• 제목/요약/키워드: molecular analyses

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RAPD에 의한 한국산 붓꽃속(Iris)의 계통분류학적 연구 (A molecular systematic study of Korean Iris (Iridaceae) based on RAPD analysis)

  • 박선주;심정기;박홍덕
    • 식물분류학회지
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    • 제32권4호
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    • pp.383-396
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    • 2002
  • 한국산 붓꽃속(Iris L.; Iridaceae) 15종과 비교군인 등심붓꽃속(Sisyrinchium auguistifolium Mill.)과 범부채속(Belamcanda chinensis (L). DC.)을 포함한 17분류군을 대상으로 RAPD분석이 수행되었다. 10개의 oligoprimer를 이용한 효소중합반응에서 80개의 유효한 polymorphic band가 확인되었다. 유전적 상사도 지수에 의하여 bootstrap을 포함한 neighbour-joining 방법을 통하여 계통수를 평가하였다. 한국산 붓꽃속은 3아속(Limniris, Crossiris, Pardanthopsis) 또는 2속과 1아속으로 분류되어지고, 붓꽃아속(Limniris)은 6개의 분계조를 형성하고 있어 Waddick(1992)이 설정한 2절, 6계열의 분류체계가 더 타당하다고 생각된다. Chinensis계열은 금붓꽃계열과 각시붓꽃계열로 구분되어진다. Tripetalae계열에 속하는 부채붓꽃은 Sibiricae계열(시베리아붓꽃, 붓꽃)과 매우 가까운 유연관계를 형성하고 있으며, Ensatae계열에 속하는 타래붓꽃은 Ruthenicae계열에 속하는 솔붓꽃, 난장이붓꽃과 가까운 유연관계를 나타내고 있다. RAPD 분석자료를 기초로 한 한국산 붓꽃속의 계통학적 연구는 형태학적, ITS 염기서열 결과와 비슷한 결과를 제시하였고, 많은 학자들에 의해 논란이 된 분류군들의 문제를 해결하는데 유용한 접근방법이라 생각되며, 전체 붓꽃속 수준에서의 계통분석에 유용한 도구로 이용될 수 있는 것으로 사료된다.

Hypoxia Upregulates Mitotic Cyclins Which Contribute to the Multipotency of Human Mesenchymal Stem Cells by Expanding Proliferation Lifespan

  • Lee, Janet;Kim, Hyun-Soo;Kim, Su-Min;Kim, Dong-Ik;Lee, Chang-Woo
    • Molecules and Cells
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    • 제41권3호
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    • pp.207-213
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    • 2018
  • Hypoxic culture is widely recognized as a method to efficiently expand human mesenchymal stem cells (MSCs) without loss of stem cell properties. However, the molecular basis of how hypoxia priming benefits MSC expansion remains unclear. In this report, our systemic quantitative proteomic and RT-PCR analyses revealed the involvement of hypoxic conditioning activated genes in the signaling process of the mitotic cell cycle. Introduction of screened two mitotic cyclins, CCNA2 and CCNB1, significantly extended the proliferation lifespan of MSCs in normoxic condition. Our results provide important molecular evidence that multipotency of human MSCs by hypoxic conditioning is determined by the mitotic cell cycle duration. Thus, the activation of mitotic cyclins could be a potential strategy to the application of stem cell therapy.

High-concentration Epigallocatechin Gallate Treatment Causes Endoplasmic Reticulum Stress-mediated Cell Death in HepG2 Cells

  • Ahn, Joon-Ik;Jeong, Kyoung-Ji;Ko, Moon-Jeong;Shin, Hee-Jung;Chung, Hye-Joo;Jeong, Ho-Sang
    • Genomics & Informatics
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    • 제7권2호
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    • pp.97-106
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    • 2009
  • Epigallocatechin gallate (EGCG), a well-known antioxidant molecule, has been reported to cause hepatotoxicity when used in excess. However, the mechanism underlying EGCG-induced hepatotoxicity is still unclear. To better understand the mode of action of EGCG-induced hepatotoxicity, we examined the effect of EGCG on human hepatic gene expression in HepG2 cells using microarrays. Analyses of microarray data revealed more than 1300 differentially expressed genes with a variety of biological processes. Upregulated genes showed a primary involvement with protein-related biological processes, such as protein synthesis, protein modification, and protein trafficking, while downregulated genes demonstrated a strong association with lipid transport. Genes involved in cellular stress responses were highly upregulated by EGCG treatment, in particular genes involved in endoplasmic reticulum (ER) stress, such as GADD153, GADD34, and ATF3. In addition, changes in genes responsible for cholesterol synthesis and lipid transport were also observed, which explains the high accumulation of EGCG-induced lipids. We also identified other regulatory genes that might aid in clarifying the molecular mechanism underlying EGCG-induced hepatotoxicity.

