• Title/Summary/Keyword: Chain elongation

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Cloning, Purification, and Characterization of a New DNA Polymerase from a Hyperthermophilic Archaeon, Thermococcus sp. NA1

  • Kim, Yun-Jae;Lee, Hyun-Sook;Bae, Seung-Seob;Jeon, Jeong-Ho;Lim, Jae-Kyu;Cho, Yon-A;Nam, Ki-Hoon;Kang, Sung-Gyun;Kim, Sang-Jin;Kwon, Suk-Tae;Lee, Jung-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.17 no.7
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    • pp.1090-1097
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    • 2007
  • Genomic analysis of Thermococcus sp. NA1 revealed the presence of a 3,927-base-pair (bp) family B-type DNA polymerase gene, TNA1_pol. TNA1_pol, without its intein, was overexpressed in Escherichia coli, purified using metal affinity chromatography, and characterized. TNA1_pol activity was optimal at pH 7.5 and $75^{\circ}C$. TNA1_pol was highly thermostable, with a half-life of 3.5h at $100^{\circ}C$ and 12.5h at $95^{\circ}C$. Polymerase chain reaction parameters of TNA1_pol such as error-rate, processivity, and extension rate were measured in comparison with rTaq, Pfu, and KOD DNA polymerases. TNA1_pol averaged one incorrect bp every 4.45 kilobases (kb), and had a processivity of 150 nucleotides (nt) and an extension rate of 60 bases/s. Thus, TNA1_pol has a much faster elongation rate than Pfu DNA polymerase with 7-fold higher fidelity than that of rTaq.

Preparation of Sulfur Crosslinkable EVA and Blend With Rubbers (황가황형 EVA의 제조 및 고무와의 블렌드)

  • Jin, Je-Yong;Choi, Sei-Young
    • Elastomers and Composites
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    • v.34 no.3
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    • pp.229-238
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    • 1999
  • In this study, double bond, unsaturated group was introduced to the main chain of EVA by chemically treating EVA, nondiene polymer. Benzene sulfonic acid, ENB and DCPD were used as a third element. Also, from blending CR and SBR, conventional synthetic rubber we prepared vulcanizates and examined their physical properties. The datum lead to the following conclusion that some problems were modified; limited temperature in use and mechanical properties like hardness, tensile strength, tensile stress, and elongation rate of thermoplastic EVA, keeping the following advantages of original EVA; green strength, injection molding by Pressure, adhesion, tackiness, dimensional stability, and ozone resistance, etc. It is expected that continuous research of the modification between nondiene and diene polymer will improve what were shown disadvantages in synthetic polymer; processing, oxidation resistance, and adhesion. In addition, it will be possible to continue process of rubber products by utilizing possible fluidity for fusion of EVA.

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A Study on Processing and Physical Properties of Isoprene Rubber Involving Norbornene Dialkyl Ester (Norbornene Dialkyl Ester가 첨가된 Isoprene Rubber의 가공성 및 물성에 관한 연구)

  • Jeong, Hye-in;Jo, Nam-chol;Woo, Je-Wan
    • Applied Chemistry for Engineering
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    • v.27 no.3
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    • pp.259-264
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    • 2016
  • In this study, we applied six different norbornene dialkyl esters as a plasticizer to an isoprene rubber (IR) and evaluated replaceability of DEHP which is an endocrine disruptor. IR test sheets were prepared by blending IR, norbornene dialkyl ester, vulcanizing agent, etc. and processing properties of the IR were evaluated by measuring Toque, scorch time, cure time and mooney viscosity. Mechanical properties of IR test sheet including hardness, tensile strength, 100% modulus and elongation were also measured and the physical properties of norbornene dialkyl ester applied as a plasticizer were compared to those using DEHN. Both the maximum and minimum toque for the norbornene dialkyl ester as a plasticizer were similar to those of using DEHP. In addition, the scorch and cure time of the former were slightly longer than those of the latter. The mooney viscosity for the case of DEHN was slightly lower than that of the latter. DEHN was also superior to DEHP in terms of processing. The hardness and thermal properties of all IR test sheets were measured to be similar to each other. The linear alkyl chain of norbornene compounds also exhibited good tensile characteristics.

Keeping house: evaluation of housekeeping genes for real-time PCR in the red alga, Bostrychia moritziana (Florideophyceae)

