• Title/Summary/Keyword: P-gal

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Peroxiredoxin I participates in the protection of reactive oxygen species-mediated cellular senescence

  • Park, Young-Ho;Kim, Hyun-Sun;Lee, Jong-Hee;Cho, Seon-A;Kim, Jin-Man;Oh, Goo Taeg;Kang, Sang Won;Kim, Sun-Uk;Yu, Dae-Yeul
    • BMB Reports
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    • v.50 no.10
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    • pp.528-533
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    • 2017
  • Peroxiredoxin I (Prx I) plays an important role as a reactive oxygen species (ROS) scavenger in protecting and maintaining cellular homeostasis; however, the underlying mechanisms are not well understood. Here, we identified a critical role of Prx I in protecting cells against ROS-mediated cellular senescence by suppression of $p16^{INK4a}$ expression. Compared to wild-type mouse embryonic fibroblasts (WT-MEFs), Prx $I^{-/-}$ MEFs exhibited senescence-associated phenotypes. Moreover, the aged Prx $I^{-/-}$ mice showed an increased number of cells with senescence associated-${\beta}$-galactosidase (SA-${\beta}$-gal) activity in a variety of tissues. Increased ROS levels and SA-${\beta}$-gal activity, and reduction of chemical antioxidant in Prx $I^{-/-}$ MEF further supported an essential role of Prx I peroxidase activity in cellular senescence that is mediated by oxidative stress. The up-regulation of $p16^{INK4a}$ expression in Prx $I^{-/-}$ and suppression by overexpression of Prx I indicate that Prx I possibly modulate cellular senescence through $ROS/p16^{INK4a}$ pathway.

Production and Characterization of ${\beta}$-galactosidase from Bacillus licheniformis Isolated from Doenjang (된장에서 분리된 Bacillus licheniformis의 ${\beta}$-galactosidase 생산성과 효소특성)

  • Jin, Hyun Kyung;Yoon, Ki-Hong
    • Microbiology and Biotechnology Letters
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    • v.42 no.4
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    • pp.339-346
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    • 2014
  • A bacterial strain was isolated from homemade doenjang (Korean fermented soybean paste) as a producer of the extracellular ${\beta}$-galactosidase, capable of hydrolyzing lactose to liberate galactose and glucose residues. The isolate YB-1414 has been identified as Bacillus licheniformis on the basis of its 16S rDNA sequence, morphology and biochemical properties. The production of ${\beta}$-galactosidase by B. licheniformis YB-1414 reached maximum levels of 6.2 U/ml in culture medium containing wheat bran (1%) and yeast extract (2.5%) as carbon and nitrogen sources, respectively. Particularly, the insoluble fraction was more effective for ${\beta}$-galactosidase production than the soluble extract of wheat bran. The enzyme exhibited maximum activity for hydrolysis of para-nitrophenyl-${\beta}$-D-galactopyranoside (pNP-${\beta}Gal$) under reaction conditions of pH 6.0 and $55-60^{\circ}C$. Its hydrolyzing activity for pNP-${\beta}Gal$ was drastically decreased by the addition of low concentrations of galactose, but only slightly decreased by glucose, with 85% of maximal activity in the presence of 400 mM glucose.

Production and Purification of Human Lipocortin-I Secreted by Recombinant Saccharomyces cerevisiae (재조합 Saccharomyces cerevisiae로부터 인체 리포코틴-I의 분비 생산 및 정제)

  • 김병문;정봉현
    • KSBB Journal
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    • v.10 no.3
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    • pp.343-348
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    • 1995
  • Human lipocorin-I(LCI) is a calcium ion-dependent and phospholipid-binding protein which exhibits an anti-inflammatory activity by inhibiting phospholipase A2 activity. In this study, the LCI gene containing its own terminator region was joined to GAL10 promoter-ppL (prepro-leader sequence of mating factor a). An ATG start codon of LCI gene was placed at downstream with KR endoprotease recognition site(Lys-Arg) of ppL. Recombinant S. cerevisiae harboring the LCI expression/secretion vector, pYGLPT5, was aerobicall grown on a liquid YPDG medium al $30^{\circ}C$ for 72hys. The whole cell and culture supernatant were separated after centrifugation, and the expressed LCI was analyzed by SDS-PAGE and western blotting methods. A majority fraction of the expressed LCI was found to be accumulated in the intracellular fraction, resulting in very low secretion efficiency of about 7.4%. About $500mg/\ell$ of LCI was extracellularly produced by the fed-batch culture employing the controlledfeeding of glucose and galactose. The secreted LCI was purified by ultrafiltration and hydroxylapatite column chromatography, and a purity of more than 99% was obtained.

