• Title/Summary/Keyword: 분자 생물학적 분석

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Structural Organization of Calmodulin Gene and Expression in Transgenic

  • 최영주
    • Journal of Life Science
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    • v.4 no.2
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    • pp.50-59
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    • 1994
  • 신호전달과정의 연구는 calcium이 messenger로서 작용한다고 밝혀진 후로 식물에서 $Ca^{++}$ -messenger system에 대한 생화학적 및 분자생물학적 분야에서의 연구는 급속하게 발전하게 되었다. 식물세포에서 calcium 이온들의 많은 작용은 EF hand family로서 알려진 calcium binding protein에 의해서 조절된다. Calmodulin (CaM)은 highy conserve 되어 있으며, 4개의 calcium binding domain을 가진 ubiquitous한 단백질이다. 본 연구는 calmodulin 유전자의 발현에 미치는 calcium, EGTA, calcium ionophore 및 calmodulin antagonist의 영향과 또한 외부신호(light, wounding), chemical 및 auxin 등의 영향을 reporter화 유전자의 분석에 의해서 CaM유전자의 발현기작을 규명하고자 하였고, 또한 calmodulin 유전자의 organ-specific 발현 및 calmodulin의 새로운 생리적인 기능도 연구하고자 하였다.

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Phylogenetic relationships of medicinal mushroom Sparassis crispa strains using the rDNA-ITS and CAPS analysis (rDNA-ITS 및 CAPS 분석에 의한 꽃송이버섯 (Sparassis crispa) 수집균주의 계통분류학적 특성구분)

  • Cheong, Jong-Chun;Lee, Myung-Chul;Jhune, Chang-Sung;Lee, Chan-Jung;Shin, Pyeong-Gyun
    • Journal of Mushroom
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    • v.8 no.1
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    • pp.27-32
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    • 2010
  • This study was carried out to analyze the genetic relationships among 22 strains of Sparassis crispa, which were collected from various regions of worldwide. The cleaved amplified polymorphic sequence were obtained from the ribosomal DNA ITS regions of each strain. Based on the sequence analysis, the presence of five different groups were observed. Most strains shared the high nucleotide sequence similarity (about 90%) to each other, except only one strain, KACC50866. Nucleotide sequence similarity of KACC50866 was below 10% to other strains, indicating the genetic relatedness of strain KACC50866 was low compared to other strains. More works such as mitochondria genome analysis should help to determine the precise genetic diversity of S. crispa strains.

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Laser Captured Microdissection

  • 이경아
    • The Zoological Society Korea : Newsletter
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    • v.18 no.2
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    • pp.21-25
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    • 2001
  • 대부분의 조직은 여러 가지 세포가 모여서 이루어지기 때문에 그 중의 어떤 특정세포에서 발현하는 물질을 분석하려면 조직을 이루고 있는 각각의 세포를 분리해내야 한다. 이렇게 순수하게 세포를 분리해내는 기술 중의 하나가 Laser Captured Microdissection (LCM)이 다. LCM의 개발로 기존에 사용되던 방법에 비하여 빠르고 간편하면서, 매우 정확하게 원하는 세포를 순수 분리해서 그 세포의 분자생물학적 또는 생화학적인 분석을 할 수 있게 되었다. LCM은 현미경으로 조직절편을 관찰하면서 원하는 세포를 낮은 에너지의 laser를 사용하여 도려내는 방법으로 조직절편 이외에도 도말된 혈액이나 자궁경부 조직, 그리고 배양된 세포를 cytocentrifugation한 후에 원하는 세포를 포획 할 수도 있다. LCM을 이용한 연구는 여러 분야에서 다양하게 진행되고 있으며, 특히 같은 조직 내에 존재하는 정상세포와 전이중인 세포, 그리고 암세포를 구분해 냄으로써 암의 전이기전 및 병인 연구에 매우 큰 공헌을 하고 있다. 이렇게 분리된 세포는 RT-PCR, LOH (loss of heterozygosity), microsatellite instability, differential gene profiling, cDNA microarray, Western blot, 2D PAGE protein analysis 등의 기법을 접목하여 연구하게 된다. 본 논단을 통하여 1996년 개발된 LCM의 원리와 이제까지 LCM을 이용한 연구 성과를 살펴보고자 한다.

