When restriction enzymes were first isolated in 1970 there were not many applications to which they could be put to use. they are now an important tool in genetic engineering. briefly list the human needs and demands that have driven the development and use of restriction enzymes in genetic engineering:

The diagnosis of genetic diseases related to changes in the DNA sequence, gene cloning, discover certain gene alleles.

Explanation:

A restriction enzyme is one that can recognize a characteristic sequence of nucleotides within a DNA molecule and cut the DNA at that particular point, called a restriction site or target, or at a site not too far away. Restriction sites have between four and six base pairs, with which they are recognized. The use of these enzymes as a biotechnological tool arose from the need to cut the long strands of genomic DNA to be manipulated and analyzed into smaller fragments. The use of restriction enzymes and other enzymes that modify nucleic acids, such as ligase, allowed the development of recombinant DNA methodology.

The mechanism of cutting DNA is through the breakage of two phosphodiester bonds in the double strand, resulting in two ends of DNA. These can be either blunt (when the broken links match) or cohesive/staggered.

Some of the fields in which restriction enzymes have been most implicated has been:

The diagnosis of genetic diseases related to changes in the DNA sequence: Whether they are point mutations, insertions or deletions of fragments. The procedure for such a diagnosis would consist of amplifying by PCR a specific region of the chromosome where we know that this genetic change should or should not be and adding the restriction enzyme or enzymes so that the relevant cuts are made. After making these cuts, we will run our samples in an electrophoresis gel and see the differences in size that exist. If the mutation generates a deletion, when running the electrophoresis we will see a smaller fragment than we would expect from a healthy patient, so that we can diagnose it. If the mutation, on the contrary, generates a point mutation, we will be able to take advantage of it to use an enzyme at that point and thus, if there is no such mutation, that cut will not be produced and we will obtain a larger fragment in the electrophoresis gel. So, when this enzyme is applied to the gene of a healthy and a sick person, different amounts of fragments should be observed for each case in an electrophoresis.They can also be used during gene cloning: To clone a gene, it must be cut out of the DNA using restriction enzymes. This fragment is then inserted into a cloning vector, which is a plasmid that increases the number of copies. Once this is obtained, restriction enzymes are used to cut the gene from the clone vector and put it into an expression vector so that the gene is expressed and the protein of interest is produced.Be used to discover certain gene alleles: Similar to the first point, different amounts of fragments should be observed for each case in an electrophoresis if there are different gene alleles

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