• Title/Summary/Keyword: Oligo(3

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Identification of Cuts-specific Myogenic Marker Genes in Hanwoo by DNA Microarray (DNA Microarray 분석을 통한 한우 부위별 특이 마커 유전자의 발굴)

  • Lee, Eun-Ju;Shin, Yu-Mi;Lee, Hyun-Jeong;Yoon, Du-Hak;Chun, Tae-Hoon;Lee, Yong-Seok;Choi, In-Ho
    • Journal of Animal Science and Technology
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    • v.52 no.4
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    • pp.329-336
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    • 2010
  • Myogenic satellite cells (MSCs) are mononuclear, multipotent progenitors of adult skeletal muscle possessing a capacity of forming adipocyte-like cells (ALC). To identify the skeletal muscle type-specific myogenic and adipogenic genes during MSCs differentiation, total RNA was extracted from bovine MSCs, myotube-formed cell (MFC), and ALC from each of Beef shank, Longissimus dorsi, Deep pectoral, and Semitendinosus. DNA microarray analysis (24,000 oligo chip) comparing MSCs with MFC and ALC, respectively, revealed 135 differentially expressed genes (> 4 fold) among four cuts. Real-time PCR confirmed expression of 29 genes. Furthermore, the whole tissue sample RNAs analysis showed 6 differentially expressed genes in Beef shank. Among which, 1 gene in MSCs, 4 in MFC, and 1 in ALCs were highly expressed. This study will provide an insight for better understanding the molecular mechanism of differentiation of skeletal muscle type-specific MSCs. The identified genes may be used as marker to distinguish skeletal muscle types.

Detection of Norovirus in Contaminated Ham by Reverse Transcriptase-PCR and Nested PCR

  • Kim, Seok-Ryel;Kim, Du-Woon;Kwon, Ki-Sung;Hwang, In-Gyun;Oh, Myung-Joo
    • Food Science and Biotechnology
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    • v.17 no.3
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    • pp.651-654
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    • 2008
  • In order to enhance the efficacy of norovirus detection by reverse transcriptase-polymerase chain reaction (RT-PCR) and nested PCR, this study developed a norovirus mRNA concentration method using poly oligo dT-conjugated magnetic beads. An efficient norovirus detection protocol was performed on commercial ham using 2 viral elution buffers (glycine buffer and Tris beef extract buffer) and 2 concentration solutions [polyethylene glycol (PEG) and zirconium hydroxide]. The different approaches were verified by RT-PCR and nested PCR. This method was performed on ham in less than 8 hr by artificial inoculation of serial dilutions of the virus ranging from 1,000 to 1 RT-PCR unit/mL. The viral extraction and concentration method had 10-fold higher sensitivity using the combination of Tris beef extract buffer and PEG as compared to glycine buffer and zirconium hydroxide. This method proved that RT-PCR and nested PCR have the sensitive ability to detect norovirus in commercial ham, in that norovirus was successfully detected in artificially contaminated samples at a detection level as low as 1-10 RT-PCR unit/mL. Overall, such a detection limit suggests this protocol is both quick and efficient in terms of its potential use for detecting norovirus in meat products.

Partial Purification and Properties of Non-specific $\beta$ -fructofuranosidase Produced by Bacillus subtilis (Bacillus subtilis가 생산하는 비특이적 $\beta$-fructofuransoidase의 부분정제 및 특성)

  • 송근섭;엄태붕
    • Microbiology and Biotechnology Letters
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    • v.18 no.5
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    • pp.484-489
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    • 1990
  • An intracellular inulase ( fJ-fructofuranoside fructohydrolase, EC 3.2.1.26) from Bacillus subtilis has been partially purified and its mode of action and general properties were studied. The enzyme had an apparent molecular weight of 49,000 as estimated by gel filtration and its pI point was 5.2. Substrate concentration studies showed an apparent Km of 10 mM for sucrose and of 18 mM for raffinose. The enzyme was an acid-labile protein with a pH optimum of 6.6. The optimum temperature was 50$^{\circ}$C. The enzyme acts on straight chain oligo- and poly-fructosides of the inulin series via a exo-wise cleavage mechanism, as well as on sucrose.