  • Shim, Junbo;Shim, Eunyoung;Kim, Gwang Hoon;Han, Jong Won;Zuccarello, Giuseppe C.
    • ALGAE
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    • v.31 no.2
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    • pp.167-174
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    • 2016
  • Biological response of cells to variable conditions should affect the expression level of certain genes. Quantification of these changes in target genes needs stable internal controls. Real-time quantitative polymerase chain reaction (PCR) has traditionally used reference or ‘housekeeping’ genes, that are considered to maintain equal expression in different conditions, to evaluate changes in target genes between samples and experimental conditions. Recent studies showed that some housekeeping genes may vary considerably in certain biological samples. This has not been evaluated in red algae. In order to identify the optimal internal controls for real-time PCR, we studied the expression of eleven commonly used housekeeping genes; elongation factor 1-alpha, glyceraldehyde-3-phosphate dehydrogenase, β-actin, polyubiquitin, 30S ribosomal gene, 60S ribosomal gene, beta-tubulin, alpha-tubulin, translation initiation factor, ubiquitin-conjugating enzyme, and isocitrate dehydrogenase in different life-history stages of Bostrychia moritziana. Our results suggest that glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and 30S ribosomal gene, have the most stable gene expression levels between the different life history stages (male, female, carposporophyte, and tetrasporophyte), while the other genes are not satisfactory as internal controls. These results suggest that the combinations of GAPDH and 30S would be useful as internal controls to assess expression level changes in genes that may control different physiological processes in this organism or that may change in different life history stages. These results may also be useful in other red algal systems.

Gamma fatty acid : A review (감마지방산 : 리뷰)

  • Park, Byung-Sung
    • Journal of the Korean Applied Science and Technology
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    • v.25 no.4
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    • pp.446-458
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    • 2008
  • Essential fatty acids (EFA) are fatty acids that must be obtained from the diet because they can not be biosynthesized by human or animals. Gamma fatty acids contain gamma-linolenic acid (GLA, 18:3n-6) and dihomo-gamma-linolenic acid (DHGLA, 20:3n-6) as intermediate metabolites of linoleic acid (LA, 18:2n-6), which is an EFA found in vegetable oils. GLA is an important essential fatty acid that is required by human and animals to function normally. Recently, studies have indicated that GLA may be an essential component of the cell membrane, as well as an active component of dietary supplements and medicine. GLA must beadministered through the diet because it is converted into DHGLA in the body quickly and completely. DHGLA is a key material involved in the metabolism of LA. GLA is biosysthesized by the rate limiting step of ${\Deltac}^6$-desaturase, which is an enzyme that desaturates LA, there by allowing it to be converted into DHGLA via chain elongation. In addition, DHGLA exerts bioactive effects via action as a precursor of eicosanoid series 1. Breast milk contains an abundant amount of GLA; however, GLA is also available directly in evening primrose oil, black currant seed oil, borage oil and hemp seed oil. In addition, GLA enriched animal and plant can be produced using biotechnology, and highly pure GLA can be extracted using supercritical fluids, such as supercritical carbon dioxide, which will allow economically feasible production of GLA for use in medicines.

Structure Development in Drawn Poly(trimethylene terephthalate) (연신에 의한 폴리(트리메틸렌 테레프탈레이트)의 구조 변형)

  • 전병환;김환기;강호종
    • Polymer(Korea)
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    • v.27 no.5
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    • pp.477-483
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    • 2003
  • The structure development of drawn poly(trimethylene terephthalate) PTT as a function of draw down ratio and drawing temperature was studied. The special effort was made to find out the effect of structural development on thermal properties and crystallinity in drawn PTT. The changes in shrinkage ratio and mechanical properties were understood base on the level of crystallinity and orientation of the drawn PTT. The stress induced crystallization caused the increase in glass transition temperature and the decrease in cold crystallization temperature and enthalpy. The crystallinity and orientation were dependent upon the level of applied stress level as well as chain flexibility at high drawing temperature. The drawing resulted in the increase of shrinkage ratio but it was minimized by increasing of crystallinity. The development of orientation resulted in increasing modulus and tensile strength while decreasing elongation at break.

Kinematics of filament stretching in dilute and concentrated polymer solutions

  • McKinley, Gareth H.;Brauner, Octavia;Yao, Minwu
    • Korea-Australia Rheology Journal
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    • v.13 no.1
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    • pp.29-35
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    • 2001
  • The development of filament stretching extensional rheometers over the past decade has enabled the systematic measurement of the transient extensional stress growth in dilute and semi-dilute polymer solutions. The strain-hardening in the extensional viscosity of dilute solutions overwhelms the perturbative effects of capillarity, inertia & gravity and the kinematics of the extensional deformation become increasingly homogeneous at large strains. This permits the development of a robust open-loop control algorithm for rapidly realizing a deformation with constant stretch history that is desired for extensional rheometry. For entangled fluids such as concentrated solutions and melts the situation is less well defined since the material functions are governed by the molecular weight between entanglements, and the fluids therefore show much less pronounced strain-hardening in transient elongation. We use experiments with semi-dilute/entangled and concentrated/entangled monodisperse polystyrene solutions coupled with time-dependent numerical computations using nonlinear viscoelastic constitutive equations such as the Giesekus model in order to show that an open-loop control strategy is still viable for such fluids. Multiple iterations using a successive substitution may be necessary, however, in order to obtain the true transient extensional viscosity material function. At large strains and high extension rates the extension of fluid filaments in both dilute and concentrated polymer solutions is limited by the onset of purely elastic instabilities which result in necking or peeling of the elongating column. The mode of instability is demonstrated to be a sensitive function of the magnitude of the strain-hardening in the fluid sample. In entangled solutions of linear polymers the observed transition from necking instability to peeling instability observed at high strain rates (of order of the reciprocal of the Rouse time for the fluid) is directly connected to the cross-over from a reptative mechanism of tube orientation to one of chain extension.

