• 제목/요약/키워드: Nucleotides

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Evolution of a dextransucrase gene for constitutive and hyper-production and for synthesis of new structure dextran

  • 강희경;김도만;장석상
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2003년도 생물공학의 동향(XII)
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    • pp.545-549
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    • 2003
  • After irradiation of a cloned dextransucrase gene (dsrB742) with ultrasoft X-ray, an E. coli transformant (pDSRB742CK) was first developed for the expression of an extracellular dextransucrase, having increased activity and the synthesis of a highly branched dextran. Seven nucleotides of the parent gene (dsrB742) were changed in the nucleotide sequences of dsrB742ck. Among them, four nucleotides were changed at the ORF of dsrB742, resulting in a 30 amino acids deletion in the N-terminal of DSRB742 dextransucrase. The activity of DSRB742CK dextransucrase in culture supernatant was approximately 2.6 times higher (0.035 IU/ml) than that of the DSRB742 clone. The pDSRB742CK clone produced DSRB742CK dextransucrase when grown both on a sucrose medium (inducibly) and on a glucose medium (constitutively). The DSRB742 clone did not produce dextran constitutively on a glucose medium. DSRB742CK dextran had 15.6% branching and 2.7-times higher resistance to dextranase hydrolysis compared to DSRB742 dextran. $^{13}C-NMR$ showed that DSRB742CK dextran contained ${\alpha}-(1{\rightarrow}3)$ branch linkages that were not present in DSRB742 dextran.

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저염 오징어 젓갈의 숙성에 따른 휘발성염기질소 및 유리 아미노산의 변화 (Changes of the Volatile Basic Nitrogen and Free Amino Acids according to the Fermentation of Low Salt Fermented Squid)

  • 오성천;조정순;남혜영
    • 한국식품조리과학회지
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    • 제16권2호
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    • pp.173-181
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    • 2000
  • To understand the influences of NaCl concentration and fermentation temperature on the ripening process of low salt fermented squids, squid with 5%, 7% and 9% salt were fermented at 10$\^{C}$ and 20$\^{C}$. The result of the changes of volatile basic nitrogen and free amino acids during the fermentation of squids are as follows. As a result of the observations on the changes of physicochemical components during the fermentation process of the low-salted squids, all the pH, VBN and NH$_2$-N were increased and therefore the fermentation was promoted. Considering the changes of net components according to the fermentation, ATP (Adenosine triphosphate) and ADP (Adenosine diphosphate) lost and could not be detected among the nucleotides and their related compounds. Besides, AMP (Adenosine monophosphate) existed only in the initial stage and inosine, hypoxanthine were the main components of nucleotides and their related compounds. Nonvolatile organic acids are mainly lactic acid, acetic acid and also they occupied more than 80%. Seeing the composition of free amino acid, the major amino acids are proline, arginine, methionine, alanine and glutamic acid.

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Genomic Structure of the Cu/Zn Superoxide Dismutase(SOD1) Gene from the Entomopathogenic Fungus, Cordyceps pruinosa

  • Park, Nam Sook;Jin, Byung Rae;Lee, Sang Mong
    • International Journal of Industrial Entomology and Biomaterials
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    • 제39권2호
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    • pp.67-73
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    • 2019
  • The genomic structure of the Cu/Zn superoxide dismutase (SOD1) gene from the entomopathogenic fungus, Cordyceps pruinosa was characterized. The SOD1 gene of C. pruinosa spans 947 nucleotides and consisted of four exons encoding for 154 amino acids and three introns. Four exons of the SOD1 gene are composed of 13, 331, 97 and 20 nucleotides respectively. Homology search of amino acid sequences of the SOD1 gene of C. pruinosa with another 13 fungi species showed higher sequence similarity of 69% ~ 95% and had the most highest sequence identity of 95% with Beauveria bassiana and Cordyceps militaris, which can easely infect domesticated Bombyx mori and another wild lepidopteran species in artificial or natual manner of infection. This SOD1 gene sequence showed copper, zinc and beta-barrel fold sites. Homology search showed that the Cu/Zn SOD1 gene from the entomopathogenic fungus, C. pruinosa is an orthologous gene homolog present in different species of organism whose ancestor predates the split between the relating species. In addition, C. pruinosa SOD1 gene is placed together within the ascomycetes group of fungal clade. From these results it is concluded that C. pruinosa SOD1 gene is orthologous gene having the same or very similar functions with a common evolutionary ancestor.

