• Title/Summary/Keyword: Molecular Characteristics

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Molecular and Biochemical Characterization of a Novel Intracellular Low-Temperature-Active Xylanase

  • Zhou, Junpei;Dong, Yanyan;Tang, Xianghua;Li, Junjun;Xu, Bo;Wu, Qian;Gao, Yajie;Pan, Lu;Huang, Zunxi
    • Journal of Microbiology and Biotechnology
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    • v.22 no.4
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    • pp.501-509
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    • 2012
  • A 990 bp full-length gene (xynAHJ2) encoding a 329-residue polypeptide (XynAHJ2) with a calculated mass of 38.4 kDa was cloned from Bacillus sp. HJ2 harbored in a saline soil. XynAHJ2 showed no signal peptide, distinct amino acid stretches of glycoside hydrolase (GH) family 10 intracellular endoxylanases, and the highest amino acid sequence identity of 65.3% with the identified GH 10 intracellular mesophilic endoxylanase iM-KRICT PX1-Ps from Paenibacillus sp. HPL-001 (ACJ06666). The recombinant enzyme (rXynAHJ2) was expressed in Escherichia coli and displayed the typical characteristics of low-temperature-active enzyme (exhibiting optimum activity at $35^{\circ}C$, 62% at $20^{\circ}C$, and 38% at $10^{\circ}C$; thermolability at ${\geq}45^{\circ}C$). Compared with the reported GH 10 low-temperature-active endoxylanases, which are all extracellular, rXynAHJ2 showed low amino acid sequence identities (<45%), low homology (different phylogenetic cluster), and difference of structure (decreased amount of total accessible surface area and exposed nonpolar accessible surface area). Compared with the reported GH 10 intracellular endoxylanases, which are all mesophilic and thermophilic, rXynAHJ2 has decreased numbers of arginine residues and salt bridges, and showed resistance to $Ni^{2+}$, $Ca^{2+}$, or EDTA at 10 mM final concentration. The above mechanism of structural adaptation for low-temperature activity of intracellular endoxylanase rXynAHJ2 is different from that of GH 10 extracellular low-temperature-active endoxylanases. This is the first report of the molecular and biochemical characterizations of a novel intracellular low-temperature-active xylanase.

Desorption of organic Compounds from the Simulated Soils by Soil Vapor Extraction (인공토양으로부터 토양증기추출법에 의한 유기화합물의 탈착 현상에 관한 실험 연구)

  • 이병환;이종협
    • Journal of Korea Soil Environment Society
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    • v.3 no.2
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    • pp.101-114
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    • 1998
  • Soil vapor extraction (SVE) is known to be an effective process to remove the contaminants from the soils by enhancing the vaporization of organic compounds using forced vapor flows or applying vacuum through soils. Experiments are carried out to investigate the effects of the organic contaminants, types of soils, and water contents on the removal efficiency with operating time. In the study, simulated soils include the glass bead which has no micropore, sand and molecular sieve which has a large volume of micropores. As model organic pollutants, toluene, methyl ethyl ketone, and trichloroethylene are selected. Desorption experiments are conducted by flowing nitrogen gas. Under the experimental conditions, it is found that there are linear relationships between logarithm of removal efficiency and logarithm of number of pore volumes. The number of pore volumes are defined as the total amount of air flow through the soil column divided by the pore volume of soil column. For three organic compounds studied, the removal rate is slow for no water content, while the number of pore volumes for removal of organic compounds are notably reduced for water contents up to 37%. For the removal of dense organic compound, such as trichloroethylene, a large number of pore volumes are needed. Also, the effects of the characteristics of simulated soils on the removal efficiency of organic compounds are studied. After the characterization of soil surface, porosity of soil columns and types of contaminants, the results could provide a basis for the design of SVE process.

