• Title/Summary/Keyword: cos-cleavage

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Isolation and Characterization of the Smallest Bacteriophage P4 Derivatives Packaged into P4-Size Head in Bacteriophage P2-P4 System

  • Kim, Kyoung-Jin;Song, Jae-Ho
    • Journal of Microbiology
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    • v.44 no.5
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    • pp.530-536
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    • 2006
  • Bacteriophage P4, a satellite phage of coliphage P2, is a very useful experimental tool for the study of viral capsid assembly and cos-cleavage. For an in vitro cos-cleavage reaction study of the P2-P4 system, new shortened and selectable markers containing P4 derivative plasm ids were designed as a substrate molecules. They were constructed by swapping the non-essential segment of P4 DNA for either the kanamycin resistance (kmr) gene or the ampicillin resistance (apr) gene. The size of the genomes of the resulting markers were 82% (P4 ash8 delRI:: kmr) and 79% (P4 ash8 delRI:: apr) of the wild type P4 genome. To determine the lower limit of genome size that could be packaged into the small P4-size bead, these shortened P4 plasmids were converted to phage particles with infection of the helper phage P2. The conversion of plasmid P4 derivatives to bacteriophage particles was verified by the heat stability test and the burst size determination experiment. CsCl buoyant equilibrium density gradient experiments confirmed not only the genome size of the viable phage form of shortened P4 derivatives, but also their packaging into the small P4-size head. P4 ash8 delRI:: apr turned out to be the smallest P4 genome that can be packaged into P4-sized head.

Isolation and Identification of the Bacteriophage P4 ost2, Suppressing sir Mutations of Bacteriophage P2 (박테리오파아지 P2의 sir 변이를 억제하는 새로운 박테리오파아지 P4 유도체인 P4 ost2의 분리와 동정)

  • Kim, Gyeong-Jin
    • Korean Journal of Microbiology
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    • v.39 no.4
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    • pp.277-282
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    • 2003
  • Bacteriophage P4 ost2 which is the P4 mutant suppressing sir mutations of Bacteriophage P2, was isolated as a plaque-former by plating P4 ash8 sid 71 kmr intS on the lawn of P2 sir3 lysogen. P4 ost2 turned out to be the P4 mutant suppressing sir mutations of P2 in one-step growth experiments.

A Protein Tyrosine Phosphatase Inhibitor, Pervanadate, Inhibits Angiotensin II-Induced β-Arrestin Cleavage

  • Jang, Sei-Heon;Hwang, Si Ae;Kim, Mijin;Yun, Sung-Hae;Kim, Moon-Sook;Karnik, Sadashiva S.;Lee, ChangWoo
    • Molecules and Cells
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    • v.28 no.1
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    • pp.25-30
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    • 2009
  • ${\beta}$-Arrestins turn off G protein-mediated signals and initiate distinct G protein-independent signaling pathways. We previously demonstrated that angiotensin $AT_1$ receptorbound ${\beta}$-arrestin 1 is cleaved after $Phe^{388}$ upon angiotensin II stimulation. The mechanism and signaling pathway of angiotensin II-induced ${\beta}$-arrestin cleavage remain largely unknown. Here, we show that protein Tyr phosphatase activity is involved in the regulation of ${\beta}$-arrestin 1 cleavage. Tagging of green fluorescent protein (GFP) either to the N-terminus or C-terminus of ${\beta}$-arrestin 1 induced conformational changes and the cleavage of ${\beta}$-arrestin 1 without angiotensin $AT_1$ receptor activation. Orthovanadate and molybdate, inhibitors of protein Tyr phosphatase, attenuated the cleavage of C-terminal GFP-tagged ${\beta}$-arrestin 1 in vitro. The inhibitory effects of okadaic acid and pyrophosphate, which are inhibitors of protein Ser/Thr phosphatase, were less than those of protein Tyr phosphatase inhibitors. Cell-permeable pervanadate inhibited angiotensin II-induced cleavage of ${\beta}$-arrestin 1 in COS-1 cells. Our findings suggest that Tyr phosphorylation signaling is involved in the regulation of angiotensin II-induced ${\beta}$-arrestin cleavage.

Isolation, Characterization, and Application of Chitosan-Degrading Fungus from Soil

  • Wei, Xinlin;Chen, Wei;Xiao, Ming;Xiao, Jianbo;Wang, Yuanfeng
    • Journal of Microbiology and Biotechnology
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    • v.20 no.7
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    • pp.1114-1120
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    • 2010
  • A chitosan-degrading fungus, BSF114, was isolated from soil. The culture preparation showed strong chitosanolytic enzyme activity at an optimum pH of 4.0 and optimum temperature of $60^{\circ}C$ after 36-40 h fermentation. The rapid decrease in the viscosity of the chitosan solution early in the reaction suggested an endo-type cleavage of the polymeric chitosan chains. To identify the isolated fungus, molecular biological and morphological methods were used. The fungal internal transcribed spacer (ITS) region 1 was amplified, sequenced, and then compared with related sequences in the GenBank database using BLAST. The phylogenetic relationships were then analyzed, and the results showed that the fungus belongs to Aspergillus fumigatus. Morphological observations were also used to confirm the above conclusion. The chitooligosaccharides (COS) obtained through hydrolyzing the colloidal chitosan showed that A. fumigatus BSF114 is suitable for degrading chitosan and producing chitooligosaccharides on a large scale. High concentrations of the COS (1,000 and 500 ${\mu}g/ml$) significantly proliferated mice marrow cells.

Evaluation for Rock Cleavage Using Distribution of Microcrack Spacings (V) (미세균열의 간격 분포를 이용한 결의 평가(V))

  • Park, Deok-Won
    • The Journal of the Petrological Society of Korea
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    • v.26 no.3
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    • pp.297-309
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    • 2017
  • Jurassic granite from Geochang was analysed with respect to the characteristics of the rock cleavage. The comprehensive evaluation for rock cleavages was performed through the combination of the 16 parameters derived from the enlarged photomicrographs of the thin section and the spacing-cumulative frequency diagrams. The results of analysis for the representative values of these spacing parameters with respect to the rock cleavage are summarized as follows. First, the above parameters can be classified into group I (spacing frequency (N), total spacing ($1m{\geq}$), constant (a), exponent (${\lambda}$), slope of exponential straight line (${\theta}$), length of line (oa') and trigonometric ratios ($sin{\theta}$, $tan{\theta}$) and group II (mean spacing (Sm), difference value between mean spacing and median spacing (Sm-Sme), density (${\rho}$), lengths of lines (oa and aa'), area of a right-angled triangle (${\Delta}oaa^{\prime}$) and trigonometric ratio($cos{\theta}$). The values of the 8 parameters belonging to group I show an order of H(hardway, H1+H2)