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Cloning of Cytochrome P450 Gene involved in the Pathway of Capsidiol Biosynthesis in Red Pepper Cells (고추세포에서 Capsidiol 생합성을 유도하는 Cytochrome P450 유전자의 탐색)

  • Kwon, Soon-Tae;Kim, Jae-Sung;Jung, Do-Cheul;Jeong, Jeong-Hag;Hwang, Jae-Moon;Oh, Sei-Myoung
    • Journal of Life Science
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    • v.13 no.6
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    • pp.879-888
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    • 2003
  • In order to measure the enzyme activity of 5-epi-aristolochene hydroxylase, one of cytochrome P450 (P450) enzymes in eicitor-treated pepper cell, we used in vivo assay method and demonstrated a dramatic suppression of the activity by P450-inhibitors, ancymidol and ketocornazole. Using RT-PCR method with degenerate primer of the well conserved domains found within most P450-enzymes, and using cDNA library screening method, one distinct cDNA, being designated P450Hy01, was successfully isolated from elicitor-treated pepper cells. P450Hy01 mRNA was all induced in elicitor-treated cells whereas never induced in control cells. Moreover, levels of P450Hy01 expression were highly correlated with the levels of extracellular capsidiol production by different elicitors in cell cultures. P450Hy01 transcript was also induced by several other elicitors such as, cellulase, arachidonic acid, jasmonic acid, yeast extract as well as UV stress. P450Hy01 sequence contained high probability amino acid matches to known Plant P450 genes and ORF with a conserved FxxGxRxCxG heme-binding domain. P450Hy01 cDNA showed 98% of homology in sequence of nucleotide as well as amino acid to 5-epi-aristolochene-1, 3-hydroxylase (5EAl, 3H) which has been isolated in tobacco cells, suggesting that P450Hy01 is prominent candidate gene for P450-enzyme encoding 5EAl, 3H in pepper cell.

Use of In Vivo-Induced Antigen Technology to Identify In Vivo-Expressed Genes of Campylobacter jejuni During Human Infection

  • Hu, Yuanqing;Huang, Jinlin;Li, Qiuchun;Shang, Yuwei;Ren, Fangzhe;Jiao, Yang;Liu, Zhicheng;Pan, Zhiming;Jiao, Xin-An
    • Journal of Microbiology and Biotechnology
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    • v.24 no.3
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    • pp.363-370
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    • 2014
  • Campylobacter jejuni is a prevalent foodborne pathogen worldwide. Human infection by C. jejuni primarily arises from contaminated poultry meats. Genes expressed in vivo may play an important role in the pathogenicity of C. jejuni. We applied an immunoscreening method, in vivo-induced antigen technology (IVIAT), to identify in vivo-induced genes during human infection by C. jejuni. An inducible expression library of genomic proteins was constructed from sequenced C. jejuni NCTC 11168 and was then screened using adsorbed, pooled human sera obtained from clinical patients. We successfully identified 24 unique genes expressed in vivo. These genes were implicated in metabolism, molecular biosynthesis, genetic information processing, transport, and other processes. We selected six genes with different functions to compare their expression levels in vivo and in vitro using real-time RT-PCR. The results showed that the selected six genes were significantly upregulated in vivo but not in vitro. In short, these identified in vivo-induced genes may contribute to human infection of C. jejuni, some of which may be meaningful vaccine candidate antigens or diagnosis serologic markers for campylobacteriosis. IVIAT may present a significant and efficient method for understanding the pathogenicity mechanism of Campylobacter and for finding targets for its prevention and control.

Cataloguing of Anther Expressed Genes through Differential Slot Blot in Oriental Lily (Lilium Oriental Hybrid 'Acapulco') (아카풀코나리에서 Differential Slot Blot을 이용한 약발현 유전자 목록작성)

  • Suh, Eun-Jung;Yu, Hee Ju;Han, Bong Hee;Lim, Yong Pyo;Jeong, Mi-Jeong;Lee, Seong-Kon;Kim, Dong-Hern;Chang, An-Cheol;Yae, Byeong Woo
    • Horticultural Science & Technology
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    • v.31 no.5
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    • pp.598-606
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    • 2013
  • Anther is the major organ of flower in responsible to reproduction and outward appearance. From anther-specific cDNA library of Lilium Oriental Hybrid 'Acapulco', 2000 expressed sequence tags were selected randomly. Differential slot blot analysis with cDNA probes from the anther and leaf was used to get anther-expressed clone and 570 non-redundant ESTs were obtained and sequenced. Compared to the GenBank database using BLASTX algorithm, 191 clones showed significant similarity but others (66.5%) did not measured to known sequence. Functional categories according to gene ontology (GO) annotation included sequence representing a significant portion of protein in cell and cell part respectively. A transcriptional analysis at 7 different organs and developmental stage was performed using northern blot with thirty ESTs as putative anther specific gene. This report suggest that selection of anther expressed clone using differential slot blot was considered as very effective tool and our current study can provide fundamental information on the lily anther including pollen furthermore.

