• 제목/요약/키워드: Functional characterization

검색결과 793건 처리시간 0.029초

Analysis and characterization of the functional TGFβ receptors required for BMP6-induced osteogenic differentiation of mesenchymal progenitor cells

  • Zhang, Yan;Zhang, De-Ying;Zhao, Yan-Fang;Wang, Jin;He, Juan-Wen;Luo, Jinyong
    • BMB Reports
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    • 제46권2호
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    • pp.107-112
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    • 2013
  • Although BMP6 is highly capable of inducing osteogenic differentiation of mesenchymal progenitor cells (MPCs), the molecular mechanism involved remains to be fully elucidated. Using dominant negative (dn) mutant form of type I and type II $TGF{\beta}$ receptors, we demonstrated that three dn-type I receptors (dnALK2, dnALK3, dnALK6), and three dn-type II receptors (dnBMPRII, dnActRII, dnActRIIB), effectively diminished BMP6-induced osteogenic differentiation of MPCs. These findings suggested that ALK2, ALK3, ALK6, BMPRII, ActRII and ActRIIB are essential for BMP6-induced osteogenic differentiation of MPCs. However, MPCs in this study do not express ActRIIB. Moreover, RNA interference of ALK2, ALK3, ALK6, BMPRII and ActRII inhibited BMP6-induced osteogenic differentiation in MPCs. Our results strongly suggested that BMP6-induced osteogenic differentiation of MPCs is mediated by its functional $TGF{\beta}$ receptors including ALK2, ALK3, ALK6, BMPRII, and ActRII.

Functional Expression and Characterization of Recombinant NADPH-P450 Reductase from Malassezia globosa

  • Lee, Hwa-Youn;Park, Hyoung-Goo;Lim, Young-Ran;Lee, Im-Soon;Kim, Beom-Joon;Seong, Cheul-Hun;Chun, Young-Jin;Kim, Dong-Hak
    • Journal of Microbiology and Biotechnology
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    • 제22권1호
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    • pp.141-146
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    • 2012
  • Malassezia globosa is a common pathogenic fungus that causes skin diseases including dandruff and seborrheic dermatitis in humans. Analysis of its genome identified a gene (MGL_1677) coding for a putative NADPH-P450 reductase (NPR) to support the fungal cytochrome P450 enzymes. The heterologously expressed recombinant M. globosa NPR protein was purified, and its functional features were characterized. The purified protein generated a single band on SDS-PAGE at 80.74 kDa and had an absorption maximum at 452 nm, indicating its possible function as an oxidized flavin cofactor. It evidenced NADPH-dependent reducing activity for cytochrome c or nitroblue tetrazolium. Human P450 1A2 and 2A6 were able to successfully catalyze the O-deethylation of 7-ethoxyresorufin and the 7-hydroxylation of coumarin, respectively, with the support of the purified NPR. These results demonstrate that purified NPR is an orthologous reductase protein that supports cytochrome P450 enzymes in M. globosa.

Invertebrate Models Used for Characterization of Drug Dependence and Development of Anti-Drug Dependent Agents

  • Chang Hyun-Sook;Kim Ha-Won;Lee Dong-Hee
    • Biomolecules & Therapeutics
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    • 제14권1호
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    • pp.1-10
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    • 2006
  • Drug dependence deals a heavy socioeconomic burden to the society. For adolescents, the damage from drug dependence is greater than adults considering their higher susceptibility to drug effect and increasing chance for violence leading to criminal punishment process. Habitual drug use depends on genetic and environmental factors and the complex interactions between the two. Mammalian model systems have been useful in understanding the neurochemical and cellular impacts of abused drugs on specific regions of the brain, and in identifying the molecular targets of drugs. More elucidation is required whether biological effects of drugs actually cause the habitual dependence at the cellular level. Although there is much insight available on the nature of drug abuse problems, none of the systems designed to help drug dependent individuals is efficient in screening functional ingredients of the drug, and thus resulting in the failure of helping drug dependent individuals recover from drug dependence. Alternative model systems draw the attention of researchers, such as the invertebrate model systems of nematodes (Caenorhabditis elegans) and fruit flies (Drosophila melanogaster). These models should provide new insight into the mechanisms leading to the behavior of drug users (even functional studies analyzing molecular mechanism), and screening useful components to help remove drug dependence among drug users. The relatively simple anatomy and gene expression of the invertebrate model systems should enable researchers to coordinate current knowledge on drug abuse. Furthermore, the invertebrate model systems should facilitate advance in experiments on the susceptibility of specific genetic backgrounds and the interaction between genetic factors to drug dependence.

