• 제목/요약/키워드: Non-Isolation

검색결과 549건 처리시간 0.034초

Isolation of a Natural Antioxidant, Dehydrozingerone from Zingiber officinale and Synthesis of lts Analogues for Recognition of Effective Antioxidant and Antityrosinase Agents

  • Kuo, Ping-Chung;Damu, Amooru G.;Cherng, Ching-Yuh;Jeng, Jye-Fu;Teng, Che-Ming;Lee, E-Jian;Wu, Tian-Shung
    • Archives of Pharmacal Research
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    • 제28권5호
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    • pp.518-528
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    • 2005
  • In the present study, the antioxidative and inhibitory activity of Zingiber officinale Rosc. Rhizomes-derived materials (on mushroom tyrosinase) were evaluated. The bioactive co mponents of Z. officinale rhizomes were characterized by spectroscopic analysis as zingerone and dehydrozingerone, which exhibited potent antioxidant and tyrosinase inhibition activities. A series of substituted dehydrozingerones [(E)-4-phenyl-3-buten-2-ones] were prepared in admirable yields by the reaction of appropriate benzaldehydes with acetone and the products were evaluated in terms of variation in the dehydrozingerone structure. The synthetic analogues were examined for their antioxidant and antityrosinase activities to probe the most potent analogue. Compound 26 inhibited Fe$^{2+}$-induced lipid peroxidation in rat brain homogenate with an IC$_{50}$ = 6.3${\pm}$0.4 ${\mu}$M. In the 1,1-diphenyl- 2-picrylhydrazyl (DPPH) radical quencher assay, compounds 2, 7, 17, 26, 28, and 29 showed radical scavenging activity equal to or higher than those of the standard antioxidants, like ${\alpha}$-tocopherol and ascorbic acid. Compound 27 displayed superior inhibition of tyrosinase activity relative to other examined analogues. Compounds 2, 17, and 26 exhibited non-competitive inhibition against oxidation of 3,4- dihydroxyphenylalanine (L-DOPA). From the present study, it was observed that both number and position of hydroxyl groups on aromatic ring and a double bond between C-3 and C-4 played a critical role in exerting the antioxidant and antityrosinase activity.

The transposition pattern of the Ac element and its use for targeted transposition in Arabidopsis thaliana

  • Machida, Yasunori;Onouchi, Hitoshi;Tanaka, Hirokazu;Hamada, Susumu;Ishikawa, Takaaki;Semiarti, Endang;Iwakawa, Hidekazu;Nomura, Kiyohito;Machida, Chiyoko
    • 한국식물학회:학술대회논문집
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    • 한국식물학회 1999년도 제13회 식물생명공학심포지움 New Approaches to Understand Gene Function in Plants and Application to Plant Biotechnology
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    • pp.11-15
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    • 1999
  • In order to evlauate feasibility of the gene tagging by the maize transposable element Ac in heterologous plant systems, we have investigated physical distances and directions of transposition of the element in Arabidopsis thaliana and tobacco cultured cell line BY-2. We prepared a T-DNA construct that carried a non-autonomous derivative of Ac with a site for cleavage by endonuclease I-Scel (designated dAc-I-RS element). Another cleavage site was also introduced into the T-DNA region outside dAc-I-RS. A number of transgenic Arabidopsis plants were generated, each of which had a single copy of the T-DNA at a different chromosomal location. To examine the pattern of transposition, three out of these transgenic plants were crossed with the Arabidopsis plant that carried the gene for Ac transposase and progeny in which dAc-I-RS had been transposed were isolated. After digestion of the genomic DNA of these progeny with I-SceI, sizes of segment of DNA were determined byd pulse-field gel electrophoresis. We also performed linkage analysis for the transposed elements and sites of mutations near the elements. Our results with three transgenic lines showed that 50% of all transposition events had occurred within 1,700 kilo-base pairs (kb) on the same chromosome, with 35% within 200 kb, and that the elements transposed in both directions on the chromosome with roughly equal probability. The data thus indicate that the Ac-Ds system is most useful for tagging of genes that are present within 200 kb of the chromosomal site of Ac in Arabidopsis. In addition, determination of the precise localization of the transposed dAc-I-RS element should definitely assist in map-based cloning of genes around insertion sites. In the present paper, we report typical examples of such gene isolation studies.