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An Efficient Plant Regeneration and Transformation System of Robinia pseudoacacia var. umbraculifera for Phytoremediation

  • Kwon, Hye-Jin;Woo, Seong-Min;Seul, Eun-Jun;Kim, Teh-Ryung;Shin, Dong-Un;Kim, Hag-Hyun
    • Journal of Plant Biotechnology
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    • v.34 no.4
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    • pp.293-298
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    • 2007
  • Robinia pseudoacacia var. umbraculifera, commonly called umbrella black locust were regenerated after co-cultivation of internode segments with Agrobacterium tumefaciens which included yeast cadmium factor 1 (YCF 1) gene. The tolerance to cadmium and lead for plants can be increased by the YCF1 gene expression. Moreover, the recent studies have shown that YCF1 gene transgenic plants increase the accumulation of cadmium and lead into plant vacuoles. The effect of plant growth regulator such as 2,4-dichlorophenoxyacetic acid (2,4-D), ${\alpha}$-naphthaleneacetic acid (NAA), 6-benzyladenine (BA), and thidiazuron (TDZ) were studied to evaluate the propagation of plants through internode explants. The efficient induction of multiple adventitious shoots and callus were observed on a medium supplemented with 0.1 mg/L TDZ + 0.2 mg/L BA. To induce shoot elongation and rooting, regenerated shoots were transferred into basal MS medium without any plant growth regulator. Successful Agrobacterium tumefaciens mediated transformation was obtained by 20 min vacuum-infiltration with $50{\mu}M$ acetosyringone on the optimal multiple shoot induction medium with 30 mg/L hygromycin and 300 mg/L cefotaxime. To confirm the integration and expression of transgene, Polymerase Chain Reaction (PCR) and Reverse Transcriptase PCR (RT-PCR) were performed with specific primers. The frequency of transformation was approximately 18.94%. This study can be used to genetic engineering of phytoremediator.

Genome wide association study of fatty acid composition in Duroc swine

  • Viterbo, Vanessa S.;Lopez, Bryan Irvine M.;Kang, Hyunsung;Kim, Hoonseop;Song, Choul-won;Seo, Kang Seok
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.8
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    • pp.1127-1133
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    • 2018
  • Objective: Genome wide association study was conducted to identify and validate candidate genes associated with fatty acid composition of pork. Methods: A total of 480 purebreed Duroc pigs were genotyped using IlluminaPorcine60k bead chips while the association test was implemented following genome-wide rapid association using Mixed Model and Regression-Genomic Control (GRAMMAR-GC) approach. Results: A total of 25, 29, and 16 single nucleotide polymorphisms (SNPs) were significantly associated with stearic (18:0), oleic (18:1) and saturated fatty acids (SFA), respectively. Genome wide significant variants were located on the same region of swine chromosome 14 (SSC14) that spanned from 120 to 124 Mb. Top SNP ALGA008191 was located at 5 kb near the stearoyl-CoA desaturase (SCD) gene. This gene is directly involved in desaturation of stearic acid into oleic acid. General relationship of significant SNPs showed high linkage disequilibrium thus genome-wide signals was attributed to SCD gene. However, understanding the role of other genes like elongation of very long chain fatty acids-3 (ELOVL3) located on this chromosomal segment might help in further understanding of metabolism and biosynthesis of fatty acids. Conclusion: Overall, this study provides evidence that validates SCD gene as strong candidate gene associated with fatty acid composition in Duroc pigs. Moreover, this study confirms significant SNPs near ELOVL3 gene.

Stretchable Deformation-Resistance Characteristics of the Stiffness-Gradient Stretchable Electronic Packages Based on PDMS (PDMS 기반 강성도 경사형 신축 전자패키지의 신축변형-저항 특성)

  • Park, Dae Ung;Oh, Tae Sung
    • Journal of the Microelectronics and Packaging Society
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    • v.26 no.4
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    • pp.47-53
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    • 2019
  • Stiffness-gradient stretchable electronic packages of the soft PDMS/hard PDMS/PTFE structure were processed using the polydimethylsiloxane (PDMS) as the base substrate and the more stiff polytetrafluoroethylene (PTFE) as the island substrate, and their stretchable deformation-resistance characteristics were characterized. The flip-chip joints, formed by bonding the chip bumps of 50 ㎛-diameter onto the PDMS/PTFE substrate pads, exhibited an average contact resistance of 96 mΩ. When the stretchable package of the soft PDMS/hard PDMS/PTFE structure was deformed to 30% elongation, the strain on the PTFE was restrained to 1%, resulting in a negligible resistance increase of 1% in the daisy-chain circuit formed on the PTFE island substrate. The circuit resistance increased for 1.7% after 2,500 cycles of 0~30% stretchable deformation.