Characterization of EST Gene in the Bovine Corpus Luteum during the Estrous Cycle

  • Lee, Eunyoung;Kim, Sang Hwan;Kim, Byung-Gak;Yoon, Jong Taek
    • 한국발생생물학회지:발생과생식
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    • 제19권4호
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    • pp.227-234
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    • 2015
  • The objective of this study was to investigate the expression of bovine luteum expressed sequence tags (ESTs), vascular endothelial growth factor (VEGF), and tumor necrosis factor receptor 1 (TNFR1) and the presence of functional ESTs in the bovine corpus luteum (CL) during different stages of the estrus cycle. Reverse transcription-polymerase chain reaction (RT-PCR) analysis showed a difference in the expression of ESTs during the CL stage. Concentration of ESTs in the CL tissue increased significantly from the mid-luteal stage and decreased thereafter. RT-PCR analysis showed higher levels of the EST genes in the CL of the mid-luteal stage than in other stages, and the same level of expression of VEGF. Immunohistochemistry analysis of the tissue from CL formation to regression showed low cytosol and aggregation of the nucleus. And activity caspase 3 (apoptosis detector) was most strongly detected in the CL1 stage of bovine. During the estrous cycle, the cytosol was magnified and differentiation of the nucleus was clearly manifested. The ESTs affected the CL, and the relationship between VEGF and TNFR1 played a pivotal role for CL development and activation, dependent on the stage of CL. These results suggest local production of ESTs, the presence of functional ESTs in the bovine CL, and that ESTs play a role in regulating the function of cell death in bovine CL.

Development and Validation of Single Nucleotide Polymorphism (SNP) Markers from an Expressed Sequence Tag (EST) Database in Olive Flounder (Paralichthys olivaceus)

  • Kim, Jung Eun;Lee, Young Mee;Lee, Jeong-Ho;Noh, Jae Koo;Kim, Hyun Chul;Park, Choul-Ji;Park, Jong-Won;Kim, Kyung-Kil
    • 한국발생생물학회지:발생과생식
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    • 제18권4호
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    • pp.275-286
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    • 2014
  • To successful molecular breeding, identification and functional characterization of breeding related genes and development of molecular breeding techniques using DNA markers are essential. Although the development of a useful marker is difficult in the aspect of time, cost and effort, many markers are being developed to be used in molecular breeding and developed markers have been used in many fields. Single nucleotide polymorphisms (SNPs) markers were widely used for genomic research and breeding, but has hardly been validated for screening functional genes in olive flounder. We identified single nucleotide polymorphisms (SNPs) from expressed sequence tag (EST) database in olive flounder; out of a total 4,327 ESTs, 693 contigs and 514 SNPs were detected in total EST, and these substitutions include 297 transitions and 217 transversions. As a result, 144 SNP markers were developed on the basis of 514 SNP to selection of useful gene region, and then applied to each of eight wild and culture olive flounder (total 16 samples). In our experimental result, only 32 markers had detected polymorphism in sample, also identified 21 transitions and 11 transversions, whereas indel was not detected in polymorphic SNPs. Heterozygosity of wild and cultured olive flounder using the 32 SNP markers is 0.34 and 0.29, respectively. In conclusion, we identified SNP and polymorphism in olive flounder using newly designed marker, it supports that developed markers are suitable for SNP detection and diversity analysis in olive flounder. The outcome of this study can be basic data for researches for immunity gene and characteristic with SNP.

