A protein factor which reduces the negative supercoiling requirement in the Mu DNA strand transfer reaction is Escherichia coli integration host factor.
作者： M G Surette , G Chaconas
DOI: 10.1016/S0021-9258(19)81717-9
关键词: DNA 、 Biophysics 、 Escherichia coli 、 Molecular biology 、 Host factor 、 DNA supercoil 、 Plasmid 、 Enterobacteriaceae 、 Biology 、 Transcription (biology) 、 Gel electrophoresis
摘要: We have examined the supercoiling requirement for in vitro Mu DNA strand transfer reaction and found that optimal efficiency requires a high level ( σ ¯ = −0.06) of donor plasmid superhelicity. At vivo levels −0.025) does not occur. Using an unreactive with near physiological supercoiling, we identified Escherichia coli protein factor which has novel property reducing by 40%. This protein, named relief was purified to homogeneity be identical integration host (IHF), known induce site specific bends DNA. The dramatic reduction promoted about 1.5 IHF dimers/donor substrate molecule. these low IHF, HU also reduced; synergistic effect two proteins resulted greater than 10-fold stimulation under appropriate conditions. Furthermore, at concentrations could completely eliminated from reaction.
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