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Phenotypic and Molecular Characteristics of Children with Progressive Familial Intrahepatic Cholestasis in South China

  • Zhang, Wen;Lin, Ruizhu;Lu, Zhikun;Sheng, Huiying;Xu, Yi;Li, Xiuzhen;Cheng, Jing;Cai, Yanna;Mao, Xiaojian;Liu, Li
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.23 no.6
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    • pp.558-566
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    • 2020
  • Purpose: Progressive familial intrahepatic cholestasis (PFIC) is a rare genetic autosomal recessive disease caused by mutations in ATP8B1, ABCB11 or ABCB4. Mutational analysis of these genes is a reliable approach to identify the disorder. Methods: We collected and analyzed relevant data related to clinical diagnosis, biological investigation, and molecular determination in nine children carrying these gene mutations, who were from unrelated families in South China. Results: Of the nine patients (five males, four females) with PFIC, one case of PFIC1, four cases of PFIC2, and four cases of PFIC3 were diagnosed. Except in patient no. 8, jaundice and severe pruritus were the major clinical signs in all forms. γ-glutamyl transpeptidase was low in patients with PFIC1/PFIC2, and remained mildly elevated in patients with PFIC3. We identified 15 different mutations, including nine novel mutations (p.R470HfsX8, p.Q794X and p.I1170T of ABCB11 gene mutations, p.G319R, p.A1047P, p.G1074R, p.T830NfsX11, p.A1047PfsX8 and p.N1048TfsX of ABCB4 gene mutations) and six known mutations (p.G446R and p.F529del of ATP8B1 gene mutations, p.A588V, p.G1004D and p.R1057X of ABCB11 gene mutations, p.P479L of ABCB4 gene mutations). The results showed that compared with other regions, these three types of PFIC genes had different mutational spectrum in China. Conclusion: The study expands the genotypic spectrum of PFIC. We identified nine novel mutations of PFIC and our findings could help in the diagnosis and treatment of this disease.

Growth of Hexagonal Boron Nitride Thin Films on Silicon Using a Single Source Precursors

  • Boo, Jin-Hyo;Lee, Soon-Bo;Casten Rohr;Wilson Ho
    • Proceedings of the Korean Vacuum Society Conference
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    • 1998.02a
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    • pp.120-120
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    • 1998
  • Boron nitride (BN) films have attracted a growing interest for a variety of t technological applications due to their excellent characteristics, namely hardness, c chemical inertness, and dielectrical behavior, etc. There are two crystalline phases 1551; of BN that are analogous to phases of carbon. Hexagonal boron nitride (h-BN) has a a layered s$\sigma$ucture which is spz-bonded structure similar to that of graphite, and is t the stable ordered phase at ambient conditions. Cubic boron nitride (c-BN) has a z zinc blende structure with sp3-bonding like as diamond, 따ld is the metastable phase a at ambient conditions. Among of their prototypes, especially 삼Ie c-BN is an i interesting material because it has almost the same hardness and thermal c conductivity as di없nond. C Conventionally, significant progress has been made in the experimental t techniques for synthesizing BN films using various of the physical vapor deposition 밍ld chemical vapor deposition. But, the major disadvantage of c-BN films is that t they are much more difficult to synthesize than h-BN films due to its narrow s stability phase region, high compression stress, and problem of nitrogen source c control. Recent studies of the metalorganic chemical vapor deposition (MOCVD) of I III - V compound have established that a molecular level understanding of the d deposition process is mandatory in controlling the selectivity parameters. This led t to the concept of using a single source organometallic precursor, having the c constituent elements in stoichiometric ratio, for MOCVD growth of 삼Ie required b binary compound. I In this study, therefore, we have been carried out the growth of h-BN thin f films on silicon substrates using a single source precursors. Polycrystalline h-BN t thin films were deposited on silicon in the temperature range of $\alpha$)() - 900 $^{\circ}$C from t the organometallic precursors of Boron-Triethylamine complex, (CZHs)3N:BRJ, and T Tris(dimethylamino)Borane, [CH3}zNhB, by supersonic molecular jet and remote p plasma assisted MOCVD. Hydrogen was used as carrier gas, and additional nitrogen w was supplied by either aDlIDonia through a nozzle, or nitrogen via a remote plasma. T The as-grown films were characterized by Fourier transform infrared spectroscopy, x x-ray pthotoelectron spectroscopy, Auger electron spectroscopy, x-ray diffraction, t transmission electron diffraction, optical transmission, and atomic force microscopy.roscopy.

