• 제목/요약/키워드: Molecular Analysis

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Effects of Dark Treatment and DCMU on Desaturation of Galactolipids in Dunaliella salina (Dunaliella salina 당지질의 불포화반응에 미치는 암처리 및 DCMU의 효과)

  • 조성호
    • Journal of Plant Biology
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    • v.36 no.3
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    • pp.293-296
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    • 1993
  • Effects of dark treatment and N1-dichlorophenyl-N3-dimethylurea (DCMU) on the desaturation of galactolipids of Dunaliella salina were investigated to see whether light-driven photosynthetic electron transport is involved in in vivo desaturation of galactolipids. The incorporation of radioactive fatty acid precursors ([14C]lauric acid) into galactolipids, mainly composed of prokaryotic molecular species, was most affected among different polar lipid classes by both treatments. The analysis of specific radioactivities of individual galactolipid molecular species revealed that their synthesis was greatly inhibited by the treatments except for eukaryotic molecular species, 18 : 3/ 18 : 3 digalactosyldiacylglycerol, whose desaturation occurs in endoplasmic reticulum.

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CENSUS AND ANALYSIS OF GALACTIC MOLECULAR CLOUDS

  • HOJAEV, A.S.
    • Publications of The Korean Astronomical Society
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    • v.30 no.2
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    • pp.107-108
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    • 2015
  • In this project, all available databases of molecular and gas-dust clouds in the Galaxy were cross-identified by taking into account available properties, including position, angular dimensions, velocity, density, temperature and mass. An initial list of about 7000 entries was condensed into a cross-identified all-sky catalogue containing molecular and gas-dust clouds. Some relationships were studied between the main physical features of clouds. Finally, we prepared a complex observing program and address future work for filling in the gaps.

Surface-enhanced Raman Scattering on Chemically Prepared Silver Film for Trace Analysis

  • Boo, Doo-Wan;Kim, Kwan;Kim, Myung-Soo
    • Bulletin of the Korean Chemical Society
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    • v.7 no.1
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    • pp.55-58
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    • 1986
  • Analytical capability of surface-enhanced Raman scattering has been evaluated. Silver films prepared by homogeneous chemical reduction were used as the substrate. Detection limits for p-nitrobenzoic acid, thiophenol and rhodamine 6G were around $10{\sim}100\;pg$.

Combined Cytogenetic and Molecular Analyses for the Diagnosis of Prader-Willi/Angelman Syndromes

  • Borelina, Daniel;Engel, Nora;Esperante, Sebastian;Ferreiro, Veronica;Ferrer, Marcela;Torrado, Maria;Goldschmidt, Ernesto;Francipane, Liliana;Szijan, Irene
    • BMB Reports
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    • v.37 no.5
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    • pp.522-526
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    • 2004
  • Prader-Willi (PWS) and Angelman (AS) are syndromes of developmental impairment that result from the loss of expression of imprinted genes in the paternal (PWS) or maternal (AS) 15q11-q13 chromosome. Diagnosis on a clinical basis is difficult in newborns and young infants; thus, a suitable molecular test capable of revealing chromosomal abnormalities is required. We used a variety of cytogenetic and molecular approaches, such as, chromosome G banding, fluorescent in situ hybridization, a DNA methylation test, and a set of chromosome 15 DNA polymorphisms to characterize a cohort of 27 PWS patients and 24 suspected AS patients. Molecular analysis enabled the reliable diagnosis of 14 PWS and 7 AS patients, and their classification into four groups: (A) 6 of these 14 PWS subjects (44%) had deletions of paternal 15q11-q13; (B) 4 of the 7 AS patients had deletions of maternal 15q11-q13; (C) one PWS patient (8%) had a maternal uniparental disomy (UPD) of chromosome 15; (D) the remaining reliably diagnoses of 7 PWS and 3 AS cases showed abnormal methylation patterns of 15q11-q13 chromosome, but none of the alterations shown by the above groups, although they may have harbored deletions undetected by the markers used. This study highlights the importance of using a combination of cytogenetic and molecular tests for a reliable diagnosis of PWS or AS, and for the identification of genetic alterations.