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세포내의 물의 상태

  • 강사욱
    • 한국식물학회:학술대회논문집
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    • 한국식물학회 1985년도 워크샵 및 심포지엄 북한산국립공원의 식생
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    • pp.51-57
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    • 1985
  • In order to evlauate feasibility of the gene tagging by the maize transposable element Ac in heterologous plant systems, we have investigated physical distances and directions of transposition of the element in Arabidopsis thaliana and tobacco cultured cell line BY-2. We prepared a T-DNA construct that carried a non-autonomous derivative of Ac with a site for cleavage by endonuclease I-Scel (designated dAc-I-RS element). Another cleavage site was also introduced into the T-DNA region outside dAc-I-RS. A number of transgenic Arabidopsis plants were generated, each of which had a single copy of the T-DNA at a different chromosomal location. To examine the pattern of transposition, three out of these transgenic plants were crossed with the Arabidopsis plant that carried the gene for Ac transposase and progeny in which dAc-I-RS had been transposed were isolated. After digestion of the genomic DNA of these progeny with I-SceI, sizes of segment of DNA were determined byd pulse-field gel electrophoresis. We also performed linkage analysis for the transposed elements and sites of mutations near the elements. Our results with three transgenic lines showed that 50% of all transposition events had occurred within 1, 700 kilo-base pairs (kb) on the same chromosome, with 35% within 200 kb, and that the elements transposed in both directions on the chromosome with roughly equal probability. The data thus indicate that the Ac-Ds system is most useful for tagging of genes that are present within 200 kb of the chromosomal site of Ac in Arabidopsis. In addition, determination of the precise localization of the transposed dAc-I-RS element should definitely assist in map-based cloning of genes around insertion sites. In the present paper, we report typical examples of such gene isolation studies.

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Derivation of Embryonic Germ Cells from Post Migratory Primordial Germ Cells, and Methylation Analysis of Their Imprinted Genes by Bisulfite Genomic Sequencing

  • Shim, Sang Woo;Han, Dong Wook;Yang, Ji Hoon;Lee, Bo Yeon;Kim, Seung Bo;Shim, Hosup;Lee, Hoon Taek
    • Molecules and Cells
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    • 제25권3호
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    • pp.358-367
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    • 2008
  • The embryonic germ cell (EGCs) of mice is a kind of pluripotent stem cell that can be generated from pre- and post-migratory primordial germ cells (PGCs). Most previous studies on DNA methylation of EGCs were restricted to 12.5 days post coitum (dpc). This study was designed to establish and characterize murine EGC lines from migrated PGCs as late as 13.5 dpc and to estimate the degrees of methylation of their imprinted genes as well as of the non-imprinted locus, Oct4, using an accurate and quantitative method of measurement. We established five independent EGC lines from post migratory PGCs of 11.5-13.5 dpc from C57BL/6 ${\times}$ DBA/2 F1 hybrid mouse fetuses. All the EGCs exhibited the typical features of pluripotent cells including hypomethylation of the Oct4 regulatory region. We examined the methylation status of three imprinted genes; Igf2, Igf2r and H19 in the five EGC lines using bisulfite genomic sequencing analysis. Igf2r was almost unmethylated in all the EGC lines irrespective of the their sex and stage of isolation; Igf2 and H19 were more methylated than Igf2r, especially in male EGCs. Moreover, EGCs derived at 13.5 dpc exhibited higher levels of DNA methylation than those from earlier stages. These results suggest that in vitro derived EGCs acquire different epigenotypes from their parental in vivo migratory PGCs, and that sex-specific de novo methylation occurs in the Igf2 and H19 genes of EGCs.

Isolation of Surfactant-Resistant Pseudomonads from the Estuarine Surface Microlayer

  • Louvado, Antonio;Coelho, Francisco J.R.C.;Domingues, Patricia;Santos, Ana L.;Gomes, Newton C.M.;Almeida, Adelaide;Cunha, Angela
    • Journal of Microbiology and Biotechnology
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    • 제22권3호
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    • pp.283-291
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    • 2012
  • Bioremediation efforts often rely on the application of surfactants to enhance hydrocarbon bioavailability. However, synthetic surfactants can sometimes be toxic to degrading microorganisms, thus reducing the clearance rate of the pollutant. Therefore, surfactant-resistant bacteria can be an important tool for bioremediation efforts of hydrophobic pollutants, circumventing the toxicity of synthetic surfactants that often delay microbial bioremediation of these contaminants. In this study, we screened a natural surfactant-rich compartment, the estuarine surface microlayer (SML), for cultivable surfactant-resistant bacteria using selective cultures of sodium dodecyl sulfate (SDS) and cetyl trimethylammonium bromide (CTAB). Resistance to surfactants was evaluated by colony counts in solid media amended with critical micelle concentrations (CMC) of either surfactants, in comparison with non-amended controls. Selective cultures for surfactant-resistant bacteria were prepared in mineral medium also containing CMC concentrations of either CTAB or SDS. The surfactantresistant isolates obtained were tested by PCR for the Pseudomonas genus marker gacA gene and for the naphthalene-dioxygenase-encoding gene ndo. Isolates were also screened for biosurfactant production by the atomized oil assay. A high proportion of culturable bacterioneuston was tolerant to CMC concentrations of SDS or CTAB. The gacA-targeted PCR revealed that 64% of the isolates were Pseudomonads. Biosurfactant production in solid medium was detected in 9.4% of tested isolates, all affiliated with genus Pseudomonas. This study shows that the SML is a potential source of surfactant-resistant and biosurfactant-producing bacteria in which Pseudomonads emerge as a relevant group.

