• Title/Summary/Keyword: 형질 세포

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Functional Analysis of Gene ID1103135 Encoding a 3-Phytase Precursor Homologue of Streptomyces coelicolor (Streptomyces coelicolor의 3-Phytase 상동성 유전자 ID1103135의 기능분석)

  • 김미순;강대경;이홍섭;연승우;김태영;홍순광
    • Korean Journal of Microbiology
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    • v.40 no.2
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    • pp.81-86
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    • 2004
  • Among the annotated ORFs of Streptomyces coelicolor, SCO7697 was supposed to encode for phytase (myo-inositol hexakisphosphate phosphohydrolase). The DNA fragment containing SCO7697 was cloned by the PCR from the chromosomal DNA of S.coelicolor A3(2)M. The cloned fragment was introduced into E. coli expres-sion vector, pET28a(+), to yield two recombinant plasmids, pET28-SP and pET28-LP, which were designed to encode different length of proteins. When the pET28-SP and pET28-LP were introduced into E. coli BL21, the transformants successfully overexpressed recombinant proteins, but the molecular weights of the expressed pro-teins were appeared bigger than those of expected in SDS-polyacrylamide gel electrophoresis. The shift of cul-tural temperature from 37 to $30^{\circ}C$ made most of expressed protein be solubilized. The expressed protein, however, did not show any phytase activity. When the DNA fragment with its own promoter placed on the E. coli-Streptomyces vector, pWHM3, and introduced into S. lividans, the phytase activity was not detected either. These results suggest that even though the SCO7697 was annotated as a probable phytase with high probability (E value is $6e^{-89}$), the real product doest not have phytase activity.

Ecological and Morphological Characteristics of Zostera caulescens Miki (Zosteraceae) in Korea (한국산 수거머리말(Zostera caulescens Miki.)의 형태 및 생태적 특성에 대한 연구)

  • Lee, Sang-Yong;Suh, Young-Bae;Kim, Sang-Tae;Choi, Chung-Il
    • Ocean and Polar Research
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    • v.24 no.4
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    • pp.345-357
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    • 2002
  • Ecological characteristics on habitats and morphological features of the seagrass Zostera caulescens Miki in Korea were examined. The biogeographical distribution of Z. caulescens was confirmed in the south coast of the Korea. Zostera caulescens usually inhabits at the inner bay, where is sheltered from wave action and 3.0 to 6.5 m deep. The sediment in habitats is composed of very fine muddy sand or sandy mud. In morphology, Z. caulescens is easily distinguished from other species of the genus by the formation of vegetative canopy on the top of reproductive stems. The number of longitudinal ribs in testa was 24 revealed by scanning electron microscope (SEM) while the number of ribs in Z. marina has been often reported to be 16-20. Purplish anthocyanin spots were displayed on the surface when the testa was removed. We found that the size and shape of leaf epidermal cells in Z. caulescens were very different from those of Z. marina when the leaf cuticles were removed by maceration which could be useful characters for identifying Zostera species. The leaf of Z. caulescens displays two different casts of color when the fresh plant is closely observed. The margins of leaves appear brighter than the center of leaves due to the thickening by the development of paralleled venation in the middle of leaves. The comparison of two populations of Z. caulescens in Korea showed that they were considerably different in their shoot density and biomass. The shoot density and biomass at Gabae population was $367.3 m^{-2}\;and\;725.7g$ dry wt $m^P{-2}$ respectively, while those at Jukrim population were $112.5m^{-2}\;and\;392.0g\;dry\;wt\;m^{-2}$, respectively.