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Synthesis and Properties of Copolyester Resin for Precoated MeTAL (Precoated Metal용 Copolyester Resin의 합성 및 물성)

  • Park, Lee Soon;Lee, Tae Hyung;Kim, Soon Hak;Chang, Jin Gyu
    • Applied Chemistry for Engineering
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    • v.10 no.3
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    • pp.427-431
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    • 1999
  • Copolyester resins for the coil coating process of aluminium and steel strip were synthesized and their thermal properties, molecular weight and solvent solution characteristics were examined. Copolyesters were obtained by two step reactions. The first step was to prepare bishydroxyethyl terephthalate (BHET), bishydroxyneopentyl terephthalate (BHNPT), bishydroxyethyl isophthalate (BHEI), bishydroxyneopentyl sebacate (BHNPS), bishydroxyneopentyl adipicate (BHNPA) and bishydroxyethyl adipicate (BHEA) oligomers by esterification reactions. The second step was the polycondensation reaction utilizing those oligomers to obtain relatively high molecular weight copolyesters (Mw = 30,000~59,000 g/mol) as measured by GPC. These copolyesters were amorphous polymers as shown by DSC without $T_m$ peaks probably due to the kink structure introduced by BHET oligomer and relatively large free volume by bulky BHNPT and BHNPS oligomers. The copolyester samples with half of BHET oligomer substituted by BHNPT while keeping BHEI (0.3 mole) and BHNPS (0.1 mole) ratio constant showed glass transition temperature above $40^{\circ}C$ and good solubility in toluene both at low ($-5^{\circ}C$) temperature and room temperature.

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The Study of Genetic Diversity and Population Structure of the Korean Fleshy Shrimp, Fenneropenaeus chinensis, Using Newly Developed Microsatellite Markers (새로 개발한 미세위성체 마커를 이용한 한국 대하의 유전다양성 및 집단구조)

  • Shin, Eun-Ha;Kong, Hee Jeong;Nam, Bo-Hye;Kim, Young-Ok;Kim, Bong-Seok;Kim, Dong-Gyun;An, Cheul Min;Jung, Hyungtaek;Kim, Woo-Jin
    • Journal of Life Science
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    • v.25 no.12
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    • pp.1347-1353
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    • 2015
  • The fleshy shrimp, Fenneropenaeus chinensis, is the family of Penaeidae and one of the most economically important marine culture species in Korea. However, its genetic characteristics have never been studied. In this study, a total of 240 wild F. chinensis individuals were collected from four locations as follows: Narodo (NRD, n = 60), Beopseongpo (BSP, n = 60), Chaesukpo (CSP, n = 60), and Cheonsuman (CSM, n = 60). Genetic variability and the relationships among four wild F. chinensis populations were analyzed using 13 newly developed microsatellite loci. Relatively high levels of genetic variability (mean allelic richness = 16.87; mean heterozygosity = 0.845) were found among localities. Among the 52 population loci, 13 showed significant deviation from the Hardy–Weinberg equilibrium. Neighbor-joining, principal coordinate, and molecular variance analyses revealed the presence of three subpopulations (NRD, CSM, BSP and CSP), which was consistent with clustering based on genetic distance. The mean observed heterozygosity values of the NRD, CSM, BSP, and CSP populations were 0.724, 0.821, 0.814, and 0.785 over all loci, respectively. These genetic variability and differentiation results of the four wild populations can be applied for future genetic improvement using selective breeding and to design suitable management guidelines for Korean F. chinensis culture.