Isolation and Characterization of Helicobacter pylori Urease Inhibitor from Rubus coreanus Miquel (복분자(Rubus coreanus Miquel)로부터 Helicobacter pylori Urease Inhibitor의 분리 및 특성)

  • 양성우;호진녕;이유현;신동훈;홍범식;조홍연
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.5
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    • pp.769-777
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    • 2004
  • A Helicobacter pylori urease inhibitor from Rubus coreanus Miquel has been isolated and partially characterized for aiming to Prevent H. pylori growth and decrease harmful accumulation of ammonia in human gastric mucosa. We screened urease inhibitory activities in 519 extracts library prepared by solvent extraction from 173 kinds of edible plants, medicinal herbs, herbs and seaweeds using a colorimetric urease assay system. As results of primary and secondary screening, 70% acetone extract of Rubus coreanus Miquel was selected as potent candidate, showing about 24% inhibitory activity. The acetone extract was sequentially partitioned into RCE/RCWI and RCB/RCW2 layers with ethyl acetate and butanol. The major active component in RCW2, water layer from butanol fractionation was revealed to be peptidic or proteinous substance by inhibitory activity determination after pronase digestion and periodate oxidation. RCW2-IIIc a was isolated by sequential column chromatography on DEAE-Toyopearl 650C, Butrl-Toyopearl 650M and Sephadex LH-20. The isolated urease inhibitor RCW2-IIIc $\alpha$, was highly pure proteinous substance with molecular weight of 13kDa by high-performance gel permeation liquid chromatography. RCW2-IIIc$\alpha$ has about 5 times higher inhibitory activity than 70% acetone extract, showing high stability against heat treatment and peptic digestion.

Effects of Tumor Necrosis Factor-alpha Inhibitors on the Incidence of Tuberculosis (Tumor Necrosis Factor-alpha 저해제가 결핵 발생에 미치는 영향)

  • Park, Hyun Jin;Choi, Bo Yoon;Sohn, Minji;Han, Na Young;Kim, In-Wha;Oh, Jung Mi
    • Korean Journal of Clinical Pharmacy
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    • v.28 no.4
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    • pp.333-341
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    • 2018
  • Objective: Tumor necrosis factor-alpha (TNF-alpha) inhibitors are used as a treatment in various immune-mediated inflammatory diseases (IMIDs). Tuberculosis (TB) risk is reported in several meta-analyses in patients treated with TNF-alpha inhibitors. The purpose of this study is to collect, review, and evaluate the TB risk in TNF-alpha inhibitors according to IMIDs indications and between soluble-receptor TNF-alpha inhibitor and monoclonal-antibody TNF-alpha inhibitors. Methods: A systematic literature search on systematic reviews and meta-analyses was performed in PubMed, MEDLINE, Cochrane library, and EMBASE. We identified meta-analyses that evaluated TB infection risk of TNF-alpha inhibitors in IMIDs patients. Results: Thirteen meta-analyses including 41 study results were included in this umbrella review. IMIDs patients treated with TNF-alpha inhibitors had an increased risk of TB than control group (placebo with or without standard therapy patients) (relative risk ratio (RR) 2.057, 95% confidence interval (CI) 1.697 to 2.495). Among them, RA patients with TNF-alpha inhibitors had a higher risk of TB than control group (RR 1.847, 95% CI 1.385 to 2.464), and non-RA patients with TNF-alpha inhibitors had an increased risk of TB (RR 2.236, 95% CI 1.284 to 3.894). In subgroup analysis on TB risk between soluble-receptor TNF-alpha inhibitor and monoclonal-antibody TNF-alpha inhibitors in RA patients, the analysis indicated that monoclonal-antibody TNF-alpha inhibitors had higher risk of TB than soluble-receptor TNF-alpha inhibitor (RR 2.880, 95% CI 1.730 to 4.792). Conclusion: This umbrella review confirms that the risk of TB is significantly increased in TNF-alpha inhibitor treated patients compared to control group.