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Pathological and molecular comparisons of five distinct species of pepper-infecting Potyviruses (oral)

  • Yoon, H.I.;Chung, H.M.;Ryu, K.H.
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.113.2-114
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    • 2003
  • Five pepper-infecting potyviruses, Pepper mottle virus (PepMoV), Chilli veinal mottle virus (CVMV), Pepper veinal mottle virus (PVMV), Pepper severe mosaic virus (PSMV) and Tobacco each virus (TEV), are known filamentous virus and can be infected pepper crops systemically. To understand pathology and genome information of the five viruses on pepper plants, host reactions and sequences were compared to the 5 viruses. Five potyviruses were inoculated onto some typical cultivars of hot peppers and compared their symptoms, and virus accumulations. A set of degenerate primers for potyviruses were applied to 5 viruses and RT-PCR was performed. RT-PCR products containing partial nuclear inclusion b and coat protein (CP) genes were cloned. Then, oligo dT primer and species-specific primer were redesigned to amplify the C-terminal part of CP and 3' noncoding regions of each viruses. Sequences of the viruses were analyzed and compared to serological relationships among the viruses. The data can be useful for screening of potyviruses in pepper plants and pathogen-derived transgenic pepper plant development.

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Genes expression monitoring using cDNA microarray: Protocol and Application

  • Muramatsu Masa-aki
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2000.11a
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    • pp.31-41
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    • 2000
  • The major issue in the post genome sequencing era is determination of gene expression patterns in variety of biological systems. A microarray system is a powerful technology for analyzing the expression profile of thousands of genes at one experiment. In this study, we constructed cDNA microarray which carries 2,304 cDNAS derived from oligo-capped mouse cDNA library. Using this hand-made microarray we determined gene expression in various biological systems. To determine tissue specific genes, we compared Nine genes were highly-expressed in adult mouse brain compared to kidney, liver, and skeletal muscle. Tissue distribution analysis using DNA microarray extracted 9 genes that were predominantly expressed in the brain. A database search showed that five of the 9 genes, MBP, SC1, HiAT3, S100 protein-beta, and SNAP25, were previously known to be expressed at high level in the brain and in the nervous system. One gene was highly sequence similar to rat S-Rex-s/human NSP-C, suggesting that the gene is a mouse homologue. The remaining three genes did not match to known genes in the GenBank/EMBL database, indicating that these are novel genes highly-expressed in the brain. Our DNA microarray was also used to detect differentiation specific genes, hormone dependent genes, and transcription-factor-induced genes. We conclude that DNA microarray is an excellent tool for identifying differentially expressed genes.

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Bifidogenic Effect of Glucooligosaccharide Prepared from Glucose by Extrusion Process

  • Ahn, Jun-Bae;Hwang, Jae-Kwan;Kim, Chong-Tai;Lee, Ke-Ho;Park, Jong-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.7 no.3
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    • pp.174-179
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    • 1997
  • In order to investigate effect of glucooligosaccharide (GOS) prepared by extrusion process as a bifidogenic factor, cultivation of Bifidobacterium sp., Bacteroides fragilis and Clostridium perfringens was done and analyzed. B. fragilis and C. perfringens were able to utilize only 16% and 11% of the oligosaccharides in GOS, respectively, whilst Bifidobacterium sp. FBD-22 could utilize 38%. Especially, many kinds of oligo saccharides in GOS were able to be utilized selectively only by Bifidobacterium sp.. In case that GOS, as a carbon source, was used in the co-cultivation by Bifidobacterium sp., B. fragilis and C. perfringens, growth of Bifidobacterium sp. was not influenced by the existence of B. fragilis and C. perfringens. Bifidobacterium sp. showed advantage on carbon source competition for GOS with B. fragilis. Acetic acid, antimicrobial agent in the intestine, was produced two times more from GOS than glucose in co-cultures of three strains. Therefore, it is suggested that GOS can be a potent bifidogenic factor which proliferates the population of Bifidobacterium sp. and may finally improve the intestinal environments of human.

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Molecular Cloning of H-Y Antigen Gene III. Construction of Mouse Testis cDNA Library and Screening of H-Y Ag Gene (H-Y 항원 유전자의 클로닝에 관한 연구 III. 생쥐정소 cDNA Library 구성과 유전자의 검색)

  • 이정렬;김창규;김종배
    • Korean Journal of Animal Reproduction
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    • v.17 no.1
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    • pp.43-48
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    • 1993
  • These experiments were carried out to construct mouse testis cDNA library and to to seen H-Y Ag gene. Mouse testis was obtained from BALB/c inbreed mouse that was after-born 1 week. Isolation of mouse testis total RNA was carried out by guanidum/cesium choloride, poly(A+) mRNAs were purified by oligo d(T)-cellulose chromatography method. To investigate protein synthesis activity, in-vitro translation carried out by total RNA and poly(A+) mRNA. The products of in-vitro translation were identified in 12.5% PAGE. Single strand DNA and double strand DNA were synthesized from poly(A+) mRNA and purified using phenol/chloroform/isoamylalcohol. Synthesized cDNA was combined with cohesive Eco RI polylinker, its recombination efficiencies were identified by X-gal and IPTG. In the cDNA library, 1$\times$107 phagemids were screened with 32P labelled probe. Hybridization were carried on $65^{\circ}C$ for 16~20hours. And 1$\times$106 phagemids were screened with rabbit-anti-H-Y. In former, select 5 positive clones, and later, 1 positive clone. Its southern blot analysis showed various size of insert cDNA from 0.7kb to 3kb.