Information Requirements, IS Planning Typology, and IS Planning Effectiveness : An Empirical Study (정보요구, 정보시스템 기획유형, 그리고 정보시스템 기획 효과성에 관한 실증적 연구)

  • 문태수
    • Proceedings of the Korea Association of Information Systems Conference
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    • 1997.10b
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    • pp.111-136
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    • 1997
  • 정보시스템 기획(IS planning)은 경영환경의 변화로부터 야기되는 조직의 정보요구 를 제대로 파악함으로써 새로운 정보시스템의 구축이나 실행과정에 반영하는 중요한 과업중 의 하나이다. 하지만, 아직까지 정보요구를 반영한 정보시스템 기획유형에 관한 연구나 정 보시스템 기획의 효과성에 관한 영향에 대한 실증적인 연구가 없었다. 그리하여, 본 연구에 서는 기업의 경쟁력 강화 목표를 달성하기 위해 필요한 정보시스템의 효율적인 관리와 활용 을 위하여 정보시스템 기획에 관한 상황 이론적 접근을 시도하였다. 본 연구는 과업환경을 중심으로 발생하는 정보요구의 유형에 따라 정보시스템 계획을 어떻게 추진하는 것이 바람 직한 것인지를 파악하고, 정보시스템을 기획 시 정보요구 수준과의 적합성(fitness)에 따라 정보시스템 기획의 효과성은 어떻게 달라지는 지를 파악하고자 한다. 이를 위하여 본 연구 에서 사용된 Construct와 변수들은 기존의 정보시스템 기획관련 문헌을 이용하였다. 본 연 구를 위하여 국내 매출액 1000대 기업을 대상으로 설문지를 우송하였으며, 그 중 187부가 회수되었다. 자료분석의 결과, 정보요구 수준과 정보시스템 기획유형간의 관련성은 긍정적 으로 관련되어 있었으며, 정보요구 수준에 따른 기획의 효과성에는 차이가 없었으나, 정보 시스템 기획유형에 따른 기획의 효과성은 유의한 차이가 있었다. 특히, 정보요구 수준과 정 보시스템 기획유형간의 적합성에 의한 정보시스템 기획의 효과성에 대한 t-test결과는 적합 한 조직(fit organization)보다는 적합하지 않은 조직(unfit organization)의 성과가 더 높은 것으로 나타났다.성 형질전환 계통의 식물체 내에서의 바이러스 증식 및 이동과 관련된 저항성 기작, 여러 가지 PVY 계통에 대한 저항성 유무, 수량, 생육 특성 및 주요 화학 성분 함량 등을 발표하고자 한다.-glucose로 구성된 다당류 이었다. 아미노산은 Asp 및 Glu의 산성 아미노산과 Ala, Leu 등의 함량이 높게 나타났으며, 비알칼리 추출물에서 Ser과 Thr의 함량이 높게 나타났다. 다당류 T-AS는 평균 분자량이 2,000 kD와 12kD에서 주 peak를 나타냈으며, 수용성 분획의 평균 분자량은 12kD이고 비수용성 분획은 36~2,000 kD의 평균 분자량 분포를 갖는 것으로 나타났다. IR과 NMR 분석 결과 890 cm-1에서 흡수 peak를 나타내어 $\beta$-(1,3)0glucan과 $\beta$-(1,6)-glucan의 구조를 갖는 다당류로 확인 되었다. T-AS 분획은 C:H:O:N의 함량비가 38.9:5.7:49.6:1.84%이며, 이 물질의 융점은 163 $^{\circ}C$로 연한 갈색을 나타낸다. 분리된 GLG의 항암활성 기전 규명을 위해, in vivo 항암실험, 항보체 활성능, 항체 생성능, serum protein 분비능, 대식세포의 탐식능과 활성능 및 세포간 물질 분비 등의 상관관계를 조사하였다. 다당류 GLG 분획물들 가운데 항보체의 활성이 높았던 분획은 sarcome 180에 대한 항암 활성이 높게 나타났다. 다당류 T-AS의 보체 활성화 기작은 classical과 alternative complement pathway의 양 경로를 통해 활성화 되었다. T-AS 분획은 mouse내의 특정

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Plant Regeneration from Floral Stem Cultures of Nymphoides indica (L.) O. Kuntze. via Somatic Embryogenesis (어리연꽃 (Nymphoides indica (L.) O. Kuntze) 화경 배양으로부터 체세포배발생을 통한 식물체 재생)