Quantitative Assay for the Binding of Jun-Fos Dimer and Activator Protein-1 Site

  • Lee, Sang-Kyou;Park, Se-Yeon;Jun, Gyo;Hahm, Eun-Ryeong;Lee, Dug-Keun;Yang, Chul-Hak
    • BMB Reports
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    • 제32권6호
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    • pp.594-598
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    • 1999
  • The Jun and Fos families of eukaryotic transcription factors form heterodimers capable of binding to their cognate DNA enhancer elements. We are interested in searching for inhibitors or antagonists of the binding of the Jun-Fos heterodimer to the activator protein-1 (AP-1) site. The basic-region leucine zipper (bZIP) domain of c-Fos was expressed as a fusion protein with glutathione S-transferase, and allowed to form a heterodimer with the bZIP domain of c-Jun. The heterodimer was bound to glutathione-agarose, to which were added radiolabeled AP-1 nucleotides. After thorough washing, the gel-bound radioactivity was counted. The assay is faster than the coventional electrophoretic mobility shift assay because the gel electrophoresis step and the autoradiography step are eliminated. Moreover, the assay is very sensitive, allowing the detection of picomolar quantities of nucleotides, and is not affected by up to 50% dimethylsulfoxide, a solvent for hydrophobic inhibitors. Curcumin and dihydroguaiaretic acid, recently known inhibitors of Jun-Fos-DNA complex formation, were applied to this Jun-GST-fused Fos system and revealed to decrease the dimer-DNA binding.

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흰쥐 신조직내 Cyclic Nucleotide 함량에 미치는 Hydrocortisone과 Furosemide의 영향 (Effect of Hydrocortisone and Furosemide on the Renal Cyclic Nucleotides Content in Rat)

  • 조규철;김인순;양재하;박영서
    • 대한약리학회지
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    • 제19권1호
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    • pp.93-99
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    • 1983
  • Hydrocortisone 5 mg/kg which exerts minimal effect on the renal function and furosemide 1 mg/kg which induces moderate amount of diuresis were injected intraperitoneally to study their effects on the renal cyclic nucleotides content in rats. 1) The renal tissue levels of cAMP were significantly increased by administration of hydrocortisone, but there was no significant change in the furosemide group compared with that of saline treated control group. Moderate elevation in renal cAMP level was noted by the combined administration of hydrocortisone and furosemide, but this elevation was less than that of hvdrocortisone treated group. 2) The renal cGMP level did not show nay remarkable change after the administration of hydrocortisone, however, there were a significant increase by the administration of furosemide alone or combination of both drugs. The level of renal cGMP was higher and maintained longer in the combined treated group than furosemide treated group. The result of this experiment indicates that the potentiating effect of hydrocortisone on the diuretic action of furosemide nay be related to the renal levels of cGMP rather than that of cAMP.

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Newly Designed Six-membered Azasugar-Containing Phosphorothioate Oligonucleotide as a Potent AIDS Therapeutic Drug

  • Bae, Yong-Soo
    • 대한약학회:학술대회논문집
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    • 대한약학회 2002년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2
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    • pp.155-160
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    • 2002
  • A series of modified oligonucleotides containing a phosphorothioate (P=S) backbone and a six-membered azasugar (6-AZS) as a sugar substitute in a nucleotide were synthesized and tested for their ability to inhibit the human immunodeficiency virus type I(HIV-l) in vitro without the aid of any transfecting agents. While P=S oligonucleotides with natural nucleotides had little anti-HIV-l activity, the six-membered azasugar nucleotide (6-AZN)-containing P=S oligonucleotides (AZPSONs) potently inhibited the HIV-l/SHIV replication and syncytium formation (ECso = 0.02-0.2 /lM) without cytotoxicity up to 100 /lM. DBM-2198, the most effective in anti-HIV-l activity among the AZPSONs, consists of random sequence and five 6¬AZNs evenly distributed in 18 nucleotides. DBM-2198 showed strong antiviral activity against, not only laboratory strains, but also primary isolates and even drug-resistant strains of HIV-I. DBM-2198 was much more effective than ddI or ddC in its anti-HIV-l activity in vitro. Particularly noteworthy is that the anti-HIV-l activity of DBM-2198 was better than that of AZT with respect to its long-lasting efficacy after a single treatment. Nevertheless, the antiviral activity of the AZPSONs was very specific to HIV-I. Poliovirus, or even simian immunodeficiency virus (SIV), was not inhibited by the AZPSONs. Taken together, our results strongly suggest that AZPSON can be used as a safe and effective AIDS-therapeutic drug against a broad spectrum of HIV -1 strains.

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Thermodynamic Investigation of the Formation of Complexes between Norfloxacin and Various Mononucleotides

  • Kwon, Yong-Jun;Lee, Hyun-Mee;Han, Sung-Wook;Lee, Dong-Jin;Cho, Tae-Sub
    • Bulletin of the Korean Chemical Society
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    • 제32권9호
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    • pp.3233-3238
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    • 2011
  • The fluorescence of norfloxacin was quenched by various nucleotides. The ratio of the fluorescence intensities in the absence and presence of nucleotide was linearly dependent on nucleotide concentration, suggesting that quenching occurred through the formation of nonfluorescent norfloxacin-nucleotide complexes. The gradient of the linear relationship represented the equilibrium constant of complex formation; it decreased with increasing temperature. The slopes of van't Hoff plots constructed from the temperature-dependent equilibrium constants were positive in all cases, indicating that complex formation was energetically favorable - i.e., exothermic, with negative Gibb's free energy. The equilibrium constant increased when triphosphate was used instead of monophosphate. It also increased when the oxygen at the $C'_2$ position of the nucleotide was removed. Both enhancements were due to entropic effects: entropy decreased when complexes with AMP or GMP formed, while it increased when norfloxacin complexed with ATP, GTP, dAMP and dGMP.