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Preparation of Polyesteramide from Oligobutylene Terephtahlate and 6-amino Caproic Acid (올리고 부틸렌 테레프탈레이트와 6-아미노 카프론산으로부터 포리에스테르 아미드의 합성)

  • Cho, Soon-Chae;Choi, Doo-Jin;Kim, Seon-Ho;Rhee, Johng-Moon;Pak, Pyong-Ki;Ahn, Jong-Yeol;Kim, Young-Gil
    • Textile Coloration and Finishing
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    • v.3 no.1
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    • pp.17-24
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    • 1991
  • Polyesteramide(PEA) has been synthesized with different molar ratio of 6-aminocaproic acid(ACA) and oligo butylene terephthalate(OBT) prepared from Dimethyl terephthalate(DMT) and 1, 4-butanediol(BD) by the melt polymerization. Oligomer and polymer molecular structure were characterized by IR spectra, H'-NMR spectra. Also the thermal properties were examined by thermogravimetric analysis(TGA) and differential scanning calorimetry(DSC). DSC and TGA results exhibited that the$ T_m$ s were detectable in the vicinity of $170-220^{\circ}C$ and the Ta's were detectable in the vicinity of $380-390^{\circ}C$ . X-ray diffraction analysis results show that the synthesized polyesteramide has high degree of crystallinity.

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Chitinase-producing Salinivibrio bacteria isolated from salt-fermented shrimp with antimicrobial and safety assessments

  • Le, Bao;Chung, Gyuhwa;Yang, Seung Hwan
    • Journal of Applied Biological Chemistry
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    • v.61 no.3
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    • pp.233-238
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    • 2018
  • Chitinases are glycosyl hydrolases which cleave the ${\beta}$-1,4 linkage of chitin into oligo or monomers of N-acetylglucosamine. These bacterial enzymes have been used for a wide range of applications in the food and pharmaceutical industries. In this study, we isolated two potential chitinolytic strains, BAO-01 and BAO-02, from salt-fermented shrimp, which were shown to belong to the genus Salinivibrio through genetic characterization using 16S rRNA. These isolates were gram-positive, rod-shaped, and non-spore forming. BAO-01 showed greater growth and chitinase activity than BAO-02 after the incubation at $37^{\circ}C$ for 4 days. Both strains grew on a wide range of carbon and nitrogen sources, pH values, temperatures, and salt levels. However, they showed minor biochemical differences. In addition, their antimicrobial activities against foodborne pathogens and antibiotic susceptibilities were evaluated. These Salinivibrio spp. did not show bioamine production, hemolytic activity, and mucin degradation. Therefore, the in vitro screening results suggested that these bacteria could be widely used as new candidates for chitin hydrolyzation and seafood fermentation.

Toxicogenomics Study on ${\alpha}-Naphthylisothiocyanate\;(ANIT)$ Induced Hepatotoxictiy in Mice

  • Hwang, Ji-Yoon;Lim, Jung-Sun;Jeong, Sun-Young;Park, Han-Jin;Cho, Jae-Woo;Yoon, Seok-Joo
    • Molecular & Cellular Toxicology
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    • v.2 no.1
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    • pp.48-53
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    • 2006
  • [ ${\alpha}-Naphthylisothiocyanate$ ] (ANIT) induces intrahepatic cholestasis, involving damage to biliary epitheial cells. This study investigates hepatic gene expression and histopathological alterations in response to ANIT treatment in order to elucidate early time response of ANIT-induced hepatotoxicity. ANIT was treated with single dose (3, 6, and 60 mg/kg) in corn oil by oral gavage. Serum biochemical and histopathological observation were performed for evaluation of hepatotoxicity level. Affymetrix oligo DNA chips were used for gene expression profile by ANIT-induced hetpatoxicity. Hepatic enzyme levels (ALT, AST, and ALP) were increased in 24 hr high dose group. In microscopic observations, moderate hepatocellular necrosis, were confirmed 24 hr high dose groups. We found that gene expression patterns were dependent on time and dose. Our selected genes were related inflammation and immunomodulation. In this study, ANIT-induced hepatotoxicity was involved in acute phase responses and provides evidence for role of neutrophil could be mechanism associated with ANIT-mediated hepatotoxicity.