Taxonomic Position and Affinities of Isopyrum mandshuricum within Korean Isopyroideae (Ranunculaceae) Based on Molecular Data

  • Lee, Nam-Sook;Yeau, Sung-Hee;Kim, Ji-Hyun;Kim, Min-Ju
    • Animal cells and systems
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    • 제3권2호
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    • pp.133-141
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    • 1999
  • To examine the taxonomic position and affinities of Isopyrum mandshuricum (Ranunculaceae) and related taxa, genetic analysis were carried out on the basis of isozyme patterns and ITS sequences. Molecular data, both isozyme patterns and ITS sequences suggest that I. mandshuricum is closely related to Enemion raddeanum than to Semiaquilegia adoxoides. The estimation of genetic identities by isozyme analysis reveals that I. manshuricum is genetically distant from E. raddeanum. The phylogenetic tree based on molecular data is rather congruent with the phenogram based on quantitative morphological characteristics, but not consistent with one based on qualitative morphological characteristics. Incongruencies between molecular and qualitative morphological data provide clues to re-evaluate several morphological features.

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Phylogenetic study of trichaptum inferred from nuclear ribosomal DNA sequences

  • Ko, Kwon-Soo;Hong, Soon-Gyu;Jung, Hack-Sung
    • Journal of Microbiology
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    • 제35권2호
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    • pp.79-86
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    • 1997
  • For the phylogenetic study of the genus Trichaptum, nuclear ribosomal DNA sequences from eight strains of four Trichaptium species were examined. Phylogenetic trees were constructed using molecular data on 18 rDNA and 5.8S rDNA and thei ITSs. Parsimony analyses of the Trichaptum species showed that T. biforme and T. laricinum made a monophyletic group respectively, suggesting that each species is phylogenetically independent. However, T. abietum represented a polyphyletic group and T. fusco-violaceum formed a polytomous group, suggesting that these species could be in the process of evolutionary differentiation. Examination of base substitutions of the 18S rRNA gene reveals that the C-T transition is most predominant and that there is a stronger transition bias between closely related organisms rather than between distantly related ones.

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MicroSPECT and MicroPET Imaging of Small Animals for Drug Development

  • Jang, Beom-Su
    • Toxicological Research
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    • 제29권1호
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    • pp.1-6
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    • 2013
  • The process of drug discovery and development requires substantial resources and time. The drug industry has tried to reduce costs by conducting appropriate animal studies together with molecular biological and genetic analyses. Basic science research has been limited to in vitro studies of cellular processes and ex vivo tissue examination using suitable animal models of disease. However, in the past two decades new technologies have been developed that permit the imaging of live animals using radiotracer emission, X-rays, magnetic resonance signals, fluorescence, and bioluminescence. The main objective of this review is to provide an overview of small animal molecular imaging, with a focus on nuclear imaging (single photon emission computed tomography and positron emission tomography). These technologies permit visualization of toxicodynamics as well as toxicity to specific organs by directly monitoring drug accumulation and assessing physiological and/or molecular alterations. Nuclear imaging technology has great potential for improving the efficiency of the drug development process.

Protein Phosphatases Involved in Regulating Mitosis: Facts and Hypotheses

  • Kim, Hyun-Soo;Fernandes, Gary;Lee, Chang-Woo
    • Molecules and Cells
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    • 제39권9호
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    • pp.654-662
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    • 2016
  • Almost all eukaryotic proteins are subject to post-translational modifications during mitosis and cell cycle, and in particular, reversible phosphorylation being a key event. The recent use of high-throughput experimental analyses has revealed that more than 70% of all eukaryotic proteins are regulated by phosphorylation; however, the mechanism of dephosphorylation, counteracting phosphorylation, is relatively unknown. Recent discoveries have shown that many of the protein phosphatases are involved in the temporal and spatial control of mitotic events, such as mitotic entry, mitotic spindle assembly, chromosome architecture changes and cohesion, and mitotic exit. This implies that certain phosphatases are tightly regulated for timely dephosphorylation of key mitotic phosphoproteins and are essential for control of various mitotic processes. This review describes the physiological and pathological roles of mitotic phosphatases, as well as the versatile role of various protein phosphatases in several mitotic events.