  • Oh, Myung-Jin;Min, Sung-Ran;Liu, Jang-Ryol;Kim, Suk-Weon
    • Journal of Plant Biotechnology
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    • v.34 no.1
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    • pp.7-10
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    • 2007
  • Plant regeneration system from floral stem of Mymphoides indica via somatic embryogenesis was established. After four weeks of culture onto 1/2MS medium containing 2,4-D, pale-yellow globular structures and calluses were formed on the cut surface of floral stem explants. Upon transfer to 1/2MS basal medium, pale-yellow globular structures were developed into somatic embryos and normal plantlets. These results indicated that pale-yellow globular structures and calluses from floral stem were globular embryos and embryogenic calluses, respectively. The frequency of embryogenic callus formation from floral stem was reached to nearly 100% when floral stem was cultured onto 1/2Ms medium supplemented with low concentration of 2,4-D (0.1 to 0.3 mg/L). However, the higher concentration of 2,4-D resulted in decrease of the frequency of embryogenic callus formation. In this study, low concentration of 2,4-D had a stimulative role in embryogenic callus formation, whereas BA showed inhibitory role in callus formation. In comparison to floral stem, leaf explants showed low frequency of embryogenic callus formation. The highest frequency of embryogenic callus formation from leaf explants was 9.5% when leaf explants were cultured onto 1/2MS medium supplemented with 0.3 mg/L of 2,4-D. The plant regeneration system of Nymphoides indica established in this study, might be applied to mass proliferation, conservation of genetic resources and genetic transformation for molecular breeding.

Inhibitory Effect of LPS-Induced Plasminogen Activator Inhibitor-1 by Ascofuranone in Rat Kidney Fibroblast Cells (Ascofuranone에 의한 plasminogen activator inhibitor-1 발현저해 효과)

  • Chang, Young-Chae
    • Journal of Life Science
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    • v.19 no.10
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    • pp.1438-1443
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    • 2009
  • Renal fibrosis is a final common manifestation of every type of chronic kidney disease. Plasminogen activator inhibitor (PAI)-1 is induced by lipopolysaccharide (LPS) and is known to play an essential role in the progress of renal fibrosis. In this paper, we found that an isoprenoid antibiotic, ascofuranone (AF), suppresses expression of profibrotic factors, PAI-1 and promoter activity of PAI-1 induced by LPS in rat kidney fibroblast cells. We therefore investigated signaling pathway mediated inhibitory effects of LPS-induced PAI-1 by AF in rNRK-49F cells. PAI-1 expression is suppressed by treatment with kinase inhibitors for MEK-1/2, as it isin inhibition of PAI-1 expression by AF, and AF inhibits phosphorylation of ERK-1/2. This study suggest that AF suppresses expression of PAI-1 through the inhibition of an ERK-1/2-dependent signal transduction pathway. The data indicates the possibility that AF can be used to prevent the development and progression of renal fibrosis.

The systematic consideration of leaf epidermal microstructure in the tribe Forsythieae and its related genera (Oleaceae) (개나리족 및 근연분류군(물푸레나무과)의 잎 표피 미세구조에 대한 분류학적 검토)

  • Song, Jun-Ho;Hong, Suk-Pyo
    • Korean Journal of Plant Taxonomy
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    • v.43 no.2
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    • pp.118-127
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    • 2013
  • A comparative study of leaf epidermal microstructure in tribes Forsythieae (Abeliophyllum: 1 species, Forsythia: 12 taxa - 10 species, 2 varieties), Fontanesieae (Fontanesia: 2 species) including one related genus Myxopyrum belonging to Myxopyreae (Myxopyrum: 5 taxa - 4 species, 1 subspecies) was carried out using scanning electron microscopy (SEM) in order to evaluate their significance in the taxonomy. The leaves of investigated taxa are either hypostomatic or amphistomatic, but former is more frequent. The size range of the guard cells is 17.14-47.58 ${\times}$ 11.59- 44.25 ${\mu}m$: the smallest one was found in Forsythia giraldiana (17.48-22.96 ${\times}$ 11.64-12.88 ${\mu}m$), while the largest one was measured to Myxopyrum pierrei (31.50-41.75 ${\times}$ 32.53-44.25 ${\mu}m$). Anomocytic stomatal complex are most frequent type (rarely paracytic), usually both anomocytic and anisocytic occur in one leaf. In surface view both adaxial and abaxial anticlinal walls of the subsidiary cells are variable (e.g., straight/curved, undulate, sinuate, undulate/sinuate). Three types (simple unicellular and multicellular non-glandular, subsessile glandular) of trichomes are found in leaves. Finally, the systematic significance of the leaf epidermal micromorphological characters in idenfitication and elucidation of these tribe, between or within the genera including among the species is also briefly discussed.

