant selectable marker for the isolation of hybrids of the yeast Saccharomyces. The plasmid pAUR112, carrying the gene responsible for resistance to aureobasidin A, was introduced into the haploid yeast strain K114/YIp. From the rare-mating between polyploid C6 and haploid K114/YIp carrying pAUR112, many hybrids were obtained from the agar medium containing 0.5 ${\mu}g$/ml of aureobasidin A. The hybrids exhibited characteristics derived from both of the parental strains; and the cell sizes of the hybrids were larger than those of the parental strains. These results showed that the aureobasidin A resistant gene could be successfully used as the dominant selectable marker for the isolation of yeast hybrids resulting from rare-mating.

Effect of Enamel Matrix Drivatives application on the expression of PDLs17, PDLs22 of cultured human periodontal ligament cells in vitro (Enamel Matrix Derivatives가 사람 치주인대 세포의 특이유전자인 PDLs17, PDLs22의 발현에 끼치는 효과)

  • Han, Geun-A;Jang, Hyun-seon;Kok, Jung-Ki;Park, Ju-Chol;Kim, Heoung-Jung;Kim, Jung-Gwan;Kim, Byung-Ock
    • Journal of Periodontal and Implant Science
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    • v.34 no.2
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    • pp.333-344
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    • 2004
  • The enamel matrix derivative (EMD) has been recently used in the periodontal regenerative techniques. The present study was established to investigate the influence of EMD on human periodontal ligament cells using expression of mRNA of periodontal ligament specific gene (PDLs)17, PDLs22, type I collagen when EMD applied to periodontal ligament cells. Periodontal ligament cells were obtained from a healthy periodontium and cultured in Dulbecco's modified Eagle's medium (DMEM) plus 10% fetal bovine serum and ${\beta}-glycerophosphate$ with ascorbic acid. Test groups were two; One adds EMD in culture media and another added EMD and Dexamethasone (DEX) in culture media. Positive control group added DEX in culture media, and negative control group adds niether of EMD nor DEX. $Emdogain^{(R)}$ (Biora, Sweden, 30 mg/ml) was diluted by 75 ${\mu}g/ml$ concentration to culture media. For reverse transcription-polymerase chain reaction (RT-PCR), total RNA isolated on days 0, 7, 14 and 21. mRNA of PDLs17 was expressed on days 14 and 21 in EMD or DEX group, and expressed on days 7, 14 and 21 in EMD plus DEX group, the other side, expressed on days 21 in negative control group. mRNA of PDLs22 expressed on days 7, 14 and 21 in EMD group, and expressed on days 14 and 21 in DEX group, and expressed on days 7, 14 and 21 in EMD plus DEX group. Negative control group expressed on days 14 and 21. Type I collagen was expressed on all days and all groups. These results indicate that EMD promotes differentiation of periodontal ligament cells, and this is considered to offer basis that can apply EMD to periodontal tissue regeneration technique.

Profiling of Metabolites and Proteins from Eschscholtzia californica induced by Yeast Extract (Yeast Extract로 처리된 Eschscholtzia californica의 Metabolite와 Protein의 변화)

  • Cho Hwa-Young;Park Jeong-Jin;Yoon Sung-Yong;Part Jong Moon
    • KSBB Journal
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    • v.20 no.4
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    • pp.285-290
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    • 2005
  • Benzophenanthridine alkaloids - sanguinarine, chelirubine, macarpine, and chelerythrine are produced from Eschscholtzia californica (Californica Poppy, used as a sedative by Native Americans) and most of them are derived from dihydrosanguinarine. The properties of sanguinarine are the basis of its antimicrobial activity and its use in chemosurgery and skin cancer excision. For overproduction of sanguinarine from E. californica, yeast extract was used as elicitor and the elicited cell's metabolites were checked. Sanguinarine production was increased intracelluarly about 8 times in the cell and 5 times extracelluarly. We have peformed proteomic analysis of proteins sequentially extracted from E. califormica suspended cells which were cultured with elicitor, an increase of spot intensity was seen at 24 hours following elicitation. These proteins were separated by two-dimensional electrophoresis (2-DE). We found several spots that were expected to be related to benzophenanthridine alkaloids production by comparing the production profiles of metabolites such as sanguinarine. These results demonstrate the use of metabolite analysis as a tool for detecting target proteins related to metabolites production pathway.

Estimation of the effective population size using single-nucleotide polymorphism (SNP) information in Korean Holstein dairy cattle (단일염기다형성 정보를 이용한 국내 홀스타인 젖소의 유효집단 크기 추정)

  • Cho, Kwang-Hyun;Do, Kyoung-Tag;Park, Kyung-Do
    • Journal of the Korean Data and Information Science Society
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    • v.28 no.3
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    • pp.597-604
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    • 2017
  • In this study, we investigated the genetic characteristics and the effective population size of domestic dairy cattle using 42,201 SNPs for 923 heads of Holstein cattle. The estimate for the average linkage disequilibrium ($r^2$) among the adjacent SNPs by chromosome was 0.22, and it was highest (0.26) in chromosome 14 and lowest (0.17) in chromosome 27. When the physical distance among SNPs was less than 25Kb, the estimate for the average $r^2$ was $0.31{\pm}0.33$ and it was markedly decreased as the physical distance increased. When the physical distance among SNPs was larger than 25Mb, the estimate for the average $r^2$ was 0.04, and it decreased by 0.27 (87.1%) compared with case of physical distance of less than 25Kb. There was a trend that the effective population size in Holstein dairy cattle decreased over generations and the estimate for the effective population size in the first 5 generations (1~5th generation) was 110 heads.

DISPLACEMENT OF MAXILLARY LATERAL INCISOR CAUSED BY IDIOPATHIC GINGIVAL FIBROMATOSIS (특발성 치은 섬유종증에 의한 상악 측절치의 변위)

  • Jung, Ji-Sook;Park, Ho-Won;Lee, Ju-Hyun;Seo, Hyun-Woo;Lee, Suk-Keun
    • Journal of the korean academy of Pediatric Dentistry
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    • v.38 no.3
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    • pp.296-302
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    • 2011
  • Idiopathic gingival fibromatosisrarely occurs, but frequently recurred after surgical removal. It usually occurs in generalized symmetrical pattern but sometimes in localized unilateral pattern. The localized pattern usually affects the maxillary molar and tuberosity area. This disease usually causes tooth migration, malocclusion, and problems in eating, speech, and esthetics. A boy showed dense gingival fibromatosis localized at primary maxillary right lateral incisor area at the age of 5 years, and his maxillary right lateral incisor become severely displaced at the age of 9 years. He had no medical and hereditary factors relevant to the gingival fibromatosis. However, the dense fibrous tissue was dominant in his labial gingiva of maxillary right incisors. In order to realign the displaced incisors by orthodontic treatment, the dense fibrous tissue covered the defect space between the central incisor and the displaced lateral incisor was surgically removed. The removed specimen was examined by simple immunohistochemical(IHC) array method. IHC array showed increased expression of CTGF, HSP-70, MMP-1, PCNA, CMG2, and TNF-${\alpha}$ in keratinocytes, fibroblasts, endothelial cells, and macrophages of gingival fibromatosis tissue. Therefore, it was suggested that the gingival fibromatosis be caused by the concomitant overexpression of CTGF, HSP-70, MMP-1, PCNA, CMG2, and TNF-${\alpha}$, and resulted in the fibroepithelial proliferation and the inflammatory reaction of gingival tissue.