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TIMP-2 Gene Transfer Via Adenovirus Inhibits the Invasion of Lung Cancer Cell (TIMP-2 유전자 재조합 아데노바이러스의 폐암세포 침윤 억제 효과)

  • Oh, Yeon-Mok;Lee, Jae-Ho;Yoo, Chul-Gyu;Chung, Hee-Soon;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo;Lee, Choon-Taek
    • Tuberculosis and Respiratory Diseases
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    • v.49 no.2
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    • pp.189-197
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    • 2000
  • Background : Tissue inhibitor of metalloproteinase is a natural inhibitor that counteracts pro teolytic enzymes essential to the invasion of cancer cell. Whether or not TIMP-2 gene transfer via adenovirus could inhibit the invasion of lung cancer cell iη vitro was evaluated for the future purpose of gene therapy against lung cancer. Methods : Recombinant adenovirus-TIMP-2(Ad-TIMP-2) was generated by homologous recombination after pACCMV-TIMP-2 and pJM17 were cotransfected into 293 cell by standard calcium phosphate coprecipitate method. Calu-6, one of the most invasive lung cancer cells, was transduced with Ad-TIMP-2 or Ad-$\beta$gal. Anchorage-independent growth and invasiveness were assessed by soft agar clonogenicity assay and invasion assay using two-chamber, well divided by matrigel. Results : Ad-TIMP-2 transduced calu-6 cells produced biologically active TIMP-2 more than 50 times more than parental calu-6. TIMP-2 gene transfer did not suppress the in vitro tumorigenicity. However, two chamber well assay revealed that Ad-TIMP-2 transduction reduced the invasiveness of calu-6 efficiently (12% compared with parental cell) even at low 10moi. Conclusion : Even though TIMP-2 gene transfer did not inhibit in vitro tumorigenicity, it did inhibit invasion of lung cancer cell in vitro. The inhibition of invasion by Ad-TIMP-2 may be a useful strategy for the treatment of lung cancer.

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COHOMOLOGY GROUPS OF CIRCULAR UNITS

  • Kim, Jae-Moon;Oh, Seung-Ik
    • Journal of the Korean Mathematical Society
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    • v.38 no.3
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    • pp.623-631
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    • 2001
  • Let $\kappa$ be a real abelian field of conductor f and $\kappa$(sub)$\infty$ = ∪(sub)n$\geq$0$\kappa$(sub)n be its Z(sub)p-extension for an odd prime p such that płf$\phi$(f). he aim of this paper is ot compute the cohomology groups of circular units. For m>n$\geq$0, let G(sub)m,n be the Galois group Gal($\kappa$(sub)m/$\kappa$(sub)n) and C(sub)m be the group of circular units of $\kappa$(sub)m. Let l be the number of prime ideals of $\kappa$ above p. Then, for mm>n$\geq$0, we have (1) C(sub)m(sup)G(sub)m,n = C(sub)n, (2) H(sup)i(G(sub)m,n, C(sub)m) = (Z/p(sup)m-n Z)(sup)l-1 if i is even, (3) H(sup)i(G(sub)m,n, C(sub)m) = (Z/P(sup)m-n Z)(sup l) if i is odd (※Equations, See Full-text).

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Changes in Physicochemical Quality during the Seasoned and Fermented Alaska Pollack Roe with Vacuum Fermentation (조미 명란(明卵)의 진공발효 및 저장중 이화학적 품질 변화)

  • 김정욱;이성갑
    • The Korean Journal of Food And Nutrition
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    • v.16 no.4
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    • pp.353-358
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    • 2003
  • Low salt-fermented product of alaska pollack roe were prepared tentatively and also discussed the retarding effect of fermentation period by vacuum. The results were as follows. The moisture content, pH and salinity decreased slightly, but the contents of VBN and total amino acids increased gradually during fermentation regardless of ripening types. pH and salinity showed little difference with ripening types, but moisture content was a little lower and the contents of VBN was some higher in non vacuum than those in vacuum, whereas total amino acid contents were greater in vacuum. In addition, total viable cell count were similar to trend of gentle decrement after increment to some degree but were higher in non vacuum than those in vacuum. Judging from the results of physicochemical analysis, it was showed that fermentation delayed in vacuum therefore it was considered that vacuum fermentation can be effective on shelf-life extension of jeot-gal.