Bifidobacterium lactis를 이용한 기능성 김치의 특성 (Characterization of Functional Kimchi Using Bifidobacterium lactis)

  • 김태운;박애경;김금란;이정민;정대균;김해영
    • 한국식품과학회지
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    • 제35권5호
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    • pp.924-927
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    • 2003
  • 비피더스 균주가 가지고 있는 여러 생리 활성을 나타내는 부가가치를 부여한 새로운 고기능성 심치 제품을 개발하기 위해 편성 혐기성 미생물인 비피더스균 가운데 산소에 내성이 있는 Bifidobacterium lactis를 선발하여 김치 제조에 적용하였다. 이 균주의 내산성과 내염성을 각각 알아본 결과 pH 30에서는 별다른 영향을 받지 않았으며 pH 2.5에서도 50% 이상의 생존율을 나타내었다. 그리고 NaCl 농도 3.5%에서도 사멸하지 않는 것으로 나타나 일반적인 김치의 pH와 염농도를 고려할 때 김치에 적용 가능한 것으로 판단되었다. 첨가된 Bifidobacterium lactis가 김치 내에서 생존함을 확인하기 위해서 PCR을 이용하여 확인한 결과 15일 이상 생존하고 있음을 확인할 수 있었다. 그리고 관능평가의 경우 Bifidobacterium lactis 첨가 김치가 전체적인 기호도에서 균주를 첨가하지 않은 대조군 김치와 유사한 특성을 나타냄으로써 Bifidobacterium lactis를 이용한 고기능성 김치의 제조가 가능할 것으로 판단되었다.

황과 산소를 함유하는 새로운 Schiff Base 고분자의 합성, 특성분석, 열적 안정성과 전도성 (Synthesis, Characterization, Thermal Stability and Conductivity of New Schiff Base Polymer Containing Sulfur and Oxygen Bridges)

  • Culhaoglu, Suleyman;Kaya, Ismet
    • 폴리머
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    • 제39권2호
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    • pp.225-234
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    • 2015
  • In this study, we proposed to synthesize thermally stable, soluble and conjugated Schiff base polymer (SbP). For this reason, a specific molecule namely 4,4'-thiodiphenol which has sulfur and oxygen bridge in its structure was used to synthesize bi-functional monomers. Bi-functional amino and carbonyl monomers namely 4,4'-[thio-bis(4,1-phenyleneoxy)] dianiline (DIA) and 4,4'-[thiobis(4,1-phenyleneoxy)]dibenzaldehyde (DIB) were prepared from the elimination reaction of 4,4'-thiodiphenol with 4-iodonitrobenzene and 4-iodobenzaldehyde, respectively. The structures of products were confirmed by elemental analysis, FTIR, $^1H$ NMR and $^{13}C$ NMR techniques. The molecular weight distribution parameters of SbP were determined by size exclusion chromatography (SEC). The synthesized SbP was characterized by solubility tests, TG-DTA and DSC. Also, conductivity values of SbP and SbP-iodine complex were determined from their solid conductivity measurements. The conductivity measurements of doped and undoped SbP were carried out by Keithley 2400 electrometer at room temperature and atmospheric pressure, which were calculated via four-point probe technique. When iodine was used as a doping agent, the conductivity of SbP was observed to be increased. Optical band gap ($E_g$) of SbP was also calculated by using UV-Vis spectroscopy. It should be stressed that SbP was a semiconductor which had a potential in electronic and optoelectronic applications, with fairly low band gap. SbP was found to be thermally stable up to $300^{\circ}C$. The char of SbP was observed 29.86% at $1000^{\circ}C$.

식초기반 샐러드 드레싱용 다시마 식초 제조 및 이화학적 특성 (Preparation and Physicochemical Characterization of Sea Tangle Vinegar for Utilization as Vinegar-Based Salad Dressing)

