• Title/Summary/Keyword: Amber

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Establishment of an In Vivo Report System for the Evaluation of Amber Suppression Activity in Escherichia coli (대장균에서 비천연 아미노산의 위치특이적 삽입을 위한 Amber Suppressor tRNA와 Aminoacyl-tRNA Synthetase의 Amber Suppression 활성측정시스템 개발)

  • Kim, Kyung-Tae;Park, Mi-Young;Park, Jung-Chan
    • Korean Journal of Microbiology
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    • v.45 no.2
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    • pp.215-221
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    • 2009
  • Site-specific incorporation of unnatural amino acids into proteins in vivo can be achieved by co-expression of an orthogonal pair of suppressor tRNA and engineered aminoacyl-tRNA synthetase (ARS) that specifically ligates an unnatural amino acid to the suppressor tRNA. As a step to establish this technique, here we generated an Escherichia coli reporter strain DH10B(Tn:lacZam) by integrating amber mutated lacZ gene into the chromosome of E. coli DH10B strain. In vivo expression of E. coli amber suppressor $tRNA^{Gln}$ produced blue colonies in culture plates containing X-Gal as well as dramatically increased $\beta$-galactosidase activity. In addition, expression of an orthogonal pair of Saccharomyces cerevisiae suppressor $tRNA^{Tyr}$ and tyrosyl-tRNA synthetase also produced blue colonies as well as moderate increase of $\beta$-galactosidase activity. These data demonstrate that our reporter strain will provide an efficient method to assess amber suppression in both qualitative and quantitative manners.

Analysis of changes in composition of amber with ageing using pyrolysis/GC/MS (열분해/GC/MS를 이용한 열화 호박(amber)의 성분 변화 분석)

  • Park, Jongseo
    • Analytical Science and Technology
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    • v.26 no.3
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    • pp.190-198
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    • 2013
  • Ambers have been used mostly as beads, jewelry and ornaments from ancient times and excavated as a buried artifact. When excavated, they are severely weathered to be cracked, exfoliated and disintegrated. Monitoring of changes in composition of amber according to weathering is very important for diagnosing the condition of amber and applying conservation materials and techniques. In this study, we tried to find the components of amber by analyzing amber with pyrolysis/GC/MS. The changes in the composition of pyrolzates after artificial ageing for 60 days under heat and oxygen were also observed. Abietic acid was detected as a main component of fresh amber and monoterpene, alkene, aromatic hydrocarbon were detected as major pyrolyzates. Changes with artificial ageing was estimated by comparing the peak area ratio of 23 components, and it was found that abietic acid abruptly decreased in the presence of heat and oxygen together, revealing that oxygen is a key factor to the deterioration of amber. It was also tried to understand the weathered surface of original amber gemstone based on the result of this ageing experiment.

Establishment of a Selection System for the Site-Specific Incorporation of Unnatural Amino Acids into Protein (비천연 아미노산의 위치특이적 단백질 삽입을 위한 Amino Acyl-tRNA Synthetase 선별시스템 개발)

  • Edan, Dawood Salim;Choi, Inkyung;Park, Jungchan
    • Korean Journal of Microbiology
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    • v.50 no.1
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    • pp.1-7
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    • 2014
  • Site-specific incorporation of unnatural amino acids (SSIUA) into protein can be achieved in vivo by coexpression of an orthogonal pair of suppressor tRNA and engineered aminoacyl-tRNA synthetase (ARS) that specifically ligates an unnatural amino acid to the suppressor tRNA. As a step to develop the SSIUA technique in Escherichia coli, here we established a new 2-step screening system that can be used for selecting an ARS variant(s) that ligates an unnatural amino acid to a suppressor tRNA. A positive selection system consists of chloramphenicol acetyl transferase gene containing an amber mutation at the $27^{th}$ residue, and efficiently concentrated amber suppressible ARS with a maximum enrichment factor of $9.0{\times}10^5$. On the other hand, a negative selection system was constructed by adding multiple amber codons in front of a lethal gene encoding the control of cell death B toxin (ccdB) which acts as an inhibitory protein of bacterial topoisomerase II. Amber suppression of ccdB by an orthogonal pair of Saccharomyces cerevisiae tyrosyl-tRNA synthetase (TyrRS) and an amber suppressor tRNA significantly inhibits bacterial growth. This selection system was also able to efficiently remove amber suppressible ARS which could ligate natural amino acids to the suppressor tRNA. Thus, sequential combination of these two selection systems might be able to function as a powerful tool for selecting an ARS variant that specifically ligates an unnatural amino acid to the suppressor tRNA from an ARS mutant pool.

