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DNA-gyrase-catalyzed supercoiling. Replicating DNA generates âpositiveâ supercoils, depicted at the bottom of the diagram, as a result of rapid rotation of the DNA at the replication fork. DNA gyrase can nick and close phosphodiester bonds,
Restriction Enzymes Cut DNA Molecules at Specific Sequences. To clone specific DNA fragments in a plasmid vector, as just described, or in other vectors discussed in later sections, the fragments must be produced and then inserted into the vector DNA.As noted in the introduction, restriction enzymes and DNA ligases are utilized to produce such recombinant DNA molecules.hen such a vector is treated with a restriction enzyme that recognizes a recognition sequence in the polylinker, it is cut at that sequence, generating sticky ends. In the presence of DNA ligase, DNA fragments produced with the same restriction enzyme will be inserted into the plasmid (Figure 7-8b).