Directed Mutagenesis of the Bacillus thuringiensis Cry11A Toxin Reveals a Crucial Role in Larvicidal Activity of Arginine-136 in Helix 4

  • Angsuthanasombat, Chanan;Keeratichamreon, Siriporn;Leetacheewa, Somphob;Katzenmeier, Gerd;Panyim, Sakol
    • BMB Reports
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    • v.34 no.5
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    • pp.402-407
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    • 2001
  • Based on the currently proposed toxicity model for the different Bacillus thuringiensis Cry $\delta$-endotoxins, their pore-forming activity involves the insertion of the ${\alpha}4-{\alpha}5$ helical hairpin into the membrane of the target midgut epithelial cell. In this study, a number of polar or charged residues in helix 4 within domain I of the 65-kDa dipteranactive Cry11A toxin, Lys-123, Tyr-125, Asn-128, Ser-130, Gln-135, Arg-136, Gln-139 and Glu-141, were initially substituted with alanine by using PCR-based directed mutagenesis. All mutant toxins were expressed as cytoplasmic inclusions in Escherichia coli upon induction with IPTG. Similar to the wild-type protoxin inclusion, the solubility of each mutant inclusion in the carbonate buffer, pH 9.0, was relatively low When E. coli cells, expressing each of the mutant proteins, were tested for toxicity against Aedes aegypti mosquito-larvae, toxicity was completely abolished for the alanine substitution of arginine at position 136. However, mutations at the other positions still retained a high level of larvicidal activity Interestingly, further analysis of this critical arginine residue by specific mutagenesis showed that conversions of arginine-136 to aspartate, glutamine, or even to the most conserved residue lysine, also abolished the wild-type activity The results of this study revealed an important determinant in toxin function for the positively charged side chain of arginine-136 in helix 4 of the Cry11A toxin.

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Integrative Analysis of Microarray Data with Gene Ontology to Select Perturbed Molecular Functions using Gene Ontology Functional Code

  • Kim, Chang-Sik;Choi, Ji-Won;Yoon, Suk-Joon
    • Genomics & Informatics
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    • v.7 no.2
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    • pp.122-130
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    • 2009
  • A systems biology approach for the identification of perturbed molecular functions is required to understand the complex progressive disease such as breast cancer. In this study, we analyze the microarray data with Gene Ontology terms of molecular functions to select perturbed molecular functional modules in breast cancer tissues based on the definition of Gene ontology Functional Code. The Gene Ontology is three structured vocabularies describing genes and its products in terms of their associated biological processes, cellular components and molecular functions. The Gene Ontology is hierarchically classified as a directed acyclic graph. However, it is difficult to visualize Gene Ontology as a directed tree since a Gene Ontology term may have more than one parent by providing multiple paths from the root. Therefore, we applied the definition of Gene Ontology codes by defining one or more GO code(s) to each GO term to visualize the hierarchical classification of GO terms as a network. The selected molecular functions could be considered as perturbed molecular functional modules that putatively contributes to the progression of disease. We evaluated the method by analyzing microarray dataset of breast cancer tissues; i.e., normal and invasive breast cancer tissues. Based on the integration approach, we selected several interesting perturbed molecular functions that are implicated in the progression of breast cancers. Moreover, these selected molecular functions include several known breast cancer-related genes. It is concluded from this study that the present strategy is capable of selecting perturbed molecular functions that putatively play roles in the progression of diseases and provides an improved interpretability of GO terms based on the definition of Gene Ontology codes.

High Level of Soluble Expression in Escherichia coli and Characterisation of the Cloned Bacillus thuringiensis Cry4Ba Domain III Fragment

  • Chayaratanasin, Poramed;Moonsom, Seangdeun;Sakdee, Somsri;Chaisri, Urai;Katzenmeier, Gerd;Angsuthanasombat, Chanan
    • BMB Reports
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    • v.40 no.1
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    • pp.58-64
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    • 2007
  • Similar to the other known structures of Bacillus thuringiensis Cry $\delta$-endotoxins, the crystal structure of the 65-kDa activated Cry4Ba toxin comprises three domains which are, from the N- to C-terminus, a bundle of $\alpha$-helices, a three-$\beta$-sheet domain, and a $\beta$-sandwich. To investigate the properties of the C-terminal domain III in isolation from the rest of the toxin, the cloned Cry4Ba-domain III was over-expressed as a 21-kDa soluble protein in Escherichia coli, which cross-reacted with anti-Cry4Ba domain III monoclonal antibody. A highly-purified domain III was obtained in a monomeric form by ion-exchange and size-exclusion FPLC. Circular dichroism spectroscopy indicated that the isolated domain III fragment distinctly exists as a $\beta$-sheet structure, corresponding to the domain III structure embodied in the Cry4Ba crystal structure. In vitro binding analysis via immuno-histochemical assay revealed that the Cry4Ba-domain III protein was able to bind to the apical microvilli of the susceptible Stegomyia aegypti larval midguts, albeit at lower-binding activity when compared with the full-length active toxin. These results demonstrate for the first time that the C-terminal domain III of the Cry4Ba mosquito-larvicidal protein, which can be isolated as a native folded monomer, conceivably participates in toxin-receptor recognition.