Molecular Analysis of Korean Isolate of Barley mild mosaic virus (Iks Isolate)

  • Choi, Min-Kyung;Kamala-Kannan, Seralathan;Oh, Byung-Taek;Park, Jong-Chul;Lee, Gun-Woong;Lee, Kui-Jae;Park, Yool-Jin
    • The Plant Pathology Journal
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    • 제25권2호
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    • pp.157-164
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    • 2009
  • The complete nucleotide sequences of both RNA of an isolated Barley mild mosaic virus (BaMMV) from Iksan, Korea, have been determined. RNA1 was 7273 nucleotides long and encodes for a polyprotein of 2261 amino acids, which contains the eight putative functional proteins. RNA2 was 3520 nucleotides long and encodes for a polyprotein of 894 amino acids, which contains two functional proteins. Results of multiple sequence alignment showed 92.9% similarity with Na1 isolate, followed by Sil, UK(F), Asl1, Remis M and UK(M) isolates, respectively. Comparison of the BaMMV-Iks polyproteins with the corresponding proteins of BaMMV-Na1 isolates showed 95% amino acid sequence identity. The phylogenetic analysis revealed that Iks isolate was closely related to Na1 strain and have a common origin.

Quality Attributes of Fat-free Sausage Made of Chicken Breast and Liquid Egg White

  • Lee, Hyun Jung;Jo, Cheorun;Nam, Ki Chang;Lee, Kyung Haeng
    • 한국식품영양학회지
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    • 제29권4호
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    • pp.449-455
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    • 2016
  • We developed a type of sausage made of chicken breast and liquid egg whites for consumers interested in weight management. To determine the quality of the product, its chemical characteristics, fatty acid composition, free amino acid contents, and nucleotides contents were evaluated during 4 weeks of storage. Sensory evaluation was conducted by both general consumers and body-builders. The sausage was proposed as a fat-free product as fat content was 0.12% based on the Korean Indication Standard of Animal Origin Food. Protein content was 13.42% and calorie value was 61.50 kcal/100 g of the sausage. In sensory evaluation, the mixture of chicken breast and egg whites stuffed into the same casing had an adverse effect on taste, color, texture and overall acceptance while the product that contained egg white stuffed separately into the outer casing enclosing the chicken breast (double layer) improved these attributes. The developed double-layer sausage can last for at least 4 weeks of storage without quality deterioration of flavor-related compounds, such as fatty acids and nucleotides.

$^{32}$P-Postlabelling 방법의 응용 : Azo색소 및 Flavonoid화합물에 의해 유도되는 DNA Adduct의 겸출에 관한 연구 (Application of the $^{32}$P-Postlabelling Technique : A Study on Detection of DNA Adduct Induced by Azo Dyes rind Flavonoid Compounds)

  • 김재현;박창원;박정식;홍연탁;김효정;이주한;이헌수;이동권
    • Biomolecules & Therapeutics
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    • 제1권1호
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    • pp.58-64
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    • 1993
  • DNA addicts induced by putative chemical related to carcinogenesis were detected and determined by $^{32}$P-Postlabelling assay after exposure of 4 compounds comprising two auto dyes (amaranth, new coccine) and two flavonoid compounds (rutin, quercetin) to ICR mouse. DNA was isolated from mouse liver and digested enzymatically to deoxyribonucleoside 3'-monophosphate. The postincubation of DNA digests with nuclease Pl before $^{32}$P-labelling enhanced the technique's sensitivity. Nuclease Pl cleaves deoxyribonucleoside 3'-mono-phosphates of normal nucleotides to deoxyrihonucleosides which do not serve as substrates for polynucleotide kinase, while most of addicts were found to be totally or partially resistant to the 3'-dephosphorylating action of nuclease Pl. The adducted deoxyribonucleoside 3'-monophosphate was converted to 5'-$^{32}$P-labelled deoxynucleoside 3',5'-bisphosphate by T4 polynucleotide kinase. The nucleotides were separated by anion-exchange thin layer chromatography(TLC) on polyethyleneimine cellulose by 4-dimensional or 2-dimensional TLC then detected by autoradiography. The results show that DNA addicts were detected in liver DNA of ICR mouse after administration of amaranth and quercetin by 2-dimensional and/or 4-dimensional TLC.

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