A Textual Research on Hu ShunShen (胡舜申)'s Life and Works (호순신(胡舜申)의 생애와 저술에 관한 연구)

  • Oh, Dong Kee
    • Korean Journal of Heritage: History & Science
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    • v.44 no.3
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    • pp.44-61
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    • 2011
  • This study consider the life and works of Hu ShunShen(胡舜申) who was the author of JiRiSinBub(地理新法) which the representative FengShui book in Choson dynasty. His adult name is RuJia(汝嘉). He was born in on September 6, 1091 at JiXi(績溪) in China as a son of Ho Xian(胡咸). He left his hometown with his family to avoid war and settled down in HuZhou(胡州). He took up an official post with his brother's YinPu(蔭補), and held several provincial official posts. After serving as vice governor(通判) of ShuZhou(舒州), he became supervisor of taoist temple(崇道觀) in TaiZhou(台州) and retired from office. After burying his father, he took an interest in fengShui(風水) and studied for a long time. People say that JiangXiDilixinfa(江西地理新法) is the well-known FengShui book written by him. When he was 74 years old, he suggested opening SheMen(蛇門) gate and XuMen(胥門) gate in SuZhou(蘇州) castle by "WoMenZhongGao(吳門忠告)". But it didn't come ture. He died March 9, 1177 at the age of 87 and was buried in HuZhou(胡州). His elder brother Hu Shunzhi(胡舜陟) and nephew HuZi(胡仔) is well-known. He had a son named Hu wei(胡偉) who served pacification commissioners of JiangXi(江西宣撫使). His Works were YiSiSiZhouLu(乙巳泗州錄), YiYouBiLuanLu(己酉避亂錄) as essay and YinYangBeiYong(陰陽備用), JiRiSinBub(地理新法), "WoMenZhongGao(吳門忠告)" as fengShui text.

Some Properties and Microbial Community Changes of Gul (Oyster) Jeotgal during Fermentation

  • Kim, Jeong A;Yao, Zhuang;Kim, Hyun-Jin;Kim, Jeong Hwan
    • Microbiology and Biotechnology Letters
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    • v.47 no.3
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    • pp.343-349
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    • 2019
  • Gul jeotgals (GJs) were prepared using solar salt aged for 3 years. One sample was fermented using starters, such as Bacillus subtilis JS2 and Tetragenococcus halophilus BS2-36 (each $10^6CFU/g$), and another sample was fermented without starters for 49 days at $10^{\circ}C$. Initial counts of bacilli and lactic acid bacteria (LAB) in non-starter GJ were found to be $3.20{\times}10^2$ and $7.67{\times}10^1CFU/g$ on day 0, and increased to $1.37{\times}10^3$ and $1.64{\times}10^6CFU/g$ on day 49. Those of starter GJ were found to be $2.10{\times}10^5$ and $3.30{\times}10^7CFU/g$ on day 49, indicating the growth of starters. The pH values of GJ were $5.93{\pm}0.01$ (non-starter) and $5.92{\pm}0.01$ (starter) on day 0 and decreased to $5.78{\pm}0.01$ (non-starter) and $5.75{\pm}0.01$ (starter) on day 49. Amino-type nitrogen (ANN) production increased continuously during fermentation, and $407.19{\pm}15.85$ (non-starter) and $398.04{\pm}13.73$ (starter) mg% on day 49. Clone libraries of 16S rRNA genes were constructed from total DNA extracted from non-starter GJ on days 7, 21, and 42. Nucleotide sequences of Escherichia coli transformants harboring recombinant pGEM-T easy plasmid containing 16S rRNA gene inserts from different bacterial species were analyzed using BLAST. Uncultured bacterium was the most dominant group and Gram - bacteria such as Acidovorax sp., Afipia sp., and Variovorax sp. were the second dominant group. Bacillus amyloliquefaciens (day 7), Bacillus velezensis (day 21 and 42), and Bacillus subtilis (day 42) were observed, but no lactic acid bacteria were detected. Acidovorax and Variovorax species might play some role in GJ fermentation. Further studies on these bacteria are necessary.

