• 제목/요약/키워드: Molecular Characterization

검색결과 3,085건 처리시간 0.031초

Multiparameter Flow Cytometry: Advances in High Resolution Analysis

  • O'Donnell, Erika A.;Ernst, David N.;Hingorani, Ravi
    • IMMUNE NETWORK
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    • 제13권2호
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    • pp.43-54
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    • 2013
  • Over the past 40 years, flow cytometry has emerged as a leading, application-rich technology that supports high-resolution characterization of individual cells which function in complex cellular networks such as the immune system. This brief overview highlights advances in multiparameter flow cytometric technologies and reagent applications for characterization and functional analysis of cells modulating the immune network. These advances significantly support highthroughput and high-content analyses and enable an integrated understanding of the cellular and molecular interactions that underlie complex biological systems.

Mycoflora and Enzymatic Characterization of Fungal Isolates in Commercial Meju, Starter for a Korean Traditional Fermented Soybean Product

  • Baek, Jin-Ho;So, Kum-Kang;Ko, Yo-Han;Kim, Jung-Mi;Kim, Dae-Hyuk
    • Mycobiology
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    • 제42권3호
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    • pp.291-295
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    • 2014
  • Mycoflora was assessed in the commercial meju from four well-separated geographic origins. A total of 112 fungal isolates were identified by phenotypic characteristics and molecular taxonomy using sequencing the internal transcribed spacer of the rDNA and revealed 19 species from 13 genera. Enzymatic characteristics of protease and amylase, and mycotoxin production were analyzed.

Purification and Characterization of $\beta$-Xylosidase from Trichoderma sp. SY

  • Kim, Bong-Gyu;Jung, Bo-Ra;Jung, Jae-Gyu;Hur, Hor-Gil;Ahn, Joong-Hoon
    • Journal of Microbiology and Biotechnology
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    • 제14권3호
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    • pp.643-645
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    • 2004
  • A $\beta$-xylosidase was purified from the culture of Trichoderma sp. SY. The ten-day-old culture filtrate was concentrated, followed by ion-exchange chromatography and gel filtration chromatography. As a result, $\beta$-xylosidase was finally purified about 53-fold and appeared as a single band by SDS-PAGE. The optimum pH and temperature were 5.0 and $55^{\circ}C$, respectively, and the molecular weight about 80 kDa. The purified $\beta$-xylosidase was found to be inhibited by various metal ions but not inhibited by xylose.

Synthesis and Characterization of Star-shaped Aliphatic Polyester

  • Shin, Young-Cheol;Park, Kil-Yeong;Jin, Moon-Young;Hong, Sung-Kwon;Donghyun Cho;Taihyun Chang;Moonhor Ree
    • Macromolecular Research
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    • 제9권2호
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    • pp.100-106
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    • 2001
  • Several star-polymers with aliphatic cores were synthesized by ring opening polymerization (ROP) of $\xi$-caprolactone using stannous 2-ethyl-hexanoate as a catalyst. The star-polymers were thoroughly analyzed by MALDI-TOF mass spectrometry, temperature gradient interaction chromatography and $\^$13/C-NMR to obtain detailed information of the molecular structure. The imperfection of the star-polymers seemed to be originated from restricted participation of sterically hindered hydroxyl groups of initiator. The synthesized star-polymers had narrow molecular weight distributions. Various reaction conditions to control the imperfection were studied.

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Purification and Characterization of Glyoxalase I from Chlamydomonas reinhardtii

  • Hwang, Sun-Jun;Chai, Young-Gyu
    • BMB Reports
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    • 제29권4호
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    • pp.294-299
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    • 1996
  • Glyoxalase I (Ee 4.4.1.5, lactoylglutathione lyase) from Chlamydomonas reinhardtii was purified to homogeneity by ammonium sulfate fractionation, anion-exchange chromatography, hydrophobic interaction chromatography, and affinity chromatography on S-hexylglutathione agarose. The purified enzyme was judged to be homogeneous on SDS-PAGE, and consisted of a single polypeptide chain with a relative molecular weight of 24,000. The enzyme was most active at $40^{\circ}C$ and pH 7.5. It was catalytically most active with methylglyoxal as substrate. A number of properties of the Chlamydomonas glyoxalase I enzyme, such as substrate specificity, molecular mass, kinetic parameters, pi, metal ion effect, have been determined and compared with those reported for preparations from other sources. It had somewhat different characteristics from mammalian enzymes.

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Structural Characterization of Branched Polyesters Using TOF-SIMS Combined with Transesterification

  • Lee, Yeonhee;Seunghee Han;Yoon, Jung-Hyeon;Hyuneui Lim;Moojin Suh
    • 한국진공학회:학술대회논문집
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    • 한국진공학회 1999년도 제17회 학술발표회 논문개요집
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    • pp.203-203
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    • 1999
  • Mass spectrometry technique provides the molecular weight distribution, data on the sequence of repeat units, polymer additives, and impurities, and structural information. time-of-Flight secondary Ion Mass Spectrometry (TOF-SIMS) has been used for structural characterization of various polymers1-2. the masses of repeat units and terminal groups and molecular weight distributions of polymers have been determined from their TOF-SIMS spectra. TOF-SMIS provides good sensitivity and structural specificity for high mass ions so that intact oligomers and large polymer fragments are observed. In this study, we investigated the detailed structural information on the oligomers and fragment ions of branched poly(1,3-butylene adipate) and branched poly[di(ethylene glycol) adipate] and the transesterification products of branched polyesters with trifluoroacetic acid or chloro difluoroacetic acid. Branched polyesters were chosen because they are important polymers but difficult to characterize; thus branched polyesters provide challanging test for TOF-SIMS. TOF-SIMS spectra of polyesters are obtained from thin polymer films cast from solution on a silver substrate. A good solvent for a polumer solution disrupts intermolecular forces between polymer chains but leaves the polumer intact. Transesterification reactions are potentially useful for characterization of high molecular weight and intractable polyesters. Transesterification products of polyesters and trifluoroacetic acid or an integral number of polyester repeat units and an additional diol. The progress of such reactions was monitored using peak intensities of reactants and products in TOF-SIMS spectra. The increasing abundance of tagged ions indicates that the reaction has progressed with time.

