• Title/Summary/Keyword: Molecular Characteristics

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Mode of Action on EcoRI Restriction Endonuclease: EcoRI and EcoRI Variant N199H have Active Monomeric Forms

  • Kim, Jae-Jong;Koh, Suk-Hoon;Kim, Joong-Su;Lee, Dae-Sil
    • BMB Reports
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    • v.31 no.2
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    • pp.149-155
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    • 1998
  • The N199H variant of the EcoRI endonuclease has about twice the catalytic activity of the wild-type. A comparison of their biochemical characteristics, using synthetic oligonucleotides 5'-dAAAACTTAAGAAAAAAAAAAA-3' (KA) and 5'-dTTTTTGAATTCTTTTTTTTTT-3' (KT), helps to define the cleavage reaction pathway of these enzymes. Both EcoRI and EcoRI variant N199H were found to cleave singlestranded KA or KT about three times faster than the double-stranded forms, although the KT oligonucleotide was more susceptible. Using the ssDNA substrate in kinetic analyses, lower $K_m$ values were obtained for the N199H variant than for the wild-type at low (50 mM), as well as high (200 mM), sodium chloride concentrations. This difference between the endonucleases is attributed to a grealter accessibility for tbe substrate by the variant, and also a higher affinity for the DNA backbone. It also appears that the relative activities of the two enzymes, particularly at high ionic strength, are proportional to their populations in the monomeric enzyme form. That is, according to gel filtration data, half of the N199H molecules exist as monomers in 200 mM NaCl, whereas those of the wild-type are mainly dimeric. Consequently, the Asp199 residue of the EcoRI endonuclease may be implicated in the protein-protein interaction leading to dimerization, as well as in coupling to DNA substrates. In summary, it is proposed that active monomeric endonuclease molecules, derived from the dimeric enzyme, recognize and form a complex with a single stranded form of the DNA substrate, which then undergoes nucleophilic substitution and cleavage.

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Identification and Characterization of Osmotolerant Yeast Isolated from Soy Paste (된장에서 분리된 내염성 효모의 동정 및 특성조사)

  • Byun, Myung-Ok;Lee, Seung-Bum;Koo, Bon-Sung;Song, Jae-Kyeong;Ryu, Jin-Chang;Lee, Du-Hyung
    • The Korean Journal of Mycology
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    • v.27 no.3 s.90
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    • pp.181-186
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    • 1999
  • Osmotolerant yeast isolated from soy paste could grow on media with 2 M NaCl. This strain was identified as Zygosaccharomyces rouxii by biological characteristics, RFLP of ribosomal DNA and mating with compatible haploid strain. Growing rate of the Z. rouxii YDJ was slower than Saccharomyces cerevisiae. Z. rouxii YDJ accumulated trehalose, which is known as one of the osmolytic protectants, in cells cultured on media with salt. Enzyme activity of trehalose phosphate synthase related to trehalose biosynthesis of the YDJ was lower than those of S. cerevisiae. Trehalase activity related trehalose degradation was also lower in Z. rouxii YDJ than S. cerevisiae. However, as Z. rouxii accumulated trehalose by salt treatment, salt tolerancy of Z. rouxii was assumed to be related to trehalose in additon to glycerol.

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Molecular Identification of Adoxophyes honmai (Yasuda) (Lepidoptera: Tortricidae) Based on Mitochondrial COI Gene Sequences

