Restriction endonucleases are enzymes that bind to a specific double-stranded DNA sequence and catalyze hydrolysis of phosphodiester bonds in both DNA strands, within or near the specific sequence. Their biological function is as part of bacterial restriction-modification systems that consist of the nuclease and a DNA methyltransferase that modifies the bacterial DNA in the same sequence as recognized by the nuclease. There are four types of restriction endonucleases (types I, II, III, and IV). Of these, the type II enzymes are most useful for recombinant DNA applications, and several thousand are known. Type II restriction endonucleases that recognize and cleave DNA sequences that are 4, 5, 6, or 8 base pairs in length are commercially available. These enzyme catalyze an SN2-like nucleophilic attack by water on the phosphodiester bond. They are assisted by Mg2+ions bound in the active site. There is little amino acid sequence similarity among these enzymes, but many have in...
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