Analysis of the Correlation between Expressions of HSP90α, HSP90β, and GRP94, and the Clinicopathologic Characteristics in Tissues of Non-Small Cell Lung Cancer Patients (비소세포 폐암 환자 조직에서 Hsp90α, Hsp90β, GRP94의 발현과 임상병리학적 특성과의 상관관계 분석)

  • Kim, Mi Kyeong
    • Korean Journal of Clinical Laboratory Science
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    • v.49 no.4
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    • pp.460-469
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    • 2017
  • Heat shock proteins (HSPs) are induced as a self-defense mechanism of cells when exposed to various external stresses, such as high fever, infection, free radicals, and heavy metals. They affect the prognosis in the process of tumor formation. HSP is classified into four families: HSP27, HSP60, HSP90, and HSP100, depending on molecular weight. Heat shock protein 90 (HSP90), a molecular chaperone, plays an important role in the cellular protection against various stressful stimuli and in the regulation of cell cycle progression and apoptosis. In the present study, we assessed the differential expression of HSP90 family proteins in non-small cell lung cancer (NSCLC), and the correlation of their expression levels with clinicopathologic factors and patient survival rates. The result of this study can be summarized as follows; $HSP90{\alpha}$ showed higher expression in patients with no lymphovascular invasion (p=0.014). $HSP90{\beta}$ showed a higher expression of squamous cell carcinoma (p=0.003), and an over expression of glucose-related protein (GRP94) was significantly associated with poor differentiation (p=0.048). However, none of the HSP90 proteins showed a significant association with the survival status in patients with NSCLC. This study also indicates that $HSP90{\alpha}$ might contribute more to the carcinogenesis of NSCLC than $HSP90{\beta}$, and GRP94 and isoform selectivity should be considered when HSP90 inhibitors are studied or utilized in the treatment of NSCLC.

Immunological Characterization of Antigens from Custiceycus and Sparganum and Their Application to Immunodiagnosis 1. Immunological Characteristics of Crude Antigenic Components from Cucticercus cellulosae (낭미충(Cysticercus)과 스파르가눔(Sparganum)에서 추출한 조항원의 면역학적 특성 및 면역진단에의 응용 1. 낭미충에서 추출한 조항원 성분의 면역학적 특성)

  • ;James Yang
    • Parasites, Hosts and Diseases
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    • v.26 no.4
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    • pp.245-254
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    • 1988
  • We studied the serological reaction between various antigenic components from Cysticercus cellulosae and IgG antibodies in sera of cysticercosis, sparganosis, hydatidosis patients and normal humans by ELISA and EITB. In serological tests by ELISA, we recognized cross reaction of Cysticercus antigenic components with IgG antibodies in heterologous sera such as sparganosis and hydatidosis patients or normal humans. The crude antigenic components of Cysticercus showed lower ELISA sensitivity in homologous sera from cysticercosis patients than heterologous sera from hydatidosis patients. A total of 31 polypeptide bands with 260 KDa~22 KDa molecular weights were detected by SDS-PAGE, and 11 of them showed strong intensity. Total 22 components of them were recognized by IgG antibodies in cysticercosis patients sera. However, 12 of them were recognized also by normal human sera, 11 were by sparganosis sera, and-21 were by hydatidosis patients sera. The crude antigenic components of 104 KDa, 82 KDa, 72 KDa, 59 KDa and 34 KDa molecular weights were nonspecific ones, which cross-reacted with sera of either cysticerco, =is, sparganosis, hydatidosis patients or normal humans.