  • Lee, So Young;Park, Hyungjin;Boo, Kyung Saeng;Park, Kyu-Tek;Cho, Soowon
    • Molecules and Cells
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    • v.19 no.3
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    • pp.391-397
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    • 2005
  • Molecular identification techniques are used where morphological characters are not useful for distinguishing species that resemble each other closely. The example studied here is the Adoxophyes species complex, in which A. orana (Fischer von $R{\ddot{o}}sslerstamm$) is officially the only known Korean species in the genus Adoxophyes (Lepidoptera: Tortricidae). However there have been suspicions that at least two types of A. orana exist in Korea based on the distribution and range of the host, with A. orana attacking apples and peaches, and another Adoxophyes sp. attacking tea and pears. The latter is presumed to be A. honmai (Yasuda), but the two have remained confused because of their extreme morphological similarity, despite several Asian studies of pheromonal and morphological characteristics. To confirm the occurrence of an Adoxophyes species other than A. orana in Korea, we compared 940 bp of the mitochondrial cytochrome oxidase I (COI) gene from 16 samples of Adoxophyes and found that there is a second Adoxophyes species different from A. orana. Comparison of the different sequences to that of Japanese A. honmai confirmed that they belong to the latter. From the sequence difference between the two Korean species, we were able to develop new PCR primer sets that distinguish them. This molecular identification technique with no enzyme digestion or sequencing step is a convenient and rapid way of differentiating between species that are hard to distinguish morphologically.

Synthesis of Highly Selective Polyimide Material and Comparison of Gas Permeability by Molecular Dynamics Study (고선택성 폴리이미드 소재의 합성 및 분자동력학 연구를 통한 기체투과도의 비교)

  • Lee, Jung Moo;Kim, Deuk Ju;Jeong, Moon Ki;Lee, Myung Gun;Park, Chi Hoon;Nam, Sang Yong
    • Membrane Journal
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    • v.25 no.2
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    • pp.162-170
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    • 2015
  • In this study, gas permeability of polyimide materials having a various amine group was measured and molecular dynamics was used to analyze the dynamic characteristics of the gas molecules in the polyimide by calculating the position and velocity of the gas molecules with change of the time. The gas permeability of polyimide membrane having substitution site which increase free volume in the polymer was increased. However, polyimide with rigid structure showed decreased gas permeability. As a result of analyzing the change in the gas permeation behavior using molecular dynamics simulations, we confirmed that the results show the same tendency with actual measurements of the gas permeability.

Biochemical Characterization and Genetic Diversity of Pongamia pinnata (L.) Pierre in Eastern India

  • Kumari, Kanchan;Sinha, Amrita;Singh, Sanjay;Divakara, B.N.
    • Journal of Forest and Environmental Science
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    • v.29 no.3
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    • pp.200-210
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    • 2013
  • Biochemical characteristics of 24 Pongamia pinnata genotypes (candidate plus trees) from three agroclimatic zones were estimated and molecular characterization through RAPD markers was done. Various biochemical characters viz. seed oil, total carbohydrates, protein, acid value and Iodine number recorded significant variation among different genotypes. The highest seed oil content was 41.87% while seeds of 14 genotypes recorded above average (32.11%) for the trait. Seed oil and protein content exhibited a significant positive correlation and moderate heritability. Out of the initially selected twenty-five random primers, twenty-two RAPD primers were found to be highly reproducible and produced a total of 183 loci of which 147 (80.32%) loci were polymorphic. Percentage of polymorphism varied from 44% to 100% with an average of 80.62%. High level of genetic variation was found among different genotypes of P. pinnata. Both molecular and oil content (biochemical) markers appeared useful in analyzing the extent of genetic diversity in Pongamia and the result of these analyses will help to better understand the genetic diversity and relationship among populations. Overall, the Pongamia genotypes included in the study showed a correlation with their geographical origins such that genotypes from the same region tend to have higher genetic similarity as compared to those from different regions. However, in UPGMA based Nei's analysis, some genotypes were found not to be grouped based on geographical origins possibly due to the exchange of germplasm over time between farmers across the regions. The results from oil content analyses showed that several genotypes in 'Central and Western Plateau' agroclimatic zone of Jharkhand displayed a good potential for high oil content. The study provides insight about P. pinnata populations in Jharkhand (India) and constitutes a set of useful background information that can be used as a basis for future breeding strategy and improvement of the species.