Melanogenesis Promotion by 3-Deazaneplanocin A, a Specific Inhibitor of S-Adenosylhomocysteine Hydrolase, in B16/F10 Melanoma Cells (B16/F10 흑색종 세포에서 S-Adenosylhomocysteine Hydrolase 의 선택적 저해제 3-Deazaneplanocin A 에 의한)

  • Hwang, Yun Jeong;Boo, Yong Chool
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.47 no.2
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    • pp.107-121
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    • 2021
  • Skin hypopigmentation, which is observed in albinism or vitiligo, occurs when melanin synthesis is decreased by genetic, epigenetic, and other factors. To identify drug candidates that can promote melanin synthesis in cells, we screened an epigenetic modulator library consisting of 141 cell-permeable, small molecule drugs. B16/F10 murine melanoma cells were treated with each drug at 0.1 𝜇M and melanin synthesis and cell viability were subsequently monitored. As a result, (-)-neplanocin A, 3-deazaneplanocin A (DZNep), and DZNep hydrochloride were found to increase cellular melanin synthesis without causing cytotoxicity. Because these three structurally related drugs exhibited similar dose-dependent effects on melanin synthesis and cell viability, DZNep was selected as a representative drug for additional experiments. DZNep increased intracellular melanin content and tyrosinase (TYR) activity. DZNep also induced the expression of TYR, tyrosinase-related protein 1 (TYRP1), and dopachrome tautomerase (DCT) at the mRNA and protein levels. DZNep also induced the mRNA and protein expression of microphthalmia-associated transcription factor (MITF), a key regulator of melanin synthesis. DZNep is a specific inhibitor of S-adenosylhomocysteine hydrolase and it caused the accumulation of S-adenosylhomocysteine that inhibits histone methyltransferases in cells. This study suggests that melanogenesis can be modulated by targeting S-adenosylhomocysteine hydrolase in certain cellular contexts.

Framework Construction with Multimedia Component Management System on CORBA (CORBA 환경에서 멀티미디어 컴퍼넌트 관리 시스템을 통한 프레임워크 구축)

  • 김행곤
    • Journal of Korea Multimedia Society
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    • v.2 no.2
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    • pp.217-229
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    • 1999
  • Framework is the set of interrelated classes, constructing reusable design in specific domain or set of abstracted classes, and defines common architecture among applications included in domain. Developers can reuse not only class code but also wide range of knowledge on domain by reusing framework. In this papers, we present COM(Component-Oriented Methodology) for the reuse of framework, and develop construction environment for framework and domain development. That is, domain is analyzed by input of domain knowledge on real world to create software based on component, and hotspot is identified through analyzed information, and redesigned(refactoring) by putting additional information on users and developers. After that, I will create domain framework and application framework depending on domain. In this Component-oriented methodology, information is searched, understood and extracted or composite through component-pattern library storage internally. Then this information is classified into the information on component and pattern respectively, and used as additional information in redesigning. With this, developer can obtain reusability, easiness and portability by constructing infrastructure environment that allow to register, update and delete component through Component Pattern Management System(CPMS) under the development environment which can be easily applied to his own application using multimedia component, in this thesis, CORBA(Common Object Request Broker Architecture) environment.

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A design and implementation of Face Detection hardware (얼굴 검출을 위한 SoC 하드웨어 구현 및 검증)

  • Lee, Su-Hyun;Jeong, Yong-Jin
    • Journal of the Institute of Electronics Engineers of Korea SD
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    • v.44 no.4
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    • pp.43-54
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    • 2007
  • This paper presents design and verification of a face detection hardware for real time application. Face detection algorithm detects rough face position based on already acquired feature parameter data. The hardware is composed of five main modules: Integral Image Calculator, Feature Coordinate Calculator, Feature Difference Calculator, Cascade Calculator, and Window Detection. It also includes on-chip Integral Image memory and Feature Parameter memory. The face detection hardware was verified by using S3C2440A CPU of Samsung Electronics, Virtex4LX100 FPGA of Xilinx, and a CCD Camera module. Our design uses 3,251 LUTs of Xilinx FPGA and takes about 1.96${\sim}$0.13 sec for face detection depending on sliding-window step size, when synthesized for Virtex4LX100 FPGA. When synthesized on Magnachip 0.25um ASIC library, it uses about 410,000 gates (Combinational area about 345,000 gates, Noncombinational area about 65,000 gates) and takes less than 0.5 sec for face realtime detection. This size and performance shows that it is adequate to use for embedded system applications. It has been fabricated as a real chip as a part of XF1201 chip and proven to work.