Application
In molecular biology research, restriction endonucleases are frequently employed for the following purposes:
- Genetic engineering: Restriction endonucleases are most frequently used as a technique in genetic engineering. The host organism’s genome can be modified and interesting sequences can be introduced thanks to the endonuclease activity. The host then produces the desired gene product as a result. This idea has numerous biotechnological applications, including the creation of antibiotics, antibodies, enzymes, and several secondary metabolites.
- Restriction enzymes are used in DNA mapping, sometimes referred to as restriction mapping, to gather structural data on the DNA fragment. This method produces DNA fragments of varied sizes by digesting the DNA with a sequence of restriction enzymes. By using agarose gel electrophoresis to separate the resulting fragments, the separation between the restriction enzyme sites can be calculated. This can be used to ascertain a DNA fragment’s structure.
- Gene sequencing entails digesting a big DNA molecule with restriction enzymes and then running the resultant pieces through a DNA sequencer to determine the nucleotide sequence.
- Studies on gene expression, mutations, and population polymorphisms are some of the other uses for restriction endonucleases.
Restriction Enzymes
Restriction enzyme is a bacterial protein that cleaves DNA at particular locations, these sites are called restricted sites. The restriction enzymes guard against bacteriophages in living bacteria. They identify the bacteriophage and cleave it at its restriction sites, destroying its DNA. Important genetic engineering tools include restriction enzymes. They may be separated from bacteria and applied in research facilities. The recognition sequences, or short and distinct nucleotide sequences, are recognized by restriction enzymes in DNA. When a DNA sequence is recognized by the restriction enzyme, it hydrolyzes the bond between neighboring nucleotides and cleaves the DNA molecule. The bacteria use the enzyme methylases to add the methyl group at the adenine or cytosine bases within the recognition sequence, preventing the DNA sequences from disintegrating.