Construction of Microbial Fuel Cells Using Thermophilic Microorganisms, Bacillus licheniformis and Bacillus thermoglucosidasius

  • Choi, Young-Jin;Jung, Eun-Kyoung;Park, Hyun-Joo;Paik, Seung R.;Jung, Seun-Ho;Kim, Sung-Hyun
    • Bulletin of the Korean Chemical Society
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    • v.25 no.6
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    • pp.813-818
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    • 2004
  • A systematic study of microbial fuel cells comprised of thermophilic Bacillus licheniformis and Bacillus thermoglucosidasius has been carried out under various operating conditions. Substantial amount of electricity was generated when a redox mediator was used. Being affected by operation temperature, the maximum efficiency was obtained at 50$^{\circ}C$ with an open circuit voltage of ca. 0.7 V. While a small change around the optimum temperature did not make much effect on the cell performance, the rapid decrease in performance was observed above 70$^{\circ}C$. It was noticeable that fuel cell efficiency and discharge pattern strongly depended on the kind of carbon sources used in the initial culture medium. In the case of B. thermoglucosidasius, glucose alone was utilized constitutively as a substrate in the microbial fuel cell irrespective of used carbons sources. When B. licheniformis was cultivated with lactose as a carbon source, best charging characteristics were recorded. Trehalose, in particular, showed 41.2% coulombic efficiency when B. thermoglucosidasius was cultured in a starch-containing medium. Relatively good repetitive operation was possible with B. thermoglucosidasius cells up to 12 cycles using glucose as a carbon source, when they were cultured with lactose as an initial carbon source. This study demonstrates that highly efficient thermophilic microbial fuel cells can be constructed by a pertinent modulation of the operating conditions and by carefully selecting carbon sources used in the initial culture medium.

Phylogenetic study of the Genus Suaeda(Chenopodiaceae) based on chloroplast and nuclear DNA sequences from Korea (엽록체 DNA 및 핵 DNA 염기서열에 근거한 한국산 나문재속(명아주과)의 분류학적 연구)

  • Kim, Suk-Kyu;Chung, Sang Ok
    • Korean Journal of Environment and Ecology
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    • v.32 no.6
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    • pp.566-574
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    • 2018
  • The purpose of this study was to identify the phylogenetic relationships of the plants in the Korean genus Suaeda and to find out the molecular markers that could confirm the interspecies relationships in the family tree through molecular phylogenetic studies. We used the nuclear ribosomal DNA ITS and the chloroplast DNA matK, psbA-trnH, and trnL-trnF as the molecular markers. We could not distinguish between S. japonica and S. maritima and between S. maritima and S. australis in the ITS region and could not distinguish between S. japonica and S. australis with the base sequence in the psbA-trnH and trnL-trnF region. However, we analyzed the combinations of four molecular marker regions and confirmed that each of five plant species of the genus Suaeda formed the independent line. Therefore, it is considered that combinations of molecular markers would be useful for the analysis of phylogenetic relationships in the genus Suaeda. Further investigations of the ecological and morphological characteristics would be needed to understand the phylogenetic relationship and lineage diversification in the genus Suaeda.

Genetic overgrowth syndrome: A single center's experience

  • Cheon, Chong Kun;Kim, Yoo-Mi;Yoon, Ju Young;Kim, Young A
    • Journal of Genetic Medicine
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    • v.15 no.2
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    • pp.64-71
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    • 2018
  • Purpose: Overgrowth syndromes are conditions that involve generalized or localized areas of excess growth. In this study, the clinical, molecular, and genetic characteristics of Korean patients with overgrowth syndrome were analyzed. Materials and Methods: We recruited 13 patients who presented with overgrowth syndrome. All patients fulfilled inclusion criteria of overgrowth syndrome. Analysis of the clinical and molecular investigations of patients with overgrowth syndrome was performed retrospectively. Results: Among the 13 patients with overgrowth syndrome, 9 patients (69.2%) were found to have molecular and genetic causes. Among the seven patients with Sotos syndrome (SS), two had a 5q35microdeletion that was confirmed by fluorescent in situ hybridization. In two patients with SS, intragenic mutations including a novel mutation, c.5993T>A (p.M1998L), were found by Sanger sequencing. One patient had one copy deletion of NDS1 gene which was confirmed by multiplex ligation-dependent probe amplification. Among five patients with Beckwith-Wiedemann syndrome, three had aberrant imprinting control regions; 2 hypermethylation of the differentially methylated region of H19, 1 hypomethylation of the differentially methylated region of Kv. In one patient displaying overlapping clinical features of SS, a de novo heterozygous deletion in the chromosomal region 7q22.1-22.3 was found by single nucleotide polymorphism-based microarray. Conclusion: Considering high detection rate of molecular and genetic abnormalities in this study, rigorous investigations of overgrowth syndrome may be an important tool for the early diagnosis and genetic counseling. A detailed molecular analysis of the rearranged regions may supply the clues for the identification of genes involved in growth regulation.