Construction of a Lactococcal Shuttle/Expression Vector Containing a $\beta$-Galactosidase Gene as a Screening Marker (선별마커로써 $\beta$-Galactosidase 유전자를 포함한 Lactococcus용 셔틀/발현 벡터 제조)

  • Han Tae Un;Jeong Do-Won;Cho San Ho;Lee Jong-Hoon;Chung Dae Kyun;Lee Hyong Joo
    • Microbiology and Biotechnology Letters
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    • v.33 no.4
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    • pp.241-247
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    • 2005
  • A new lactococcal shuttle/expression vector for lactococci, pWgal13T, was constructed using a $\beta$-galactosi-dase gene (lacZ) from Lacfococcus lactis ssp. lactis ATCC 7962 as a screening marker. The pWgal 13T was introduced into Escherichia coli DH5a and L. lactis MG1363, and was easily detected by the formation of blue colonies on a medium containing X-gal without any false transformants. Also, the quantitatively lacZ activity of pWgal13T was measured in L. lactis ssp. cremoris MG1363, and was found to be four times higher than that of L. lactis ssp. lactis ATCC7962 grown on a medium containing glucose, which shows that the lacZ gene of pWgal13T can be used for the efficient screening of L. lactis on general media. The pWgal13T was equipped with a lactococcal replicon of pWV01 from L. lactis Wg2, the new promoter P13C from L. lactis ssp. cremoris LM0230, multiple cloning sites, and a terminator for the expression of a relevant gene. The vee-tor pWgal13T was used for the expression of the EGFP gene in E. coli and L. lactis. These results show that the lactococcal expression/shuttle vector constructed in the present study can be used for the production of foreign proteins in E. coli and L. lactis.

NMR and Fluorescence Studies of DNA Binding Domain of INI1/hSNF5

  • Lee, Dongju;Moon, Sunjin;Yun, Jihye;Kim, Eunhee;Cheong, Chaejoon;Lee, Weontae
    • Bulletin of the Korean Chemical Society
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    • v.35 no.9
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    • pp.2753-2757
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    • 2014
  • INtegrase Interactor 1 protein (INI1/hSNF5) or BRG1-associated factor 47 (BAF47) is a SWI/SNF-related matrix associated actin dependent regulator of chromatin subfamily B member. DNA binding domain of INI1/hSNF5 is cloned into E.coli expression vectors, pET32a and purified as a monomer using size exclusion chromatography. NMR data show that $INI1^{DBD}$ has folded state with high population of ${\alpha}$-helices. By fluorescence-quenching experiments, binding affinities between $INI1^{DBD}$ and two double stranded DNA fragments were determined as $29.9{\pm}2.6{\mu}M$ (GAL4_1) and $258.7{\pm}5.8$ (GAL4_2) ${\mu}M$, respectively. Our data revealed that DNA binding domain of INI1/hSNF5 binds to transcriptional DNA sequences, and it could play an important role as a transcriptional regulator.

Effect of IPTG Induction on Production of ${\beta}$-Galactosidase-PreS2 Fusion Protein in Recombinant Escherichia coli

  • Nam, Soo-Man;Park, Young-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.1 no.4
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    • pp.274-280
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    • 1991
  • Effects of IPTG induction on cell growth and production of ${\beta}$-galactosidase-preS2 fusion protein (${\beta}$gal-preS2) were studied in a defined medium using a recombinant Escherichia coli JM109/pCMHB30. IPTG was added (0.2 mM) to induce the cloned-gene expression in the early-, mid-, and late-log growth phases. The most serious decreases in growth rate and plasmid stability were observed for the induction in the early-log growth phase. The expression level of ${\beta}$gal-preS2 attained by the induction in the mid-log phase was about 0.51 mg fusion protein/mg total cellular protein, which was 2- and 5-fold improvement over the levels obtained with the inductions in the early- and late-log phases. Formation of acidic byproducts including acetate and pyruvate showed different profiles during the fermentation period for each cases of induction; pyruvate was the major byproduct for the induction in the early-log phase while acetate production became more significant for the cases of inductions in the mid- and late-log phases.

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Patrinia monandra (Valerianaceae): an unrecorded species in Korea (긴뚝갈(마타리과): 국내 미기록종)

  • Kim, Jin-Seok;Lee, Byung-Chun;Chung, Jea-Min;Pak, Jae-Hong
    • Korean Journal of Plant Taxonomy
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    • v.34 no.2
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    • pp.167-172
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    • 2004
  • We report an unrecorded species of Patrinia in Korea, P. monandra C. B. Clarke. This species was found from Neasosa, Buan-gun, Prov. Jeollabuk-do, Sanghwangbong and Bogildo, Wando-gun, Prov. Jeollanam-do in the southwest of Korea. We named 'Gin-dook-gal' as a common name of this species. It is easily distinguishable from Patrlnia villosa (Thunb.) Juss. by the characteristics such as lemon yellow corolla, one (sometimes three) stamen(s) and fruit fused with large bracteole-wing orbiculate.