![]() ![]() Restriction enzymes recognize a specific sequence of nucleotides and produce a double-stranded cut in the DNA. Recognition site Ī palindromic recognition site reads the same on the reverse strand as it does on the forward strand when both are read in the same orientation In addition, there is mounting evidence that restriction endonucleases evolved as a selfish genetic element. ![]() Restriction enzymes likely evolved from a common ancestor and became widespread via horizontal gene transfer. The discovery of restriction enzymes allows DNA to be manipulated, leading to the development of recombinant DNA technology that has many applications, for example, allowing the large scale production of proteins such as human insulin used by diabetic patients. For their work in the discovery and characterization of restriction enzymes, the 1978 Nobel Prize for Physiology or Medicine was awarded to Werner Arber, Daniel Nathans, and Hamilton O. Later, Daniel Nathans and Kathleen Danna showed that cleavage of simian virus 40 (SV40) DNA by restriction enzymes yields specific fragments that can be separated using polyacrylamide gel electrophoresis, thus showing that restriction enzymes can also be used for mapping DNA. Restriction enzymes of this type are more useful for laboratory work as they cleave DNA at the site of their recognition sequence and are the most commonly used as a molecular biology tool. ![]() Smith, Thomas Kelly and Kent Wilcox isolated and characterized the first type II restriction enzyme, HindII, from the bacterium Haemophilus influenzae. The restriction enzymes studied by Arber and Meselson were type I restriction enzymes, which cleave DNA randomly away from the recognition site. In the 1960s, it was shown in work done in the laboratories of Werner Arber and Matthew Meselson that the restriction is caused by an enzymatic cleavage of the phage DNA, and the enzyme involved was therefore termed a restriction enzyme. If a phage becomes established in one strain, the ability of that phage to grow also becomes restricted in other strains. coli K, is known as the restricting host and appears to have the ability to reduce the biological activity of the phage λ. coli K, its yields can drop significantly, by as much as 3-5 orders of magnitude. coli C, when grown in another strain, for example E. It was found that, for a bacteriophage λ that can grow well in one strain of Escherichia coli, for example E. The phenomenon was first identified in work done in the laboratories of Salvador Luria, Jean Weigle and Giuseppe Bertani in the early 1950s. The term restriction enzyme originated from the studies of phage λ, a virus that infects bacteria, and the phenomenon of host-controlled restriction and modification of such bacterial phage or bacteriophage. These enzymes are routinely used for DNA modification in laboratories, and they are a vital tool in molecular cloning. Over 3,000 of these have been studied in detail, and more than 800 of these are available commercially. More than 3,600 restriction endonucleases are known which represent over 250 different specificities. Together, these two processes form the restriction modification system. Inside a prokaryote, the restriction enzymes selectively cut up foreign DNA in a process called restriction digestion meanwhile, host DNA is protected by a modification enzyme (a methyltransferase) that modifies the prokaryotic DNA and blocks cleavage. These enzymes are found in bacteria and archaea and provide a defense mechanism against invading viruses. To cut DNA, all restriction enzymes make two incisions, once through each sugar-phosphate backbone (i.e. Restriction enzymes are commonly classified into five types, which differ in their structure and whether they cut their DNA substrate at their recognition site, or if the recognition and cleavage sites are separate from one another. Restriction enzymes are one class of the broader endonuclease group of enzymes. A restriction enzyme, restriction endonuclease, REase, ENase or restrictase is an enzyme that cleaves DNA into fragments at or near specific recognition sites within molecules known as restriction sites. ![]()
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