The Structural Studies of Peptide P143 Derived from Apo B-100 by NMR

  • Lee, Ji-Eun;Kim, Gil-Hoon;Won, Ho-Shik
    • Journal of the Korean Magnetic Resonance Society
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    • v.25 no.4
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    • pp.58-63
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    • 2021
  • Apolipoprotein B-100 (apo B-100), the main protein component that makes up LDL (Low density lipoprotein), consists of 4,536 amino acids and serves to combine with the LDL receptor. The oxidized LDL peptides by malondialdehyde (MDA) or acetylation in vivo act as immunoglobulin (Ig) antigens and peptide groups were classified into 7 peptide groups with subsequent 20 amino acids (P1-P302). The biomimetic peptide P143 (IALDD AKINF NEKLS QLQTY) out of C-group peptides carrying the highest value of IgG antigens were selected for structural studies that may provide antigen specificity. Experimental results show that P143 has β-sheet in Ile[1]-Asn[9] and α-helice in Gln[16]-Tyr[20] structure. Homonuclear 2D-NMR (COSY, TOCSY, NOESY) experiments were carried out for NMR signal assignments and structure determination for P143. On the basis of these completely assigned NMR spectra and proton distance information, distance geometry (DG) and molecular dynamic (MD) were carried out to determine the structures of P143. The proposed structure was selected by comparisons between experimental NOE spectra and back-calculated 2D NOE results from determined structure showing acceptable agreement. The total Root-Mean-Square-Deviation (RMSD) value of P143 obtained upon superposition of all atoms were in the set range. The solution state P143 has a mixed structure of pseudo α-helix and β-turn(Phe[10] to Glu[12]). These results are well consistent with calculated structure from experimental data of NOE spectra. Structural studies based on NMR may contribute to the prevent oxidation studies of atherosclerosis and observed conformational characteristics of apo B-100 in LDL using monoclonal antibodies.

Strategies to Design Efficient Donor-Acceptor (D-A) Type Emitting Molecules: Molecular Symmetry and Electron Accepting Ability of D-A Type Molecules

  • Hyun Gi Kim;Young-Seok Baek;Sung Soo Kim;Sang Hyun Paek;Young Chul Kim
    • Applied Chemistry for Engineering
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    • v.34 no.6
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    • pp.633-639
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    • 2023
  • We synthesized 2-(10-methyl-10H-phenothiazin-3-yl)-5-phenyl-1,3,4-oxadiazole (MPPO) and 5,5-(10-methyl-10H-phenothiazin-3,7-diyl)-bis-(2-phenyl-1,3,4-oxadiazole) (DPPO). MPPO has both electron-donating and electron-accepting substituents with asymmetric molecular geometry. By incorporating one extra electron-accepting group into MPPO, we created a symmetric molecule, which is DPPO. The optical and electrochemical properties of these compounds were measured. The lowest unoccupied molecular orbital (LUMO) level of DPPO was lower than that of MPPO. The excited-state dipole moment of DPPO, with symmetric geometry, was calculated to be 4.1 Debye, whereas MPPO, with asymmetric geometry, had a value of 7.0 Debye. The charge-carrier mobility of both compounds was similar. We fabricated non-doped organic light-emitting diodes (OLEDs) using D-A type molecules as an emitting layer. The current efficiency of the DPPO-based device was 7.8 cd/A, and the external quantum efficiency was 2.4% at 100 cd/m2, demonstrating significantly improved performance compared to the MPPO-based device. The photophysical and electroluminescence (EL) characteristics of the two D-A type molecules showed that molecular symmetry, as well as the lowered LUMO level of DPPO, played critical roles in the enhancement of EL performance.