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Molecular Analysis and Enzymatic Characterization of Cathepsin B from Olive Flounder (Paralichthys olivaceus) (넙치 카텝신 B의 분자생물학적 분석 및 효소학적 특성 연구)

  • Jo, Hee-Sung;Kim, Na-Young;Lee, Hyung-Ho;Chung, Joon-Ki
    • Journal of Fisheries and Marine Sciences Education
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    • v.26 no.3
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    • pp.543-552
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    • 2014
  • Papain family중 하나인 cysteine protease는 근골격계 질환 치료를 위한target molecule로 인식 되어왔으며 Cathepsin B 는 단백질 분해의 초기과정에 관여하는 cysteine proteases 중 하나이다. 본 연구는 넙치의 cathepsin B 유전자의 발현 양상과 넙치 cathepsin B(PoCtB)의 클로닝, 발현 및 효소특성을 분석하였다. cDNA Library Screening을 이용하여 넙치의 cDNA를 클로닝하였다. 넙치의 동정된 cathepsin B 유전자는 993bp의 open reading frame과 330개의 아미노산으로 이루어져있다. Cathepsin B의 propeptide region 내에 GNFD motif와 occluding loop 가 존재함으로써 이것이 명백하게 cathepsin B group이라는 것을 보여주며, 계통 유전학적 분석 결과 다른 종의 cathepsin B에 비해 초창기에 분화되어 나온 것으로 사료된다. mature enzyme인 maPoCtB은 fusion protein인 glutathione S-transferase를 포함하는 pGEX-4T-1 vector에 삽입하여 E.coli 균주인 $DH5{\alpha}$ 내에 발현시켰다. 재조합 단백질인 PoCtB을 과발현 시킨 결과 53kDa의 분자량을 가진다. 넙치 cathepsin B 활성은 Z-Arg-Arg-AMC와 같은 fluorogenic 펩타이드 기질을 이용하여 측정되었고 적정 pH는 pH.7.5 이다.

Hypernetwork Classifiers for Microarray-Based miRNA Module Analysis (마이크로어레이 기반 miRNA 모듈 분석을 위한 하이퍼망 분류 기법)

  • Kim, Sun;Kim, Soo-Jin;Zhang, Byoung-Tak
    • Journal of KIISE:Software and Applications
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    • v.35 no.6
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    • pp.347-356
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    • 2008
  • High-throughput microarray is one of the most popular tools in molecular biology, and various computational methods have been developed for the microarray data analysis. While the computational methods easily extract significant features, it suffers from inferring modules of multiple co-regulated genes. Hypernetworhs are motivated by biological networks, which handle all elements based on their combinatorial processes. Hence, the hypernetworks can naturally analyze the biological effects of gene combinations. In this paper, we introduce a hypernetwork classifier for microRNA (miRNA) profile analysis based on microarray data. The hypernetwork classifier uses miRNA pairs as elements, and an evolutionary learning is performed to model the microarray profiles. miTNA modules are easily extracted from the hypernetworks, and users can directly evaluate if the miRNA modules are significant. For experimental results, the hypernetwork classifier showed 91.46% accuracy for miRNA expression profiles on multiple human canters, which outperformed other machine learning methods. The hypernetwork-based analysis showed that our approach could find biologically significant miRNA modules.

Confocal laser scanning microscopy image를 이용한 UASB granule의 메탄 생성 능력 측정

  • Lee, Yu-Jin;Kim, Hyo-Seop;An, Yeong-Hui;Park, Seong-Hun
    • 한국생물공학회:학술대회논문집
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    • 2000.04a
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    • pp.365-369
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    • 2000
  • Methanogenic activity of granular sludge was monitored by specific methanogenic activity (SMA) assay and confocal laser scanning microscopy (CLSM) during start-up of a thermophilic UASB reactor. Autofluorescence by CLSM could visualize the methanogenic bacterial population inside sludge granules and its intensity was proportional to SMA. Considering the complex procedures of SMA measurement, fluorescence quantification by CLSM can be suggested as a routine technique measuring methanogenic activity in UASB granules.

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Molecular phylogeny and the biogeographic origin of East Asian Isoëtes (Isoëtaceae) (동아시아 물부추속 식물의 분자계통 및 식물지리학적 기원에 대한 고찰)

