Titlebook: DNA Topoisomerase Protocols; Volume I: DNA Topolo Mary-Ann Bjornsti,Neil Osheroff Book 1999 Humana Press 1999

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Sedimentation Analysis of Bacterial Nucleoid Structure,). Experiments of this type have contributed to the conclusions that (1) supercoiling is controlled in part by regulated expression of the gyrase and topoisomerase I genes, and (2) the overall level of supercoiling responds to growth environment (reviewed in [.]).

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Overexpression and Purification of ,DNA Topoisomerase III,es, using topo I preparations purified from cells containing the gene encoding topo I (.) on a multicopy plasmid showed that topo I was capable of resolving plasmid DNA replication intermediates in vitro (.). It was not until the purification and characterization of topo III that it was realized tha

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Resolution of DNA Molecules by One-Dimensional Agarose-Gel Electrophoresis,NA in agarose is also affected by the shape of the DNA, the pore size of the matrix (agarose concentration), temperature, the ionic strength of the electrophoresis buffer, the applied voltage/field strength, and the presence of intercalators (reviewed in .,.).

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Purification of Supercoiled Plasmid DNA,ils must be relaxed in order for transcription and replication to continue unaffected, as outlined in preceding chapters. The DNA topoisomerases play the central role in relaxing this supercoiling (.-.).

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Isolation of Kinetoplast DNA,NA has also proven a fascinating experimental subject in its own right. Studies of the unique structure, mechanism of replication, and genetic function of kDNA have led to numerous interesting discoveries (...-. for reviews).

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Isolation of Knotted DNA from Coliphage P4, topoisomerases. This chapter describes the isolation of knotted DNA from P4 phage. In the case of wild-type P4, such DNA molecules are double-stranded, hydrogen-bonded, knotted circles of 11,624 nucleotides.

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Overexpression and Purification of Bacterial Topoisomerase IV,ts prepared from strains overproducing the . and . gene products were mixed together. ParC and ParE were subsequently purified (.,.), and it was demonstrated that they formed a heterotetramer (.) with ATP-dependent, type II topoisomerase activity (.,.).