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Physiological and Molecular Characterization of NAD(P)H-Nitroreductase from Stenotrophomonas sp. OK-5 (Stenotrophomonas sp. OK-5에서 분리한 NAD(P)H-Nitroreductase의 생리학적 및 분자생학적 특성 연구)

  • Ho Eun-Mi;Kahng Hyung-Yeel;Oh Kye-Heon
    • Korean Journal of Microbiology
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    • v.40 no.3
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    • pp.183-188
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    • 2004
  • Stenotrophomonas sp. OK-5 capable of degrading TNT has been found to have three nitroreductase fractions designated as NTR fractions I, II, and III. NTR in a previous study. This study was attempted to reveal physiological and molecular characteristics of NTR fractions I, II, and III in strain OK-5. Several chemicals (e.g., EDTA, NaCl, dithiothreitol, $\beta$-mercaptoethanol) were tested for their effect on enzyme activity of NTRs, demonstrating that enzyme activities of NTR fractions I, II, and III from OK-5 were inhibited in the presence of $\beta$-mercaptoethanol. Substrate specificity test showed that NTR fractions I, II, and III all have over 70% enzyme activities for nitrobenzene or RDX as a substrate. N-terminal amino acid sequence of NTR fraction I from Stenotrophomonas sp. OK-5 was $^1MSDLLNADAVVQLFRTARDS^20$ and exhibited 70% sequence homology with that of NTR from Xanthomonas campestris. NTR I gene from Stenotrophomonas sp. OK-5 (SmOK5nrI) shared extensive sequence homology in deduced amino acid sequence of PCR product with NTRs from Xanthomonas campestris (81 %), X. axonopodis (75%), Streptomyces avermitilis(30%), whereas they had low homology with that from P. putida KT2440 (pnrB) (16%).

Identification of a Causal Pathogen of Watermelon Powdery Mildew in Korea and Development of a Genetic Linkage Marker for Resistance in Watermelon (Citrullus lanatus)

  • Han, Bal-Kum;Rhee, Sun-Ju;Jang, Yoon Jeong;Sim, Tae Yong;Kim, Yong-Jae;Park, Tae-Sung;Lee, Gung Pyo
    • Horticultural Science & Technology
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    • v.34 no.6
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    • pp.912-923
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    • 2016
  • Watermelon production is often limited by powdery mildew in areas with a large daily temperature range. Development of resistant watermelon cultivars can protect against powdery mildew; however, little is known about the characteristics of its causal agents. Here, we identified the genus and race of a causal pathogen of powdery mildew in Ansung province of South Korea, and developed molecular markers for the generation of resistant watermelon cultivars. The causal pathogen was determined to be Podosphaera xanthii based on multiple sequence alignments of internal transcribed spacers (ITS) of rDNA. The physiological race was identified as 1W, and the Ansung isolate was named P. xanthii 1W-AN. Following inoculation with the identified P. xanthii 1W-AN, we found inheritance of the resistant gene fitting a single dominant Mendelian model in a segregated population ('SBA' ${\times}$ PI 254744). To develop molecular markers linked to fungus-resistant loci, random amplified polymorphic DNA (RAPD) was accomplished between DNA pooled from eight near-isogenic lines (NILs; $BC_4F_6$), originated from PI 254744 and susceptible 'SBB' watermelon. After sequencing bands from RAPD were identified in all eight NILs and PI254744, 42 sequence-characterized amplifiedregion (SCAR) markers were developed. Overall, 107 $F_2$ plants derived from $BC_4F_6$ NIL-1 ${\times}$ 'SBB' were tested, and one SCAR marker was selected. Sequence comparison between the SCAR marker and the reference watermelon genome identified three Nco I restriction enzyme sites harboring a single nucleotide polymorphism, and codominant cleavage-amplified polymorphic site markers were subsequently developed. A CAPS marker was converted to a high-resolution melt (HRM) marker, which can discriminate C/T SNP (254PMR-HRM3). The 254PMR-HRM3 marker was evaluated in 138 $F_{2:3}$ plants of a segregating population ('SBA' ${\times}$ PI254744) and was presumed to be 4.3 cM from the resistance locus. These results could ensure P. xanthii 1W-AN resistance in watermelon germplasm and aid watermelon cultivar development in marker-assist breeding programs.