  • CHOI, Hong-Keun;JUNG, Jongduk;NA, Hye-Ryun;KIM, Hojoon;KIM, Changkyun
    • Korean Journal of Plant Taxonomy
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    • v.48 no.4
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    • pp.249-259
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    • 2018
  • $Iso{\ddot{e}}tes$ L. ($Iso{\ddot{e}}taceae$) is a cosmopolitan genus of heterosporous lycopods containing ca. 200 species being found in lakes, streams, and wetlands of terrestrial habitats. Despite its ancient origin, worldwide distribution, and adaptation to diverse environment, species in $Iso{\ddot{e}}tes$ show remarkable morphological simplicity and convergence. Allopolyploidy appears to be a significant speciation process in the genus. These characteristics have made it difficult to assess the phylogenetic relationships and biogeographic history of $Iso{\ddot{e}}tes$ species. In recent years, these difficulties have somewhat been reduced by employing multiple molecular markers. Here, we reconstruct the phylogenetic relationships in East Asian $Iso{\ddot{e}}tes$ species. We also provide their divergence time and biogeographic origin using a fossil calibrated chronogram. East Asian $Iso{\ddot{e}}tes$ species are divided into two clades: I. asiatica and the remaining species. $Iso{\ddot{e}}tes$ asiatica from Hokkaido forms a clade with northeastern Russian and western North American $Iso{\ddot{e}}tes$ species. In clade I, western North America is the source area for the dispersal of $Iso{\ddot{e}}tes$ to Hokkaido and northeastern Russia via the Bering land bridge during the late Miocene. The remaining $Iso{\ddot{e}}tes$ species (I. sinensis, I. yunguiensis, I. hypsophila, I. orientalis, I. japonica, I. coreana, I. taiwanensis, I. jejuensis, I. hallasanensis) from East Asia form a sister group to Papua New Guinean and Australian species. The biogeographic reconstruction suggests an Australian origin for the East Asian species that arose through long-distance dispersal during the late Oligocene.

Prospect and Roles of Molecular Ecogenetic Techniques in the Ecophysiological Study of Cyanobacteria (남조류의 생리·생태 연구에서 분자생태유전학적 기법의 역할 및 전망)

  • Ahn, Chi-Yong
    • Korean Journal of Ecology and Environment
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    • v.51 no.1
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    • pp.16-28
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    • 2018
  • Although physiological and ecological characteristics of cyanobacteria have been studied extensively for decades, unknown areas still remain greater than the already known. Recently, the development of omics techniques based on molecular biology has made it possible to view the ecosystem from a new and holistic perspective. The molecular mechanism of toxin production is being widely investigated, by comparative genomics and the transcriptomic studies. Biological interaction between bacteria and cyanobacteria is also explored: how their interactions and genetic biodiversity change depending on seasons and environmental factors, and how these interactions finally affect each component of ecosystem. Bioinformatics techniques have combined with ecoinformatics and omics data, enabling us to understand the underlying complex mechanisms of ecosystems. Particularly omics started to provide a whole picture of biological responses, occurring from all layers of hierarchical processes from DNA to metabolites. The expectation is growing further that algal blooms could be controlled more effectively in the near future. And an important insight for the successful bloom control would come from a novel blueprint drawn by omics studies.

A comparison of the reproduction of two closely related species, tiger worm(Eisenia fetida) and red tiger worm(Eisenia andrei) when the organic sludge was suppied to them (유기성 슬러지 먹이에 대한 두 근연종인 줄지렁이(Eisenia fetida)와 붉은줄지렁이(Eisenia andrei)의 생식반응 비교)

  • Bae, Yoon-Hwan;Shin, Hyun-Gon
    • Journal of the Korea Organic Resources Recycling Association
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    • v.29 no.3
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    • pp.27-33
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    • 2021
  • CO I gene sequence analysis was applied to earthworms that had been used as test animals in toxicity test in Institute of Kyeongbook Agrochemicals and earthworms used as vermicomposting agents in the farm of Youngdong province to identify their species names. In terms of molecular species, the former was identified as Eisenia fetida and the latter was Eisenia andrei. Cocoons produced from Eisenia fetida was more than those from Eisenia andrei. And No. of adults developed from eggs of Eisenia fetida was more or less higher than those developed from eggs of Eisenia andrei. These results were contradictory to previous reports on two Eisenia spp.. When Eisenia fetida was crossed with Eisenia andrei, hybridized eggs were produced and adults were developed from those eggs, but cocoons and adults were much less than those from non-crossed Eisenia fetida or Eisenia andrei. This indicated that two Eisenia spp. were not distinctly different biological species because there was no complete 'reproductive isolation' between Eisenia fetida and Eisenia andrei. However, this also meant that Eisenia fetida and Eisenia andrei had already been on the tract of speciation.

Molecular Detection of Harmful Dinoflagellates (Dinophyceae) in Ballast Water (선박평형 수 내 유해 와편모조류(Dinophyceae)의 분자생물학적 검출)

  • Park, Tae-Gyu;Kim, Sung-Yeon
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.15 no.1
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    • pp.36-40
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    • 2010
  • Ballast water has been known as a major vector for global dispersal of toxic dinoflagellates and other microalgae. In this study, biodiversity in ships’ ballast water was examined using a dinoflagellate-oriented PCR primer set and species-specific real-time PCR. While motile dinoflagellates could be observe at very low cell densities by light microscopy,a wide range of dinoflagellate taxa including parasitic and phototrophic pico-dinoflagellates as well as harmful species to marine fish/shellfish was detected when techniques for cloning/sequencing of SSU rDNA of sample cells were used. Present result suggests that molecular methods including species-specific PCR primers may offer rapid and accurate detection of invasive species in ballast water.