유도적 돌연변이 유발 방법을 통한 1-ethyl-3-methylimidazolium acetate에 대해 내성을 갖는 돌연변이 효모 선별 (Isolation of Mutant Yeast Strains having Resistance to 1-ethyl-3-methylimidazolium Acetate through a Directed Evolutionary Approach)

  • 이유진;권덕호;박재범;하석진
    • 한국미생물·생명공학회지
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    • 제45권1호
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    • pp.51-56
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    • 2017
  • 목질계 바이오 매스 전처리에 사용되는 ionic liquid는 전처리 후 100% 회수되지 않아 잔존하는 ionic liquid의 독성이 직접적으로 미생물 균주의 생육에 나쁜 영향을 미쳐 에탄올 발효의 수율 및 생산성을 저해하는 문제를 가지고 있다. 본 연구에서는 ionic liquid에 저해를 받지 않으며 높은 ethanol 생산 효율을 가진 균주를 얻고자 유도적 돌연변이 유발 실험을 진행하였다. 선별된 돌연변이 균주 D452-B2와 D452-S3는 3% [EMIM][Ac]가 포함된 배지에서 glucose 소비속도는 $4.5g{\cdot}l^{-1}{\cdot}h^{-1}$$4.4g{\cdot}l^{-1}{\cdot}h^{-1}$로 모균주인 S. cerevisiae D452-2 균주에 비해 6배 가량 증가하였으며, ethanol 생산성은 각각 $1.99g{\cdot}l^{-1}{\cdot}h^{-1}$$2.0g{\cdot}l^{-1}{\cdot}h^{-1}$로 27배 가량 증가하였다.

소와 돼지도체에서 Yersinia enterocolitica의 분리 및 특성 (Characteristics of Yersinia enterocolitica isolates from beef and pork carcass)

  • 채희선;김주영;김지은;양윤모;진경선;신방우;김선홍;이정학
    • 한국동물위생학회지
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    • 제31권2호
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    • pp.195-205
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    • 2008
  • Yersinia enterocolitica is a zoonotic agent, and to cause food poisoning. This study was carried out to get some basic information for the control of Yersinia infection. A total of 1,680 samples were collected from beef and pork carcasses from January 2006 to December 2007 in Seoul. The isolation rate was higher in pork carcass than in beef carcass. Five (0.59%) Yersinia enterocolitica were isolated from the 840 of beef carcasses, and eighteen(2.14%) were isolated from the 840 of pork carcasses. Among 23 strains, 22 were classified into biotype 1A, and one was biotype 6. In serotyping of Y enterocolitica isolates, 21 strains were untypable (UT), and 2 were O5 and O8 respectively. In PCR, Ail gene was not detected in all of 23 strains that determined non-pathogenic. In antimicrobial susceptibility test, twelve strains (52.2%) of 23 isolates showed the multi -resistant patterns with over 3 drugs. PFGE was performed after the genomic DNA of twenty three isolates, which was digested with Xba I. the 23 isolates showed 12 ($A{\sim}L$) PFGE type.

Isolation and Characterization of a Theta Glutathione S-transferase Gene from Panax ginseng Meyer

