• Title/Summary/Keyword: ultrasoft X-ray

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The Effects on Escherichia coli and Plasmid DNA Using Ultrasoft X-ray (Ultrasoft X-ray의 Escherichia coli균과 plasmid DNA에 대한 영향)

  • ;;;;;Seiya Chiba;Atsuo Kimura
    • KSBB Journal
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    • v.15 no.1
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    • pp.84-87
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    • 2000
  • We studied the effect of ultrasoft X-ray obtained from the Pohang Light Source (PLS), on the mutation of E. coli and the damage of plasmid. After irradiation, the supercoiled plasmid DNA converted to the relaxed-form, and then to the linear-form. We transformed the irradiated plasmid DNA and isolated $\beta$-galactosidase mutants. We also isolated $\beta$-galactosidase mutants from the directly irradiated cells. There were preferred mutational sites on DNA induced by ultrasoft X-ray.

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Direct Evidence for the Radioprotective Effect of Various Carbohydrates on Plasmid DNA and Escherichia coli Cells

  • Ryu, Hwa-Ja;Yi, Kyung-Eun;Kim, Do-Won;Jung, Yun-Dae;Chang, Suk-Sang;Seo, Eun-Seong;Lee, Ki-Young;Marceau-Day, M.L.;Kim, Do-Man
    • Journal of Microbiology and Biotechnology
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    • v.12 no.4
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    • pp.598-602
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    • 2002
  • Damage to cells exposed to radiation is primarily attributed to direct effects on the structure of cellular DNA. Radiation-induced damage of pBluescript SK plasmid DNA and Escherichia coli $DH5\alpha$ were examined in the presence of various branched oligosaccharides, polysaccharides, and/or 8-MOP (8-methoxypsoralen). Branched oligosaccharides efficiently protected DNA and cells exposed to ultrasoft X-ray and UV irradiation. In the presence of 0.2% (w/v) branched oligosaccharides and polysaccharides, DNA can be protected from damage due to W and ultrasoft X-ray by a factor of 1.3-2.1 fo1d and 3.2-8.3 fold, respectively. The protective effect of cells exposed to UV or ultrasoft X-ray was also observed by branched oligosaccharides. The combination of MOP, a photoreagent, with carbohydrates increased the protective effects for DNA and cells, compared with that of a single use of MOP or carbohydrate alone.

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

  • Gang, Hui-Gyeong;Kim, Do-Man;Jang, Seok-Sang
    • 한국생물공학회:학술대회논문집
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    • 2003.04a
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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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