  • 한아름;서정희
    • 한국식품조리과학회지
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    • 제33권3호
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    • pp.300-306
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    • 2017
  • Purpose: This study aimed to prepare sea tangle vinegar and test its applicability as a vinegar-based functional salad dressing in terms of physicochemical properties. Methods: Sea tangle vinegar was prepared by mixing sea tangle with sugar and vinegar and fermenting the mixture at room temperature for 3 months. The resulting sea tangle vinegar was examined for its physicochemical properties and antioxidant activity with brewed vinegar and persimmon vinegar as controls. Results: The sea tangle vinegar showed significantly higher viscosity than control vinegars, and shear thinning behavior that is typical for salad dressing containing polymers. In addition, storage modulus (G′) of sea tangle vinegar was relatively high in dynamic viscosity measurement while that of control vinegars remained negligible. Together with the high soluble solids content of sea tangle vinegar, rheological behavior indicates that sea tangle vinegar had soluble polysaccharides extracted from sea tangle, consequently leading to an increase in viscosity. Titratable acidity (TA) and pH were 2.52% and 3.58, respectively, which satisfies the TA and pH requirements for microbiological safety of a salad dressing. Absorbance at 285 nm and Folin Ciocalteu's reagent method revealed that sea tangle vinegar contained antioxidative phenolic compounds. Conclusion: This study demonstrates that sea tangle vinegar could be potentially developed as a vinegar-based functional salad dressing when combined with sensory evaluation in the future.

Sol-gel 법을 이용한 코어-쉘 실리카-형광체의 제조 및 특성평가 (Synthesis and Characterization of Core-Shell Silica-Phosphor Nanoparticles via Sol-Gel Process)

  • 신원호;김세윤;정형모
    • 한국분말재료학회지
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    • 제25권1호
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    • pp.12-18
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    • 2018
  • Cost-effective functional phosphor nanoparticles are prepared by introducing low-cost $SiO_2$ spheres to rare-earth phosphor ($YVO_4:Eu^{3+}$, $YVO_4:Er^{3+}$, and $YVO_4:Nd^{3+}$) shells using a sol-gel synthetic method. These functional nanoparticles are characterized by X-ray diffraction, X-ray photoelectron spectroscopy, transmission electron microscopy, and general photoluminescence spectra. The $SiO_2$ sphere occupying the interior of the conventional phosphor is advantageous in significantly reducing the cost of expensive rare-earth phosphor nanoparticles. The sol-gel process facilitates the core-shell structure formation; the rare-earth shell phosphor has strong interactions with chelating agents on the surfaces of $SiO_2$ nanoparticles and thus forms layers of several nanometers in thickness. The photoluminescence wavelength is simply tuned by replacing the active materials of $Eu^{3+}$, $Er^{3+}$, and $Nd^{3+}$. Moreover, the photoluminescent properties of the core-shell nanoparticles can be optimized by manipulating the specific contents of active materials in the phosphors. Our simple approach substitutes low-cost $SiO_2$ for expensive rare-earth-based phosphor materials to realize cost-effective phosphor nanoparticles for various applications.

Isolation and Characterization of Endophytic Actinomycetes from Chinese Cabbage Roots as Antagonists to Plasmodiophora brassicae

  • Lee, Sun-Og;Choi, Gyung-Ja;Choi, Yong-Ho;Jang, Kyoung-Soo;Park, Dong-Jin;Kim, Chang-Jin;Kim, Jin-Cheol
    • Journal of Microbiology and Biotechnology
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    • 제18권11호
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    • pp.1741-1746
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    • 2008
  • This study was conducted to select endophytic actinomycetes as biocontrol agents against Chinese cabbage clubroot caused by Plasmodiophora brassicae. A total of 81 endophytic actinomycetes were isolated from surface-sterilized roots of Chinese cabbage that was grown on paddy field and upland soils collected from various locations in Korea. By using 16S ribosomal DNA (rDNA) gene sequencing, they were classified to 8 actinobacterial genera. The genus Microbispora (67%) was most frequently isolated, followed by Streptomyces (12%) and Micromonospora (11%). Three of the 81 isolates, when inoculated in germinated Chinese cabbage seeds and then transplanted to pots, effectively suppressed the occurrence of a post-inoculated strain of P. brassicae in the pots. They showed control values of 58% for strain A004, 33% for strain A011, and 42% for strain A018. Based on cell wall components, morphological characteristics, and phylogenetic analyses, the three antagonistic isolates were identified as Microbispora rosea subsp. rosea (A004 and A011) and Streptomyces olivochromogenes (A018). Further researches on the field efficacy and action modes of the three actinomycetes are in progress.