  • Kim, Yu-Jin;Lee, Ok-Ran;Lee, Sung-Young;Kim, Kyung-Tack;Yang, Deok-Chun
    • Journal of Ginseng Research
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    • 제36권4호
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    • pp.449-460
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    • 2012
  • Plants have versatile detoxification systems to encounter the phytotoxicity of the wide range of natural and synthetic compounds present in the environment. Glutathione S-transferase (GST) is an enzyme that detoxifies natural and exogenous toxic compounds by conjugation with glutathione (GSH). Recently, several roles of GST giving stress tolerance in plants have demonstrated, but little is known about the role of ginseng GSTs. Therefore, this work aimed to provide further information on the GST gene present in Panax ginseng genome as well as its expression and function. A GST cDNA (PgGST) was isolated from P. ginseng cDNA library, and it showed the amino acid sequence similarity with theta type of GSTs. PgGST in ginseng plant was induced by exposure to metals, plant hormone, heavy metals, and high light irradiance. To improve the resistance against environmental stresses, full-length cDNA of PgGST was introduced into Nicotiana tabacum. Overexpression of PgGST led to twofold increase in GST-specific activity compared to the non-transgenic plants, and the GST overexpressed plant showed resistance against herbicide phosphinothricin. The results suggested that the PgGST isolated from ginseng might have a role in the protection mechanism against toxic materials such as heavy metals and herbicides.

모형 객체 패턴을 이용한 Federation 통합시험 방법 (A Method of Integration Testing for Federation using Mock Object Patterns)

  • 심준용;이용헌;이승영;김세환
    • 한국시뮬레이션학회논문지
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    • 제20권4호
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    • pp.41-48
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    • 2011
  • 개발 기능에 대한 단위시험 모듈을 구현할 경우 도메인 구현부와 시험 구현부의 종속성이 높기 때문에 단위시험 모듈의 재사용이 어렵다. 특히, 동일한 구조나 기반 프레임워크를 재사용하는 시스템의 경우 구성 소프트웨어의 내부 인터페이스를 위한 단위시험 모듈의 중복이 불가피하며, 통합 시험 코드는 해당 모듈 간 연동 인터페이스 구현에 종속되기 때문에 각 모듈의 개발 일정에 따라 단위시험 수행이 제한될 수 있다. 이러한 문제를 해결하기 위해서 TDD 기법 중 하나인 모형 객체(Mock Objects) 패턴을 이용한 단위시험 방법이 제안되었다. 이 방법은 도메인 모듈과 시험 모듈을 분리할 수 있도록 도메인 모듈을 대리하는 모형 객체를 생성하고, 해당 모형 객체를 시험 모듈과 통합함으로써 단위시험 모듈의 구현을 용이하게 한다. 본 논문은 HLA 시뮬레이션 시스템 개발에 참여하는 Federate의 Federation 통합 및 연동 시험을 용이하게 하기 위해서 모형 객체를 적용한 모형 Federate를 설계하고, 모형 Federate의 구성 모듈을 위한 테스트 프레임워크를 제안한다. 제안 프레임워크는 RTI 서비스를 위한 시험 함수를 제공하며, 해당 함수들은 xUnit 패턴에 의해 자동화 된다.

Isolation, Characterization and Numerical Taxonomy of Novel Oxalate-oxidizing Bacteria

  • Sahin, Nurettin;Gokler, Isa;Tamer, Abdurrahman
    • Journal of Microbiology
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    • 제40권2호
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    • pp.109-118
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    • 2002
  • The present work is aimed at providing additional new pure cultures of oxalate utilizing bacteria and its preliminary characterization for further work in the field of oxalate-metabolism and taxonomic studies. The taxonomy of 14 mesophilic, aerobic oxalotrophic bacteria isolated by an enrichment culture technique from soils rhizosphers, and the juice of the petiole/stem tissue of plants was investigated. Isolates were characterized with 95 morphological, biochemical and physiological tests. Cellular lipid components and carotenoids of isolates were also studied as an aid to taxonomic characterization. All isolates were Gram-negative, oxidase and catalase positive and no growth factors were required. In addition to oxalates, some of the strains grow on methanol and/or formate. The taxonomic similarities among isolates, reference strains or previously reported oxalotrophic bacteria were analysed by using the Simple Matching (S/ sub SM/) and Jaccard (S$\_$J/) Coefficients. Clustering was performed by using the unweighted pair group method with arithmetic averages (UPGMA) algorithm. The oxalotrophic strains formed five major and two single-member clusters at the 70-86% similarity level. Based on the numerical taxonomy, isolates were separated into three phenotypic groups. Pink-pigmented strains belonged to Methylobacterium extorquens, yellow-pigmented strains were most similar to Pseudomonas sp. YOx and Xanthobacter autorophicus, and heterogeneous non-pigmented strains were closely related to genera Azospirillum, Ancylobacter, Burkholderia and Pseudomonas. New strains belonged to the genera Pseudomonas, Azospirillum and Ancylobacter that differ taxonomically from other known oxalate oxidizers were obtained. Numerical analysis indicated that some strains of the yellow-pigmented and nonpigmented clusters might represent new species.