50 research outputs found

    Dam methylase from Escherichia coli: kinetic studies using modified DNA oligomers: hemimethylated substrates.

    No full text
    We have measured steady-state kinetics of the N6-adenine methyltransferase Dam Mtase using as substrates non-selfcomplementary tetradecamer duplexs (d[GCCGGATCTAGACG]-d[CGTCTAGATCC-GGC]) containing the hemimethylated GATC target sequence in one or the other strand and modifications in the GATC target sequence of the complementary strands. Modifications included substitution of guanine by hypoxanthine (I), thymine by uracil (U) or 5-ethyl-uracil (E) and adenine by 2,6-diamino-purine (D). Thermodynamic parameters were obtained from the concentration dependence of the melting temperature (Tm) of the duplexes. Large differences in DNA methylation of duplexes containing single dI for dG substitution of the Dam recognition site were observed compared with the canonical substrate, if the substitution involved the top strand (on the G.C rich side). Substitution in either strand by uracil (dU) or 5-ethyluracil (dE) resulted in small perturbation of the methylation patterns. When 2,6-diamino-purine (dD) replaced the adenine to be methylated, small, but significant methylation was observed. The kinetic parameters of the methylation reaction were compared with the thermodynamic free energies and significant correlation was observed

    Hyperfine interaction with the 229^{229}Th nucleus and its low-lying isomeric state

    No full text
    The thorium nucleus with mass number A=229A=229 has attracted much interest because its extremely low lying first excited isomeric state at about 88eV opens the possibility for the development of a nuclear clock. However, neither the exact energy of this nuclear isomer nor properties, such as nuclear magnetic dipole and electric quadrupole moment are known to a high precision so far. The latter can be determined by investigating the hyperfine structure of thorium atoms or ions. Due to its electronic structure and the long lifetime of the nuclear isomeric state, Th2+^{2+} is especially suitable for such kind of studies. In this letter we present a combined experimental and theoretical investigation of the hyperfine structure of the 229^{229}Th2+^{2+} ion in the nuclear ground and isomeric state. A very good agreement between theory and experiment is found for the nuclear ground state. Moreover, we use our calculations to confirm the recently presented experimental value for the nuclear magnetic dipole moment of the thorium nuclear isomer, which was in contradiction to previous theoretical studies
    corecore