범용성 DNA 바코드 분석 기반 한국산 천남성속(Arisaema) 식물의 분자계통학적 연구 (Study on Molecular Phylogenetics of Korean Arisaema Species Based on Universal DNA Barcodes)

  • 노푸름;한경숙;김욱진;양선규;최고야;고성철;문병철
    • 한국자원식물학회지
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    • 제31권1호
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    • pp.37-51
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    • 2018
  • 국내에 분포하는 천남성속의 계통학적 유연관계와 한약재 천남성(Arisaematis Rhizoma)으로 사용되는 천남성 3종(둥근잎천남성, 두루미천남성, 일파산남성)에 대한 분류학적 특징을 분석하기 위하여 천남성속 식물에 대한 분자계통학적 연구를 수행하였다. 3개의 범용성 DNA 바코드(ITS, matK, rbcL) 염기서열을 이용하여 국내 분포 천남성속 8분류군과 중국에 분포하는 약전수재 종 1분류군을 포함하는 9종 50개 시료와 같은 과의 Dracunculus vulgaris를 군외군으로 하여 유연관계를 분석하였다. 3개의 개별 DNA 바코드 염기서열과 이들을 유합한 염기서열로 계통학적 유연관계를 분석한 결과, 천남성속의 9 분류군은 6개의 독립적인 분계조를 형성하며 구별되었으며(Clade I, 둥근잎천남성 및 천남성; Clade II, 점박이천남성 및 섬남성; Clade III, 큰천남성; Clade IV, 일파산남성; Clade V, 두루미천남성; Clade VI, 무늬천남성 및 거문천남성), 이들 6개의 분계조는 각각 Pedatisecta절, Sinarisaema절, 및 Tortuosa절로 분류되었다. 또한 이들 DNA 바코드 구간의 비교 결과는 천남성속 식물의 종 및 종 이하 분류 단위의 분류학적 재검토의 필요성에 대한 중요한 정보를 제공하였다. 하지만 대한민국약전에 수재되어 한약재로 사용가능한 3종의 천남성 기원종의 종내 분류학적 연관성이나 분자계통학적 특징은 확인되지 않았다.

Bacillus anthracis Spores Influence ATP Synthase Activity in Murine Macrophages

  • Seo, Gwi-Moon;Jung, Kyoung-Hwa;Kim, Seong-Joo;Kim, Ji-Cheon;Yoon, Jang-Won;Oh, Kwang-Keun;Lee, Jung-Ho;Chai, Young-Gyu
    • Journal of Microbiology and Biotechnology
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    • 제18권4호
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    • pp.778-783
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    • 2008
  • Anthrax is an infectious disease caused by toxigenic strains of the Gram-positive bacterium Bacillus anthracis. To identify the mitochondrial proteins that are expressed differently in murine macrophages infected with spores of B. anthracis Sterne, proteomic and MALDI-TOF/MS analyses of uninfected and infected macrophages were conducted. As a result, 13 mitochondrial proteins with different expression patterns were discovered in the infected murine macrophages, and some were identified as ATP5b, NIAP-5, ras-related GTP binding protein B isoform CRAa, along with several unnamed proteins. Among these proteins, ATP5b is related to energy production and cytoskeletal rearrangement, whereas NIAP-5 causes apoptosis of host cells due to binding with caspase-9. Therefore, this paper focused on ATP5b, which was found to be down regulated following infection. The downregulated ATP5b also reduced ATP production in the murine macrophages infected with B. anthracis spores. Consequently, this study represents the first mitochondrial proteome analysis of infected macrophages.

Formation of Flavone Di-O-Glucosides Using a Glycosyltransferase from Bacillus cereus

  • Ahn, Byoung-Chan;Kim, Bong-Gyu;Jeon, Young-Min;Lee, Eun-Jeong;Lim, Yoong-Ho;Ahn, Joong-Hoon
    • Journal of Microbiology and Biotechnology
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    • 제19권4호
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    • pp.387-390
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    • 2009
  • Microbial UDP-glycosyltransferases can convert many small lipophilic compounds into glycons using uridine-diphosphate-activated sugars. The glycosylation of flavonoids affects solubility, stability, and bioavailability. The gene encoding the UDP-glycosyltransferase from Bacillus cereus, BcGT-3, was cloned by PCR and sequenced. BcGT-3 was expressed in Escherichia coli BL21(DE3) with a glutathione S-transferase tag and purified using a glutathione S-transferase affinity column. BcGT-3 was tested for activity on several substrates including genistein, kaempferol, luteolin, naringenin, and quercetin. Flavonols were the best substrates for BcGT-3. The enzyme dominantly glycosylated the 3-hydroxyl group, but the 7-hydroxyl group was glycosylated when the 3-hydroxyl group was not available. The kaempferol reaction products were identified as kaempferol-3-O-glucoside and kaempferol-3,7-O-diglucoside. Kaempferol was the most effective substrate tested. Based on HPLC, LC/MS, and NMR analyses of the reaction products, we conclude that BcGT-3 can be used for the synthesis of kaempferol 3,7-O-diglucose.