DNA microarray analysis of RNAi plant regulated expression of NtROS2a gene encoding cytosine DNA demethylation (시토신 탈메틸화 관련 NtROS2a 유전자 발현을 제어한 RNAi 식물의 DNA microarray 분석)

  • Choi, Jang Sun;Lee, In Hye;Jung, Yu Jin;Kang, Kwon Kyoo
    • Journal of Plant Biotechnology
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    • v.43 no.2
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    • pp.231-239
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    • 2016
  • To study the transcript levels of epigenetically regulated genes in tobacco, we have developed a transgenic line OX1 overexpressing NtROS2a gene encoding cytosine DNA demethylation and a RNAi plant line RNAi13. It has been reported that salt- and $H_2O_2$-stress tolerance of these transgenic lines are enhanced with various phenotypic characters (Lee et al. 2015). In this paper, we conducted microarray analysis with Agilent Tobacco 4 x 44K oligo chip by using overexpression line OX1, RNAi plant line RNAi 13, and wild type plant WT. Differentially expressed genes (DEGs) related to metabolism, nutrient supply, and various stressed were up-regulated by approximately 1.5- to 80- fold. DEGs related to co-enzymes, metabolism, and methylation functional genes were down-regulated by approximately 0.03- to 0.7- fold. qRT-PCR analysis showed that the transcript levels of several candidate genes in OX1 and RNAi lines were significantly (p < 0.05) higher than those in WT, such as genes encoding KH domain-containing protein, MADS-box protein, and Zinc phosphodiesterase ELAC protein. On the other hand, several genes such as those encoding pentatricopeptide (PPR) repeat-containing protein, histone deacetylase HDAC3 protein, and protein kinase were decreased by approximately 0.4- to 1.0- fold. This study showed that NtROS2a gene encoding DNA glycosylase related to demethylation could regulate adaptive response of tobacco at transcriptional level.

Spinal Fusion Based on Ex Vivo Gene Therapy Using Recombinant Human BMP Adenoviruses (사람 골 형성 단백질 Ex vivo 유전자 치료법을 이용한 척추 유합)

  • Kim, Gi-Beom;Kim, Jae-Ryong;Ahn, Myun-Hwan;Seo, Jae-Sung
    • Journal of Yeungnam Medical Science
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    • v.24 no.2
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    • pp.262-274
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    • 2007
  • Purpose : Bone morphogenetic proteins (BMPs) play an important role in the formation of cartilage and bone, as well as regulating the growth of chondroblasts and osteoblasts. In this study, we investigated whether recombinant human BMP adenoviruses are available for ex vivo gene therapy, using human fibroblasts and human bone marrow stromal cells in an animal spinal fusion model. Materials and Methods : Human fibroblasts and human bone marrow stromal cells were transduced with recombinant BMP-2 adenovirus (AdBMP-2) or recombinant BMP-7 adenovirus (AdBMP-7), referred to as AdBMP-7/BMSC, AdBMP-2/BMSC, AdBMP-7/HuFb, and AdBMP-2/HuFb. We showed that each cell secreted active BMPs by alkaline phosphatase staining. Since AdBMP-2 or AdBMP-7 tranducing cells were injected into the paravertebral muscle of athymic nude mice, at 4 weeks and 7 weeks, we confirmed that new bone formation occurred by induction of spinal fusion on radiographs and histochemical staining. Results : In the region where the AdBMP-7/BMSC was injected, new bone formation was observed in all cases and spinal fusion was induced in two of these. AdBMP-2/BMSC induced bone formation and spinal fusion occurred among one of five. However, in the region where AdBMP/HuFb was injected, neither bone formation nor spinal fusion was observed. Conclusion : The osteoinductivity of AdBMP-7 was superior to that of AdBMP-2. In addition, the human bone marrow stromal cells were more efficient than the human fibroblasts for bone formation and spinal fusion. Therefore, the results of this study suggest that AdBMP-7/BMSC would be the most useful approach to ex vivo gene therapy for an animal spinal fusion model.