A Study of Conservation Treatment of the Amber Relic of the Buddha Excavated from the Naksan Temple (낙산사 출토 호박사리호 보존처리에 관한 연구)

  • Ham, Chul-Hee;Kang, So-Yeong;Hwang, Jin-Ju
    • 보존과학연구
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    • s.30
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    • pp.21-29
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    • 2009
  • This study shows the research of causes of deterioration of the amber from foreign cases. It also presents the conservation treatment as well as tests to examine the effect of the organic solvents on the amber in order to find reversible and safe reinforcing agent which will prevent further damage of the artefact. The result showed that there was no noticeable change in the weight of the artefact as well as on the appearance of the surface when Mineral spirit(White spirit), which is aliphatic hydrocarbon group, and Xylene, aromatic hydrocarbons group was applied. Reinforcement and restoration was carried out using Paraloid(R) B67, which has a broad option for the solvent, in Xylene. Finally, it is recommended that the use of polar organic solvent in the conservation treatment of amber artefacts should be limited.

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Analysis of ambers with different origin by IR and py/GC/MS (산지별 호박(amber)의 IR 및 py/GC/MS 특성 분석)

  • Park, Jong-Seo;Lim, Yu-Jin
    • Analytical Science and Technology
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    • v.24 no.4
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    • pp.256-265
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    • 2011
  • Ambers have been used as a gemstone and a religious object since the ancient times and found in several archaeological sites in Korea. To prepare an enhanced conservation measures, we surveyed the chemical and spectroscopic properties of the ambers according to the provenance. Total 14 amber samples were collected from 6 different provenances including Baltic, Chiapas, Colombian, Dominican, Fushun and Madagascar amber. Infrared (IR) spectroscopic analysis was conducted for the non-destructive examination of the amber samples. They were also analyzed with pyrolysis/GC/MS (py/GC/MS) at the pyrolysis temperature of $300^{\circ}C$ with the on-line derivatization to trimethylsilyl ester. Baltic shoulder corresponding to the absorption at $1250cm^{-1}{\sim}1150cm^{-1}$ appeared in the IR spectrum of Baltic amber. IR spectra of the other ambers also showed somewhat distinctive characteristic peaks. In py/GC/MS analysis peaks assignable to succinic acid, dehydroabietic acid and pimaric acid were detected, which are known to be the components of the amber. In the meanwhile, the presence of compounds appearing in certain amber will be applied to differentiate the provenances of amber relics if their fragments are available for the analysis. These results are expected to help the confirmation of archaeological amber relics and archaeometric interpretation of provenances and manufacturing techniques.

Change of fluorescence in ambers according to artificial aging (인공열화에 따른 호박(amber)의 형광특성 변화)

  • Park, Jong-Seo;Lim, Yu-Jin
    • Analytical Science and Technology
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    • v.25 no.3
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    • pp.197-206
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    • 2012
  • Ambers are composed of polymer molecules which contain aromatic moieties such as benzene, naphthalene, phenanthrene and anthracene. They emit fluorescence when irradiated with ultraviolet light, which was used for confirming an amber. The fluorescence of amber, however, tends to decrease as the surface of amber is weathered with light, heat, oxygen for a long time. In this study, the reliability of confirming amber with its fluorescence by measuring the changes of fluorescence after artificial aging. Aging factors were UV light (${\lambda}$=340 nm), oxygen with heat (100%, $90^{\circ}C$) and heat ($90^{\circ}C$) and aging time was for 5, 15, 30 and 60 days, respectively. In the excitation and emission spectra of amber, the intensity decreased and the maximal wavelength was shifted to longer wavelength with artificial aging time. Especially, there was a drastic decrease in the intensity of spectra to 1.7% of initial value after 60 days aging under oxygen with heat. Only in Colombian amber there showed an increase of fluorescence intensity for a certain aging time, which could be explained by the production of aromatic ring in the presence of light and heat. Conclusively, the fluorescence can be lessened by the natural weathering with light, heat and oxygen and it is not accurate to recognize amber just with UV irradiation method.