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Molecular characterization of lepidopteran pest-resistant transgenic rice events expressing synthetic Cry1Ac

  • Lee, Kyeong-Ryeol;Shin, Kong Sik;Suh, Seok Cheol;Kim, Ki Young;Jeon, Yong Hee;Park, Beom Seok;Kim, Ju-Kon;Kweon, Soon-Jong;Lee, Yeon-Hee
    • Plant Biotechnology Reports
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    • 제3권4호
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    • pp.317-324
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    • 2009
  • The insecticidal toxin gene of Bacillus thuringiensis (Bt) is one of the most commonly used in the development of genetically modified (GM) crops. In this research, we analyzed Bt rice showing lepidopteran pest-resistance. The Bt gene is a synthetic Cry1Ac composed of optimal codons for plants, and the Bt protein is targeted to the chloroplast by a transit peptide. Three Cry1Ac rice events (C103-3, C127-1, and C7-1) were analyzed for molecular characterization. C103-3 contains two copies of T-DNA where the left border (LB) region is truncated. Both C7-1 and C127-1 have a single copy of T-DNA, but a part of the vector backbone DNA is inserted into the genome of C127-1; thus, only C7-1 had intact T-DNA. Progenies of C7-1 crossed with the original cultivar, Nakdong, and double-haploid lines from anther culture of lines crossed with the elite cultivar, Dongjin, were analyzed for T-DNA flanking genomic DNA and genotyping. Results showed that an intact T-DNA region without the vector backbone was inserted into the genome and was stably inherited through generations. The C7-1 homozygous event could be used as breeding material to develop GM rice with pest resistance.

Characterization of Alpha Amylase Producing Thielaviopsis ethacetica and Its Raw Starch Hydrolyzing Ability on Different Agricultural Substrates

  • Dissanayaka, Dissanayaka M.S.;De Silva, Sembukuttige N.T.;Attanayaka, D.P.S.T.G.;Kurera, Mihidukulasuriya J.M.S.;Fernando, Charakrawarthige A.N.
    • 한국미생물·생명공학회지
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    • 제47권3호
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    • pp.412-422
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    • 2019
  • The present study reports the morphological and molecular characterization of the fungal strain, CMSS06 and evaluates its raw starch hydrolyzing ability in four different agricultural substrates (rice bran, banana peel, cassava tubers, and coconut water). The potential use of each agricultural substrate to replace the expensive fermentation media was evaluated with six different fermentation media: rice bran (RB), banana peel (BP), cassava starch (CS), cassava in coconut water (CSCW), cassava in modified coconut water (CMCW), and pure Coconut water (CW). The fungal strain CMSS06 was identified as Thielaviopsis ethacetica by the analysis of the ITS sequences. The T. ethacetica alpha amylase enzyme exhibited maximum alpha amylase activity at 72 h, pH 7.0, and $40^{\circ}C$ on soluble starch. This species resulted in the highest enzyme activity (mU/ml) of 26.06, 10.89, 58.82, 14.2, and 54.67 with the RB, BP, CS, CSCW, and CMCW fermentation media, respectively. The results indicate that CS can be used as a carbon substrate and CMCW can be used to accelerate the fermentation by T. ethacetica. The enzyme was partially purified by 40-60% ammonium sulphate fraction, and it showed total enzyme activity, total protein content, specific activity, purification fold, and a recovery of 2400 mU, 30 mg, 80 mU/mg, 2.7, and 71.1%, respectively. The molecular mass of the T. ethacetica alpha amylase was estimated on SDS-PAGE, and two bands around 50 kDa and 70 kDa were identified. The present study implies that T. ethacetica can produce alpha amylase, and it can be used to hydrolyze raw starch during the fermentation processes.

Purification and Physicochemical Characterization of a Recombinant Phospholipid Hydroperoxide Glutathione Peroxidase from Oryza sativa

  • Wang, Zebin;Wang, Feng;Duan, Rui;Liu, Jin-Yuan
    • BMB Reports
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    • 제40권3호
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    • pp.412-418
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    • 2007
  • Phospholipid hydroperoxide glutathione peroxidase (PHGPx) is an unique antioxidant enzyme that directly reduces lipid hydroperoxides in biomembranes. In the present work, the entire encoding region for Oryza sativa PHGPx was expressed in Escherichia coli M15, and the purified fusion protein showed a single band with 21.0 kD and pI = 8.5 on SDS- and IFE-PAGE, respectively. Judging from CD and fluorescence spectroscopy, this protein is considered to have a well-ordered structure with 12.2% $\alpha$-helix, 30.7%$\beta$-sheet, 18.5% $\delta$-turn, and 38.5% random coil. The optimum pH and temperature of the enzyme activity were pH 9.3 and 27$^{\circ}C$. The enzyme exhibited the highest affinity and catalytical efficiency to phospholipid hydroperoxide employing GSH or Trx as electron donor. Moreover, the protein displayed higher GSH-dependent activity towards t-Butyl-OOH and $H_2O_2$. These results show that OsPHGPx is an enzyme with broad specificity for hydroperoxide substrates and yielded significant insight into the physicochemical properties and the dynamics of OsPHGPx.