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Anti-inflammatory and Anti-allergic Effects of Herbal Extracts on Atopic Dermatitis ( Part II ) (약용식물 추출물의 아토피성 피부염에 대한 항염증 및 항알레르기 효과 (제 2 보))

  • Rang, Moon-Jeong
    • Journal of the Korean Applied Science and Technology
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    • v.30 no.1
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    • pp.173-182
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    • 2013
  • Atopic dermatitis is a chronic, relapsing inflammatory skin disease associated with dysfunction of skin barrier and cutaneous hyper-reactivity to environmental triggers. In the previous study, cytotoxicity, antioxidant, anti-inflammatory and anti-allergic activities were investigated for various herbal extracts such as Aloe vera L. (AV), Viola mandshurica W. Becker (VM), Punica granatum L. (PG), and Dendrobium nobile L. (DN) in order to develop effective therapeutic herbal extracts for atopic dermatitis, In this study, anti-inflammatory activities of these herb extracts in lipopolysaccharide (LPS)-induced macrophage RAW264.7 cells were further examined to find the underlying molecular mechanisms. The RT-PCR (reverse transcription polymerase chain reaction) analysis showed that PG, DN and AV inhibited effectively the gene expression of pro-inflammatory cytokines IL-6 and IL-$1{\beta}$ in LPS-stimulated macrophages, while VM did not. The transfection and luciferase analysis exhibited that all herbal extracts hindered the activation of transcription nuclear factor kappa B (NF-${\kappa}B$). The western blot analysis indicated that AV blocked the activation of only JNK MAP (c-Jun N-terminal kinase mitogen-activated protein) kinase not p38 MAP kinase, while VM, PG and DN did not show the activation of both JNK and p38 MAP kinases. These results suggest that AV, VM, PG, and DN have anti-inflammatory activities and thus have the potential to reduce and alleviate the symptoms of atopic dermatitis.

Overproduction and High Level Secretion of Glucose Oxidase in Saccharomyces cerevisiae (Glucose Oxidase의 Saccharomyces cerevisiae에서의 대량생산 및 고효율 분비)

  • 홍성용;최희경;이영호;백운화;정준기
    • Microbiology and Biotechnology Letters
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    • v.26 no.1
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    • pp.68-75
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    • 1998
  • The overproduction and high level secretion of Glucose Oxidase (GOD) from A. niger in S. cerevisiae was carried out by cloning GOD gene. For this purpose, using two different strong promoters (ADH1 promoter, GAL10 promoter) and signal sequences (${alpha}$-MF signal sequence of S. cerevisiae and ${alpha}$-amylase signal sequence of A. oryzae) and GAL7- and GOD terminator, four expression vectors were constructed. All the expression vectors were transformed in S. cerevisiae 2805 using auxotroph method. By the flask culture, transformants of pGAL expression vector series containing GAL 10 promotor showed much higher GOD productivity than transformants of pADH expression vector series containing ADH1 promoter Transformants of pGALGO2 containing GAL10 promotor and ${alpha}$-amylase signal sequence has shown the best productivity of GOD ($GOD_{total}$: 10.3 unit/mL, $GOD_{ex}$: 8.7 unit/mL) at 115 hr. This value was three fold higher than that of pGALGO1 containing GAL 10 promotor and ${alpha}$-MF signal sequence, even if the same promotor was involved. Through the ${alpha}$-amylase signal sequence of A. oryzae, GOD was secreted much more than the case of ${alpha}$-MF signal sequence from S. cerevisiae. These results suggest that signal sequence may play a important roles in not only the secretion but also the overproduction of foreign protein. Secretion rate of GOD in pGALGO1 and pGALGO2 was 89% and 84%, respectively, Because of the overglycosylation in S. cerevisiae the molecular weight of recombinant GOD in S. cerevisiae was much larger (250 kDa) than that of nature GOD in A. niger (170 kDa).

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