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- 2025年07月08日
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文献和实验Overexpression and Purification of Bacterial DNA Gyrase
DNA gyrase is the bacterial type II topoisomerase that can introduce negative supercoils into DNA using the free energy of ATP hydrolysis (1 ,2 ). The enzyme from Escherichia coli consists of two proteins, A and B (termed GyrA and GyrB
Isolation of Covalent Enzyme-DNA Complexes
linkage with the 5′-ends of the cleaved strands of DNA (2 ). In contrast, cleavage of DNA by the type I–3′-enzymes (i.e., eukaryotic topoisomerase I, DNA topoisomerase V, and vaccinia, variola, and Shope fibroma viral topoisomerases) results
E.Z.N.A.® Endo-free Plasmid Mini Kit I Spin Protocol
as above. This step ensures that residual protein contamination is removed and must be included for downstream application requiring high quality DNA. 12. Discard the flow-through liquid and wash the column by adding 700 u DNA Wash Buffer diluted with ethanol
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