An Amber Force Field for S-Nitrosoethanethiol That Is Transferable to S-Nitrosocysteine

  • Han, Sang-Hwa
    • Bulletin of the Korean Chemical Society
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    • v.31 no.10
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    • pp.2903-2908
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    • 2010
  • Protein S-nitrosation is common in cells under nitrosative stress. In order to model proteins with S-nitrosocysteine (CysSNO) residues, we first developed an Amber force field for S-nitrosoethanethiol (EtSNO) and then transferred it to CysSNO. Partial atomic charges for EtSNO and CysSNO were obtained by a restrained electrostatic potential approach to be compatible with the Amber-99 force field. The force field parameters for bonds and angles in EtSNO were obtained from a generalized Amber force field (GAFF) by running the Antechamber module of the Amber software package. The GAFF parameters for the CC-SN and CS-NO dihedrals were not accurate and thus determined anew. The CC-SN and CS-NO torsional energy profiles of EtSNO were calculated quantum mechanically at the level of B3LYP/cc-pVTZ//HF/6-$31G^*$. Torsional force constants were obtained by fitting the theoretical torsional energies with those obtained from molecular mechanics energy minimization. These parameters for EtSNO reproduced, to a reasonable accuracy, the corresponding torsional energy profiles of the capped tripeptide ACE-CysSNO-NME as well as their structures obtained from quantum mechanical geometry optimization. A molecular dynamics simulation of myoglobin with a CysSNO residue produced a well-behaved trajectory demonstrating that the parameters may be used in modeling other S-nitrosated proteins.

Characterization of Sun Spangle Formation in the Transparent Baltic Amber by Heat Treatment (열처리를 통한 발틱 호박의 Sun spangle생성에 관한 연구)

  • Chung, Hyo-Jin;Seo, Jin-Gyo;Park, Jong-Wan
    • Journal of the Mineralogical Society of Korea
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    • v.22 no.4
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    • pp.395-405
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    • 2009
  • The maturity of amber matrix and inclusion was studied to explore the effect of heat treatment on the formation of the sun spangle in the transparent Baltic amber from Gdansk, Poland. Optical microscope revealed two types of inclusions in the original amber samples. The inclusions in the amber which had sun spangle were dramatically expanded by the heat treatment, comparing to those without sun spangle which contains fissure and shows different refractive index. The amber sample which didn't show sun spangle after the heat treatment showed a strong carbon related peak in TOF-SIMS spectra and weak oxygen related peak. it means that the maturity of this amber sample is comparatively higher than the one with spangle. The two amber samples show similar IR spectra before the heat treatment. However, the amber which had sun spangle exhibited an additional 1019 $cm^{-1}$ absorption peak and a more intense 887 $cm^{-1}$ peak. The different chemical compositions between the two types of amber is believed to be due to the different absorption spectra between 1000~600 $cm^{-1}$. According to the study of DSC analysis, the two samples show different DSC profiles. Although they have the same geological origin, their geological ages are different and have different chemical compositions. Thus, they exhibited different behavior after the heat treatment. The formation of sun spangle seemed to depend on the difference in the heat expansion rate of amber matrix with different maturity and chemical compositions.

흉부의 ANALOG-DIGITAL 영상에 대한 비교 평가

  • Jang, Yeong-Ae;Mun, Hui-Seok;Im, Han-Yeong;O, Hyeon-Ju;Kim, Seong-Su;Kim, Yeong-Il
    • Korean Journal of Digital Imaging in Medicine
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    • v.2 no.1
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    • pp.74-82
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    • 1996
  • This paper were studied the evaluation in compare with the conventional and AMBER of analog images, PACS and CR of digital images which were collected every ten sampling chest images with the J.J.Vucicuh chest evaluated chart, and were measured the chest phantom surface dose and the density of several part in chest images. The evaluated numbers were total 22 persons who were 6 persons of the M.D., 6 of the radiotechnological professors and 10 of the radiotechnologists. The obtained results summarized as following : 1. Approaching the optimum standard density of the several part in chest images drew near at the lung round region density in PACS images, the sternum region density in CR and AMBER images, the heart region density in CR AMBER images, the diaphram region density in AMBER and conventional images. 2. The evaluation measured surface dose were appeared orderly lesser dose at the AMBER images (spine 21 mR, lung 2mR, heart 12mR, apex 6mR) than the conventional images(32 mR), CR images(38mR) and PACS images(81mR). 3. The anatomical physical evaluation marks were taken the highest points at CR images(88.3), and orderly PACS images(82), AMBER images(79.2) and conventional images(65.2). 4. It is exposured with lesser surface dose at the analog images, but analog images leaves much room for image quality improvement, and digital images demand for lesser exposure surface dose, although excellent image quality.

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